Milking stools in the collections of the National Cultural History Museum: an investigation of typology. In this study the physical or other characteristics that might typify the traditional milking stool is investigated. This was done by studying a comparable group of eighteen examples from the Anthropology and Cultural History collections of the National Cultural History Museum in Pretoria. Six of the objects in the group studied had already been associated with the Afrikaans catchword ‘melkstoel’, hence the primary sources that informed the research included the museological data from the existing documentation of the Museum. The investigation confirmed that corresponding physical attributes, especially that pertaining to construction, could be identified in the group studied. It was also found that the objects had seen multiple, but indeterminable, functional applications. Since fifteen of the eighteen examples studied were essentially anonymous, as no documented provenance existed, their origin, history and meaning could not be established. The research was therefore extended to secondary sources, being textual descriptions and images, from which associations and correlations between the group studied and milking stools, especially from Western Europe and the English-speaking world, could be affirmed. Moreover, such links were also found to exist with other stools and certain vernacular chair typologies of shared approach, especially as to their construction. Findings identified certain typological features of the milking stool, but also a symbolic and metaphorical association. Multiple functions of and applications for the milking stool could be ascertained. AFRIKAANS : In hierdie studie word ondersoek ingestel of enige eienskappe, fisies of andersins, die tradisionele melkstoeltjie tipeer. Dit is onderneem aan die hand van ‘n studiegroep van agtien soortgelyke voorwerpe uit die Antropologie-versameling en die Kultuurhistoriese versameling van die Nasionale Kultuurhistoriese Museum, Pretoria. Dié primêre bronne word in die ondersoek ondersteun deur museale bronne ontsluit uit die Museum se dokumentasiesentrum, waar ses van die voorwerpe in die studiegroep reeds assosieer is met die trefwoord ‘melkstoel’. Die ondersoek het bevestig dat sekere fisiese kenmerke van die studiegroep, veral in hul konstruksie, dui op groepsverband. Daar is ook bevind dat die voorwerpe vir meervoudige, maar onbepaalde, funksies gebruik is.Van die agtien voorwerpe in die studiegroep is vyftien van anonieme herkoms, sodat hul oorsprong, geskiedenis en betekenis nie met sekerheid vasgestel kon word nie. Die navorsing is daarom uitgebrei na tekstuele beskrywings en beelde, as sekondêre bronne, om hierdie en ander kenmerke te ondersoek. Hieruit is die studiegroep in verband gebring met melkstoeltjies uit veral Wes-Europa en die Engelssprekende wêreld, asook ander stoeltjies en volkstoele wat soortgelyke eienskappe en konstruksiekenmerke het. Bevindings dui op bepaalde tipologiese kenmerke van ‘n melkstoeltjie, maar ook ‘n wyer, assosiatiewe begrip wat manifesteer in beeldspraak en ‘n metaforiese konteks. Meervoudige gebruike en aanwendings van die melkstoeltjie is nagespeur. Copyright / Dissertation (MInt)--University of Pretoria, 2010. / Architecture / unrestricted

O método de diagnóstico coproparasitológico de concentração em formolacetato de etila foi empregado para a quantificação de ovos de helmintos. O método quantitativo proposto foi padronizado utilizando-se o Sistema Comercial Coprotest® e amostras fecais contendo diferentes cargas de ovos de Ascaris lumbricoides. O \"Coprotest quantitativo\" foi aplicado em estudo populacional numa área de baixa endemicidade para parasitas diagnosticáveis através das fezes, e os resultados analisados comparativamente ao método de Kato-Katz. A metodologia proposta foi ainda comparada a outros métodos quantitativos, utilizando-se amostras fecais, preparadas em laboratório, com cargas decrescentes de ovos de A. lumbricoides, Trichuris trichiura e Schistosoma mansoni. Discutem-se as vantagens de se empregar um método capaz de detectar o maior número de espécies, tanto de helmintos quanto de protozoários, . e que permita, concomitantemente, estimar a intensidade das infecções por geohelmintos e S. mansoni nas populações. O \"Coprotest quantitativo\" mostrou ser de aplicação viável em trabalhos de campo, fornecendo resultados comparáveis aos outros métodos quantitativos já descritos na literatura. / Abstracts not available

