Direct three-dimensional myocardial strain tensor quantification and tracking using zHARP.

Images of myocardial strain can be used to diagnose heart disease, plan and monitor treatment, and to learn about cardiac structure and function. Three-dimensional (3D) strain is typically quantified using many magnetic resonance (MR) images obtained in two or three orthogonal planes. Problems with this approach include long scan times, image misregistration, and through-plane motion. This article presents a novel method for calculating cardiac 3D strain using a stack of two or more images acquired in only one orientation. The zHARP pulse sequence encodes in-plane motion using MR tagging and out-of-plane motion using phase encoding, and has been previously shown to be capable of computing 3D displacement within a single image plane. Here, data from two adjacent image planes are combined to yield a 3D strain tensor at each pixel; stacks of zHARP images can be used to derive stacked arrays of 3D strain tensors without imaging multiple orientations and without numerical interpolation. The performance and accuracy of the method is demonstrated in vitro on a phantom and in vivo in four healthy adult human subjects.

A polymorphic microsatellite in the tumor necrosis factor alpha promoter identifies an allele unique to the NZW mouse strain.

We have amplified a (CA)n:(GT)n microsatellite from the TNF promoters of a panel of mouse strains using the polymerase chain reaction. The length of the microsatellites was polymorphic, with eight alleles observed among 15 inbred strains bearing seven distinct H-2 haplotypes, and four outbred strains. In B10 congenic strains, the TNF allele detected by microsatellite polymorphism segregated with the MHC, and in recombinant haplotypes (NOD, NZW), it segregated with H-2D. The TNF allele found in the NZW strain (H-2z) was distinct from those of all other haplotypes, consistent with the hypothesis that this strain may carry a genetic defect in TNF production.