L’action en urbanisme demeure un champ de recherche riche au regard de sa complexité. Par projet, elle est pratiquée selon une anticipation opératoire partiellement déterminée et repose sur des croyances. Le projet d’urbanisme se définit par l’articulation de plusieurs ambiguïtés notamment entre l’intention directrice de l’action et la confrontation de la réalisation à la réalité. Cette recherche participe à l’intelligibilité de la pratique du projet d’urbanisme, pratique de la complexité. Comme opérateur du projet d’urbanisme, les valeurs constituent l’objet de cette recherche. Nous proposons d’éclairer la pratique du projet par l’analyse de la mobilisation des valeurs dans, pour et par le projet. L’intrigue de cette recherche est la relation entre la mobilisation des valeurs dans le projet d’urbanisme et celui-Ci. Les hypothèses de ce travail reposent sur l’interaction entre la dynamique de cette mobilisation et la dynamique du projet. L’étude du projet Bossut à Cergy-Pontoise, pensé depuis plus de quatre décennies, et l’analyse de la mobilisation des valeurs dans ce projet, celles de Centralité, d’Identité et de Lien, ont révélé que les valeurs, si elles portent le projet d’urbanisme en retour sont portées par lui. Représentée par la figure du Tripode-Culbuto, la dynamique, à la fois stable et instable, de la mobilisation des valeurs dans, pour et par le projet, permet de gérer et de pratiquer une partie de la complexité du projet d’urbanisme. / Action in urban planning remains a field of research rich in terms of its complexity. Each project is performed based on an anticipated, partially determined, operating process and on beliefs as opposed to evidence. An urban planning project is defined as the interlink between several ambiguities, notably between the intended action of the planning and the reality of the situation. This research contributes to a better understanding of the practical considerations of an urban project, an exercise of complexity. The values as operators in urban planning constitute the subject of this research. We propose to provide substantial insights into the practicalities of urban planning by analyzing the mobilization of these values, in, for and by the project. The intrigue of this research is the relationship between the mobilization of these values in urban planning and the project itself. The hypotheses of this work rely on the interaction between the dynamic of this mobilization and the dynamic of the project. A study of the Bossut project in Cergy-Pontoise, which has been in the planning stages for four decades, and an analysis of the mobilization of the values herein (centrality, identity and connection), revealed that they did indeed support the project and, in return, were supported by it. Represented by the figure three-Legged stool / “Culbuto”, the dynamic of the mobilization of values in, for, and by the project, at times stable and unstable, allows managing and putting into practice some of the complexity of urban planning.

Humanpatogena stammar av bakterien Yersinia enterocolitica kan orsaka akut gastroenterit. För identifiering av bakterien odlas fecesprover ut på CIN-agar. På senare år har en kromogen agarplatta framtagits som differentierar mellan patogena och apatogena stammar av Y. enterocolitica. Syftet med studien är att jämföra och utvärdera två CIN-agar, med agarbaser och supplement från två olika företag (Liofilchem och Oxoid), och CHROMagar Y. enterocolitica (CHROMagar). Odling av fecesprover samt seriespädning av sex Y. enterocolitica stammar och en Y. pseudotuberculosis utfördes. Vid utodlade fecesprover jämfördes växt och hämning av övriga bakterier. Vid seriespädning räknades antal kolonier på plattorna för respektive spädning, samt utseende av kolonier på plattor bedömdes. Resultatet tyder på att skillnad av hämningseffekt av Y. enterocolitica och utseende på kolonierna finns mellan de två CIN-agarplattorna. Oxoid’s CIN-agar erhöll större kolonier, lägre hämningseffekt av Y. enterocolitica och detektionsgräns än Liofilchem’s CIN-agar. På CHROMagar-plattan växte de patogena stammarna med bleklila kolonier och de apatogena stammarna med blåa kolonier. Hämningseffekt av Y. enterocolitica hos CHROMagar-plattan är densamma som Oxoid’s CIN-agar. Slutsatsen är således att Oxoid’s CIN-agar och CHROMagar har samma hämningseffekt av Y. enterocolitica men CHROMagar differentierar mellan patogena och apatogena stammar. Liofilchem’s CIN-agar har högre hämningseffekt än CHROMagar och Oxoid’s CIN-agar. / Pathogenic strains of Yersinia enterocolitica can cause acute gastroenteritis in humans. To identify the bacterium, cultivation of stool samples on CIN-agar are performed. A chromogenic medium has been developed that differentiate between pathogenic and nonpathogenic strains of Y. enterocolitica. The purpose is to compare and evaluate two CIN-agar, with agar bases and supplements from two companies (Liofilchem and Oxoid), and CHROMagar Y. enterocolitica (CHROMagar). Growth of stool samples and serial dilutions of six Y. enterocolitica strains and one strain of Y. pseudotuberculosis were performed. Comparisons of the growth and inhibition of other bacteria were done for the stool samples. Colonies for each dilution were counted and appearance of the colonies was evaluated. The result indicates that a difference in inhibitory effect on Y. enterocolitica and appearance of colonies exist between the two CIN-agar. All strains grew with larger colonies on Oxoid CIN-agar than on Liofilchem’s. Oxoid CIN-agar and CHROMagar have a lower inhibitory effect on Y. enterocolitica than Liofilchem’s. On CHROMagar, the pathogenic strains grew with mauve colonies, whilst the nonpathogenic strains grew with blue colonies. Thus, the conclusion is that CHROMagar and Oxoid CIN-agar have less inhibitory effect on Y. enterocolitica than Liofilchem’s. CHROMagar can differentiate between pathogenic and nonpathogenic strains.

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No. of bitstreams: 2 Dissertação - Carolina Silva Petenusse - 2017.pdf: 5090665 bytes, checksum: 4e27e8d1ee68369b7e5c2ea4625bf67f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-02-15 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Helminthic zoonosis constitutes part of the public and collective health problems due to the ever-closer approximation between humans and animals. There is a shortage of information about the contamination, prevention and care forms to be taken to avoid these conditions which makes difficult to reduce it on the population. Therefore, it is essential to research about the life cycle of parasites and its human-animal relation. Among the transmission forms, soil stands out, since parasitized animals defecate on the soil and disseminate eggs, what makes possible the infection of others. Including the places that the animals usually visit, there are parks and groves of the cities. Therefore, the aim of this research was to analyze on the collected dogs and cat’s stool samples on these places in Goiânia, Goiás, the existence of helminths that can cause diseases in humans, as well as to evaluate the perception of the people who visit these places about the zoonosis and the hygiene care of their pets. It was also aimed to present a literature review about the helminths found during the research, that are transmitted by cats and dog’s feces in urban areas. Nine parks/groves were visited, one per city region, to the collection of the feces and parasitological tool analysis and a quiz with 30 alternative and discursive questions was applied to 164 interviewees which affirmed to have a shortage knowledge about zoonosis. Out of 116 analyzed samples, the coproparasitological test presented the presence of Ancylostoma spp. in 11.21% and Toxocara spp. in 2.5% of them. Regarding the questionnaire, most of people were dog owners (68.9%) and 82.84% of them reportedly gave anthelmintics to their animals. When asked about zoonosis, only 4.88% of people knew its meaning. Despite most people do not have any knowledge about zoonosis, the results may be associated, due to the better care of people with their pets, but this still was not enough to avoid the transmission of these helminths. Actions and measures should be taken, such as creating a practice of monitoring a potential zoonotic parasite in public places by public health agencies, for the purpose of aiming at the prevention against the parasitosis and few spending with treatment. / As zoonoses helmínticas constituem parte dos problemas em saúde pública e coletiva devido a aproximação cada vez mais estreita entre humanos e animais. Há escassez de informações acerca das formas de contaminação, prevenção e cuidados a serem tomados para evitar essas afecções, o que dificulta sua redução na população. Dessa forma, é fundamental que sejam realizados estudos sobre os ciclos de vida de parasitos e sua relação homem-animal. Dentre as formas de transmissão desses, o solo se destaca, uma vez que animais parasitados defecam no solo e disseminam ovos, o que possibilita a infecção alheia. Entre os locais em que estes costumam frequentar em comum se encontram os parques e os bosques dos municípios. Logo, o objetivo deste trabalho foi analisar nas amostras fecais de cães e gatos colhidas nesses locais de Goiânia, Goiás, a existência de helmintos que possam causar doenças em humanos, bem como, avaliar a percepção das pessoas que visitam estes locais acerca das zoonoses e cuidados de higiene com seus animais de estimação. Também visou-se apresentar uma revisão de literatura a respeito das helmintoses encontradas durante a pesquisa, que são veiculadas por meio das fezes de cães e gatos em ambiente urbano. Foram visitados nove parques/bosques municipais, um por região da cidade, para colheita das fezes e análises coproparasitológicas e aplicação de um questionário a 164 entrevistados, que confirmaram o conhecimento escasso sobre zoonose. Nos exames coproparasitológicos detectaram-se 11,21% de ovos de Ancylostoma spp. e 2,5% de Toxocara spp. em 116 amostras fecais. Quanto ao inquérito, a maioria das pessoas possuíam cão em casa (68,9%) e 82,84% destas afirmaram administrar anti-helmínticos. Sobre as zoonoses, apenas 4,88% das pessoas sabiam seu significado. Os resultados podem estar associados entre si, devido ao melhor cuidado das pessoas com seus animais de estimação, porém ainda não é o suficiente para evitar a transmissão dessas helmintoses. Ações e medidas profiláticas devem ser tomadas, como criar uma prática habitual de monitoramento de parasitos com potencial zoonótico em locais públicos por meio de órgãos públicos da saúde, com a finalidade de visar a prevenção contra essas parasitoses e menos gastos com tratamentos.

O método de diagnóstico coproparasitológico de concentração em formolacetato de etila foi empregado para a quantificação de ovos de helmintos. O método quantitativo proposto foi padronizado utilizando-se o Sistema Comercial Coprotest® e amostras fecais contendo diferentes cargas de ovos de Ascaris lumbricoides. O \"Coprotest quantitativo\" foi aplicado em estudo populacional numa área de baixa endemicidade para parasitas diagnosticáveis através das fezes, e os resultados analisados comparativamente ao método de Kato-Katz. A metodologia proposta foi ainda comparada a outros métodos quantitativos, utilizando-se amostras fecais, preparadas em laboratório, com cargas decrescentes de ovos de A. lumbricoides, Trichuris trichiura e Schistosoma mansoni. Discutem-se as vantagens de se empregar um método capaz de detectar o maior número de espécies, tanto de helmintos quanto de protozoários, . e que permita, concomitantemente, estimar a intensidade das infecções por geohelmintos e S. mansoni nas populações. O \"Coprotest quantitativo\" mostrou ser de aplicação viável em trabalhos de campo, fornecendo resultados comparáveis aos outros métodos quantitativos já descritos na literatura. / Abstracts not available

Les méthodes pour le diagnostic des infections à mycobactéries sont restées pratiquement inchangées pendant des décennies et n'auraient probablement pas progressé sans la réémergence inattendue de la tuberculose (TB) au cours des vingt dernières années du 20ème siècle. Le diagnostic de laboratoire de la tuberculose pulmonaire repose principalement sur la détection des organismes du complexe Mycobacterium tuberculosis (MTC) dans les prélèvements respiratoires par leur isolement et identification ou par les méthodes moléculaires. Pour les patients qui n’expectorent pas, des échantillons du tractus respiratoire peuvent être obtenus par des procédures invasives. En nous basant sur la survie connue de MTC dans le liquide gastrique, nous avons considéré que les MTC devraient également être détectables dans les selles. Nous avons recherché des MTC en parallèle dans des échantillons de tractus respiratoire et dans les selles récoltés des mêmes patients. Mycobacterium tuberculosis a été détecté après décontamination à la chlorhexidine dans des cultures sur des milieux à base d'œuf et par l'examen direct microscopique après coloration de Ziehl-Neelsen. Après la mise au point d’une technique originale d’extraction de l’ADN total à partir des selles, nous avons utilisé la détection de M. tuberculosis par PCR en temps réel en duplex utilisant le gène IS6110 comme cible moléculaire et un contrôle interne de présence d’inhibiteurs de la PCR. Ces travaux mettent en évidence l’intérêt et la sensibilité de l’analyse des selles pour le diagnostic de la tuberculose pulmonaire. Cette technique est aujourd’hui utilisée dans notre laboratoire en routine et est en cours d’évaluation par d’autres équipes.Actuellement, l'identification des mycobactéries basée sur l’analyse des caractéristiques biochimiques (test à l’uréase, test de la nitrate réductase, test de la résistance à la pyrazinamide (pza), test de la sensibilité, test de la résistance à la thiophen-2-carboxylic acid hydrazide (TCH) etc …) est le plus souvent remplacée par des méthodes de biologie moléculaire dont l’amplification par PCR d'un gène spécifique. Ces méthodes sont laborieuses et peuvent exiger un degré d'expertise élevé. Afin de simplifier l'identification précise des isolats de mycobactéries en laboratoire de routine, après avoir vérifié l’inactivation des MTC par la chaleur et l’éthanol pour pouvoir travailler hors du laboratoire P3, nous avons mis au point les conditions pour l'utilisation de la spectrométrie de masse « matrix-assisted laser desorption and ionisation-time-of-flight» (MALDI-TOF-MS) en association avec un logiciel bioinformatique spécifique, comme outil d’identification et de différenciation entre les membres du complexe MTC, du complexe Mycobacterium avium et les autres mycobactéries non-tuberculeuses. Nous avons ensuite appliqué cette technique afin d’identifier les mycobactéries à partir du milieu liquide Middlebrook 7H10 utilisé dans une étuve automatisée. L’identification des mycobactéries isolées dans notre laboratoire, est maintenant réalisée en routine par MALDI-TOF-MS. L’ensemble de ces travaux contribue à améliorer le diagnostic de routine de la tuberculose pulmonaire et les techniques mises au point sont utilisées en routine dans notre laboratoire, en particulier dans le cadre d’un « kit tuberculose ». / The diagnosis of mycobacterial infections remained practically unchanged during numerous decades and would not probably have progressed of the whole without the unexpected re-emergence of the tuberculosis (TB) during the last twenty years of the 20th century.The diagnosis of laboratory of pulmonary tuberculosis is mainly based on the detection of microorganisms of the Mycobacterium tuberculosis complex (MTC) in the respiratory specimens. For the patients who do not expectorate, samples of the respiratory tract can be only obtained according to invasive procedures. Based on the survival known for MTC organisms in the gastric liquid, we considered that MTC organisms would be detectable also in samples of stools. We compared the presence of bodies MTC in samples of respiratory tract and the specimens of stools collected from the same patients. MTC was detected in cultures on egg-based media after appropriate decontamination using the chlorhexidine and by the microscopic examination after Ziehl-Neelsen staining. After the set on of an original protocol of the total DNA extraction from stools, we were able to use by the detection with the real-time PCR of IS6110 using internal controls as PCR inhibitor control. This protocol illustrate the utility of stool analysis for the diagnosis of pulmonary tuberculosis, this technique is actually used in routine in our laboratory, it is also under investigation by other research teams.Phenotypic identification of mycobacteria based on the analysis of biochemical pattern (urase test, nitrate reductase test, resistance to pyrazinamide (pza) test, susceptibility to thiophen-2-carboxylic acid hydrazide (TCH) test, etc …) is more often replaced by molecular methods using DNA, including the development by PCR of a specific gene. These methods are generally laborious and can require a considerable degree of expertise. To simplify the identification specifies isolates of mycobacteria routinely in the laboratory, we estimated the use of the mass spectrometry "matrix-assisted laser desorption and ionization-time-of-flight" (MALDI-TOF MS) in association with a specific bioIT software, was capable of identifying and of distinguishing between the members of Mycobacterium tuberculosis complex, Mycobacterium avium complex and the non- tuberculosis mycobacteria. We verified the MTC inactivation by heating or by ethanol allowing the analysis of the inactivated MTC out of the P3 laboratory; we studied the cost-effectiveness of this method from the blood solid media by extrapolating this approach in the emergent countries. We then applied the same method to identify mycobacteria from Middlebrook 7H10 liquid media used in an automated oven. Identification of cultured Mycobacteria is now done in routine in our laboratory by MALDI-TOF-MS. These data contribute in improving the routine diagnosis of pulmonary tuberculosis and this protocol is then used routinely in our laboratory, particularly with the “kit de tuberculose” we set on.

The intestinal microbiota is composed of up to 100 trillion microorganisms of which bacteria are overwhelming majority. The microbiota affects the development of the immune system, defence against pathogens, host nutrition, vitamin synthesis or fat storage and its composition is changing throughout life. Some studies point to an association between microbiota composition and the development of inflammatory bowel disease. One of the treatment options is anti-TNFα antibodies therapy, which uptake or antagonize the TNFα cytokine that otherwise mediates inflammation in the intestinal mucosa. The aim of the thesis was to examine how this treatment affects the composition of the intestinal bacteriome in paediatric patients with Crohn's disease, and to find specific bacterial taxa, whose abundance changes during the treatment. By inclusion of patients with juvenile idiopathic arthritis, also treated with anti-TNFα, the study aims to discern specific effects of therapeutically induced intestinal restitution (observable in patients with Crohn's disease) from general effects of anti-TNFα therapy. Stool samples from healthy children were used to determine "healthy" bacteriome. The composition of the bacteriome was studied by profiling the variable region of the V4 gene of 16S rDNA from patients stool samples...

This dissertation describes the synthesis, antimicrobial properties, and catalytic activity of a variety of eta5-ligand rhodium and iridium complexes. Cp*RM(beta-diketonato)Cl (Cp*R = R-substituted tetramethylcyclopentadienyl ligand) complexes were found to have selective activity against Mycobacterium smegmatis, with activity highly dependent upon the substituents on the Cp*R ligand as well as on the beta-diketonato ligand. These complexes were synthesized in good yield from the reaction of the chloro bridged dimers ([Cp*RMCl2]2) with the desired beta-diketonato ligand under basic conditions. All complexes were fully characterized by 1H and 13C NMR. Twenty single crystal X-ray structures were solved. The success of these syntheses led to investigation of another beta-diketonato ligand: 1,1,1,5,5,5-hexafluoroacetylacetonate (hfac). Though many metal complexes of this ligand are known, reaction with [Cp*MCl2]2 did not yield the desired Cp*M(hfac)Cl complexes. Instead, a variety of products were obtained, three of which were characterized crystallographically. The most interesting structure featured a non-coordinating trifluoroacetate (TFA) anion and a [Cp*Ir]3Na1O4 cubane structure, which is an unprecedented and highly unusual arrangement for iridium. Attempts to synthesize this cluster rationally through reactions of [Cp*IrCl2]2 with TFA yielded instead a chloro bridged [Cp*IrCl(TFA)] dimer. Reaction of [Cp*MCl2]2 with 1,1,1-trifluoroacetylacetonate (tfac) yielded the expected Cp*M(tfac)Cl complex, indicating that the problem lies with using hfac as a ligand for Cp*M(III) complexes. Finally, the indenyl effect was investigated for the oxidative annulation of 2-phenylimidazole with 1-phenyl-1-propyne catalyzed by a series of methyl-substituted [(indenyl)RhCl2] dimers. [(Ind*)RhCl2]2 was found to have significantly greater activity than [Cp*RhCl2]2 (100% vs. 51%). Two plausible catalytic cycles were proposed, one of which invokes a ring slip transition state. Though it is unclear if the "indenyl effect" is responsible for this differing activity, it is certainly apparent that using an indenyl ligand has a notable effect in this catalytic reaction. Cyclometalation was also investigated stoichiometrically for 2-phenyl-1H-imidazole and 1-phenylpyrazole and found to proceed readily for [(Ind*)RhCl2]2. Additionally, the crystallographic structure of a Rh+ /Rh– ionic pair was solved. Ionic pairs such as this are rarely found in the literature. / Doctor of Philosophy / This dissertation deals with the uses of a series of unusual compounds containing the metals rhodium and iridium. Though these are rare and expensive metals, the uses and benefits described in this dissertation far outweigh the costs. Overall, the compounds described in this dissertation are colorfully characterized as “piano stool” compounds because of their overall shape and appearance. The metal, either rhodium or iridium, occupies a central point in the complex. On top of the metal is a “flat” organic group that gives the appearance of the seat of the piano stool. Below the metal, there are three other groups that look like the legs of the piano stool. By appropriate choice of the metals and the surrounding groups, special properties can be designed into these “piano stool” complexes. Chapter 2 describes the synthesis of a series of complexes where the “flat” group is a variant of a five-membered carbon ring compound known as cyclopentadienyl, the metal is rhodium or iridium, and two of the three legs come from a family of compounds known as acetylacetonates (acac). This series of piano stool compounds display antimicrobial activity against a class of pathogens known as mycobacteria, an example of which causes the disease tuberculosis. Changing the cyclopentadienyl group and the acac group allows for this antimicrobial activity to be tuned. In the following chapter, attempts to make the same type of compound described in the paragraph above with fluorine-substituted acacs gave some very unexpected results. The most surprising result was a very unusual cube-shaped structure containing 3 iridium atoms, 1 sodium atom, and 4 oxygen atoms, which is an unprecedented arrangement for iridium. Finally, there is a specific example of a flat group for the piano stool known as indenyl. Indenyl is intriguing because it can change shape from a flat group to a bent group. In doing this, it provides more space around the metal for other molecules to bind. The result of this work shows that piano stool compounds created with this indenyl group are more active and selective for carrying out a catalytic reaction to make new ring systems that could have potential use in the synthesis of new flavorings, fragrances, and even pharmaceuticals.

Les Archaea methanogènes sont des organismes environnementaux ayant été également détectés dans certaines flores associées aux muqueuses des mammifères. Chez l’homme ces microorganismes ont été associés avec les muqueuses intestinale, vaginale et orale. Ces organismes sont des procaryotes anaérobies stricts et leurs conditions de culture restent fastidieuses et très mal connues. En effet, uniquement trois Archaea methanogènes ont été cultivées à partir de prélèvements humains, Methanobrevibater smithii et Methanosphaera stadtmanae à partir des selles puis Methanobrevibater oralis à partir de la plaque dentaire. Récemment l’ADN d’autres Archaea methanogènes et d’Archaea non-methanogènes a été détecté dans des selles humaines, y compris des séquences indiquant la présence d’espèces appartenant à un nouvel ordre de méthanogènes n’ayant aucun représentant cultivé. La connaissance actuelle sur la diversité de ces methanogènes chez l’homme et sur leurs effets potentiels sur la santé humaine est en grande partie basée sur les techniques de détection de l’ADN par PCR et métagénomique. Ces techniques fondées sur la détection de l’ADN ribosomal 16S et du gène mcrA codant la sous-unité alpha du methyl-coenzyme M reductase, une enzyme clé dans le processus de méthanogenèse, ont montré dans un premier temps que M. smithii était détecté chez moins de 50% des individus et M. stadtmanae chez 0-20 % seulement. Ces résultats étaient contradictoires avec le rôle de la méthanogenèse dans l’élimination des acides et d’autres produits du processus digestion, et nous avons émis l’hypothèse que ces résultats pouvaient ne pas refléter la quantité réelle des méthanogènes dans le tube digestif humain, suggérant la mise au point de nouvelles méthodes de détection moléculaire et de culture adaptées aux caractéristiques de ces organismes fastidieux. Dans ce travail, nous nous sommes fixés comme premier objectif de mettre au point une méthode moléculaire permettant de détecter M. smithii chez tous les individus testés et nous avons mis au point un protocole d’extraction et de détection d’ADN d’Archaea à partir des selles en se basant sur les génomes séquencés de M. smithii et M. stadtmanae. Ce protocole nous a permis de détecter M. smithii chez 95,5% des individus et M. stadtmanae chez 29,4% des individus. En ce basant sur ce protocole et moyennant une approche moléculaire basée sur une PCR universelle de l’ADN ribosomal 16S des méthanogènes, le séquençage et le clonage, nous avons également détecté chez 4% de la population, une séquence correspondant à un phylotype (FJ823135) ayant déjà été rapporté comme représentant un nouvel ordre de méthanogènes. A partir de là, nous avons choisi un prélèvement de selle susceptible de contenir le plus fort ratio de FJ823135/ M. smithii et nous avons réussi à isoler et à cultiver une nouvelle Archaea que nous avons nommé Methanomassiliicoccus luminyensis, premier représentant cultivé d’un nouvel ordre de méthanogènes et la quatrième Archaea cultivée chez l’homme. M. luminyensis et M. stadtmanae présentent des métabolismes similaires en réduisant le méthanol en méthane en utilisant l’hydrogène comme donneur d’électrons, cette observation nous a incité à tester l’addition de tungstate de sélénium, requis pour la croissance de M. luminyensis, dans une culture M. stadtmanae, et nous avons observé une accélération de la vitesse de croissance de M. stadtmanae par un facteur 3. Nous avons ensuite étudié la sensibilité des méthanogènes isolés chez l’homme aux antibiotiques et établi qu’ils sont seulement sensibles à des molécules efficaces contre les bactéries et les eucaryotes, ceci étant en accord avec leur position phylogénétique en tant qu’un des quatre domaines de la vie. [...] / Methanogenic Archaea are environmental organisms which have also been associated to mammals mucosa. In humans these microorganisms have been detected in the vaginal, intestinal and oral mucosa. These organisms are strict anaerobes and their culture conditions remains fastidious and poorly known. In fact only three methanogens have been isolated from human samples, both Methanobrevibater smithii and Methanosphaera stadtmanae from stool and Methanobrevibater oralis from dental plaque. Current knowledge on the diversity of methanogens in humans and their potential effects on human health were largely based on DNA detection methods as PCR and metagenomics. These techniques based on 16S rDNA and mcrA gene (encoding the alpha subunit of methyl coenzyme-M-reductase, a key enzyme in methanogenesis process) detection, showed that M. smithii was the most present in man and that the presence of M. stadtmanae was transient. Recently, the DNA of other methanogenic and non- methanogenic Archaea, has been detected in human feces, including sequences indicating the presence of non-cultured species belonging to potential new order of methanogens with no cultured representative. However, these studies detected M. smithii with variable prevalence in less than half of the tested individuals and no M. stadtmanae; such results does not confirm the paramount role of methanogenesis in preventing the accumulation of acids and other reaction end products during the digestion process, and can not reflect the actual amount of these two methanogens in the human digestive tract because of their specific association with the intestinal mucosa. Therefore, these studies pointed that the diversity of methanogens in humans has been underestimated suggesting the development of new molecular detection methods and cultural approaches adapted these fastidious organisms. In this work, we preset as first criteria, the detection of M. smithii in all tested individuals, therefore we developed an improved protocol for archaeal DNA extraction and detection from stool based on sequenced genomes of M. smithii and M. stadtmanae, this protocol allowed us to detect the first one DNA in 95.5% tested individuals and the second in a prevalence of 29.4%. Based on this protocol and through molecular approach based on universal amplification of methanogenic 16S rDNA, sequencing and cloning, we detected in 4% of the tested population, a sequence corresponding to a new phylotype (FJ823135) that has been previously reported and proposed as a representative of a new order of methanogens. From there, we chose one stool specimen susceptible to contain the highest amount of FJ823135 and successfully isolated Methanomassiliicoccus luminyensis B10T clone, the first cultured representative of a new order of methanogens and the fourth Archaea cultured in humans.This archaeon exhibited a similar type of metabolism to that of M. stadtmanae by oxidizing H2 and reducing methanol to methane but require tungstate-selenite, an element essential for its growth, this fact prompted us testing tungstate-selenite addition on M. stadtmanae growth and establishing that it was strongly stimulatory with a growth rate three times faster. We have thereafter studied the sensitivity of methanogens isolated from humans to antibiotics and established that they are susceptible only to molecules also effective against both Bacteria and Eucarya, in agreement with their phylogenetic location as a unique domain of life. The aim of the latter part of this work was to test the effectiveness of MALDI-TOF mass spectrometry identification of environmental and host-associated Archaea. The obtained data indicated that that MALDI-TOF-MS protein profiling is an efficient first-line step for the rapid phenotypic identification of cultured Archaea organisms including host-associated ones. [...]