Fluid-rock interaction during late stages of the Alpine exhumation

Résumé de la thèseLa fracturation des roches au cours de phases compressives ou extensives est un souvent évoquée pour expliquer la circulation de fluide au sein des roches cristallines. Dans le cadre de cette thèse, la circulation des fluides lors de l'exhumation tardive des Alpes a été étudiée en utilisant deux approches différentes: analyses structurales de la déformation fragile d'une part et analyses géochimiques des roches et des minéraux (isotopes stables, datations U/Pb, thermochronologie (U-Th)/He) d'autre part. Cette approche combinée a permis de mieux comprendre l'interaction existante entre les fluides métamorphiques et les fluides météoriques, ainsi que leur interaction avec les roches encaissantes. Le travail a été effectué dans la zone Pennique du Valais suisse.La première partie était focalisée sur la déformation fragile, le but étant de définir les différents types de déformations existantes et de déterminer l'âge relatif des différentes familles de failles. Dans la région d'étude, quatre domaines ont été distingués. Chacun d'eux comportent deux types de structures fragiles, certaines sont minéralisées alors que d'autre non. Au sein de chaque domaine, la direction principale des structures minéralisées correspond à l'orientation des accidents tectoniques majeurs de la région (Aosta- Ranzola Line au Sud, Rhône Line au Nord et Simplon Fault Zone à l'Est), alors que les structures non- minéralisées montrent des orientations plus variables. Ainsi, le premier type de structure est interprété comme résultant d'une dislocation tectonique alors que le deuxième type de structure résulterait d'une dislocation gravitaire locale. Il n'est néanmoins pas possible de classer chronologiquement la formation de ces deux types de structure ni d'attribuer un âge relatif aux changements d'orientation des contraintes majeures.La deuxième étude a été effectuée dans la région de la zone de faille du Simplon. Dans cette zone, la composition isotopique des minéraux ayant cristallisé à l'intérieur des fractures tardives permet de distinguer différents types de circulation de fluide. Les valeurs δ180 du quartz de la roche encaissante ainsi que ceux des veines tardives du bloque inférieur de la faille sont comparables. Ces valeurs indiquent un rééquilibrage et un tamponnage isotopique des fluides tardifs au contact de la roche encaissante lors de la fracturation de cette dernière et de la cristallisation des veines tardives. La même situation est observée dans la partie nord du bloque supérieur ainsi que dans sa partie sud. Ceci n'est néanmoins pas le cas pour la partie centrale du bloque supérieur où les valeurs isotopiques des minéraux dans les veines tardives sont approximativement 3 %o plus basses (avec des valeurs extrêmes négatifs), indiquant une contribution d'eau météorique aux fluides circulant dans les veines. Ces données suggèrent qu'une infiltration d'eau météorique a pu avoir lieu dans le bloque supérieur, où la fracturation des roches est plus intensive car le déplacement relatif le long de la faille y fut plus important, et la température maximale du métamorphisme plus basse. La troisième contribution traite de la géo-thermochronologie de la zone de contact entre la klippe de la Dent Blanche et la nappe de Tsaté. De petits zircons euhédraux ont été trouvés dans un plan de faille minéralisé (parallèle à la Faille du Rhône, voir première partie de l'étude), riche en hématite et quartz, de la zone d'étude. Les analyses U/Pb donnent des âges radiométriques autour de 270 - 280 Ma aux zircons extraits de la minéralisation ainsi que ceux extraits de la roche encaissante, ce qui correspond à l'âge de la nappe de la Dent Blanche et non celui de la nappe du Tsaté qui est elle-même classiquement interprétée comme une ophiolite Jurassique de l'Océan Liguro-Piémontais. Ces données suggèrent que les zircons contenus dans la veine ont été hérités de la roche encaissante. Les résultats (U-Th)/He indiquent un âge de refroidissement différent pour la roche encaissante (25.5 ± 2.0 Ma) que celui de la minéralisation (17.7 ±1.4 Ma). Le thermomètre isotopique quartz-hématite indique une température d'équilibre, et donc de mise en place de la minéralisation, d'environ 170 °C, température très proche de la température de -180 °C de fermeture du zircon pour le système (U-Th)/He. Ceci suggère que l'âge de refroidissement des zircons de la minéralisation correspond aussi à l'âge de formation de la faille.Thesis abstractFluid circulation in fractured rocks is a common process in geology, and it is generally the consequence of faulting and fracturing during both tectonic compression and extension. This thesis is focused on fluid circulation during late stages of the Alpine exhumation. After a structural analysis of the late brittle deformation of the studied samples, several analytical methods (stable isotope investigations, U/Pb radiometric dating, (U-Th)/He thermochronology) have been applied to understand the interaction of metamorphic and meteoric fluids with one another as well as with the host rock. This thesis is articulated around three study directions. All studies were conducted in the Penninic Zone of the Valais, Switzerland. The first study deals with late, brittle deformation and focuses on the different deformation styles and on the relative age of the different families of fractures. In order to do this, late brittle structures observed in four different domains have been subdivided as a function of the existence (or not) and type of mineralization. Comparisons between mineralized and non-mineralized strike directions for all four domains show that mineralized structures follow the strike orientation of major tectonic movements indicated in the Penninic Zone of the Valais (Aosta-Ranzola Line to the S, Rhône Line to the Ν and Simplon Fault Zone to the E), whereas non-mineralized fractures have a more variable strike orientation. This difference could be interpreted as indicative of tectonic-related faulting (mineralized structures) vs. local, collapse-related faulting (non-mineralized fractures), but it is not strong enough to indicate a relative age of the late brittle structures, and/or a change in the orientation of the strain field in post-Miocene times. The second studied area is focused on the Simplon Fault Zone (SFZ). Stable isotope analyses of minerals filling these late fractures indicate that there are two different fluid circulation systems in the footwall and hanging wall of the SFZ. In the footwall, δ180 values of quartz from both the host rock and the late veins range from +10 %o to +12 %o. This is consistent with buffering of circulating fluids by the host rock during fracturing and vein precipitation. In the hanging wall, δΙ80 values for quartz crystals from the host rock and the late veins are similar in both the northern and southern parts of the detachment that are both affected by the same degree of metamorphism (greenschist to the Ν and amphibolite to the S). This is not the case in the central part of the SFZ, where there is a jump from amphibolite facies in the footwall to greenschist facies in the hanging wall. δ,80 values for quartz from the hanging wall late veins are approximately 3.0 %o lower (down to negative values in some cases) than the values observed in the footwall These data suggest that infiltration of meteoric water may have occurred in the most fractured parts of the hanging wall, where relative displacement on the SFZ was the greatest and the peak temperature lower. In the less fractured footwall the δ180 values reflect a host rock-buffered system.The third study is focused on geo-thermochronology at the contact between the Dent Blanche nappe and the Tsaté nappe where small, euhedral zircons were found in a hematite- and quartz-rich mineralization on a late normal fault plane parallel to the Rhône Line (see first part of the study). U/Pb analysis indicates that the zircons - both in the late mineralization and in the host rock - have absolute radiometric ages clustering around 270 - 280 Ma, which is the accepted age for intrusive rocks from the Austroalpine Dent Blanche units but not for the Tsaté nappe. The latter is classically interpreted as an ophiolitic remnant of the Jurassic Liguro-Piemontais Ocean. U/Pb analyses suggest that zircons in late mineralization are all inherited from the host rock; however, results of (U-Th)/He analyses indicate that cooling ages for the host rocks are different to the cooling ages for the zircons in late mineralization. Indeed, the calculated cooling age for the Arolla gneiss is 25.5 ± 2.0 Ma, whilst the cooling age for the associated mineralized fault plane is 17.7 ±1.4 Ma. Oxygen stable isotope fractionation between quartz and hematite in the same late mineralization corresponds to temperatures of about 170 °C. The proximity of the calculated emplacement temperature for the mineralization and the lower accepted closure temperature for zircon in the (U-Th)/He system (-180 °C) imply that the age of 17.7 ± 1.4 Ma can also be interpreted as the formation age of this late brittle fault.Résumé grand publicLa circulation des fluides dans les roches fracturées est typique de nombreux processus géologiques, et très souvent est la conséquence de la fracturation des roches. Cette thèse aborde la question de la circulation des fluides pendant les dernières phases du soulèvement des Alpes. Après une analyse structurale de la fracturation directement sur le terrain, plusieurs méthodes géochimiques ont été appliquées pour comprendre l'interaction entre les différents fluides circulants, et avec leur propre roche mère. L'étude, concentrée sur trois directions principales, a été conduite dans la zone Pennique du Valais suisse. La première partie traite de la déformation cassante dans le secteur cité. L'analyse détaillée des fractures a permis de les subdiviser en structures minéralisées et non-minéralisées, sur quatre domaines différents. La comparaison entre les directions des structures minéralisées et non-minéralisées a permis de montrer que les premières suivent l'orientation des accidents tectoniques majeurs de la région, alors que les structures non- minéralisées ont une orientation plus variable. Cette différence pourrait être interprétée comme indication d'une dislocation tectonique (structures minéralisées) contre une dislocation gravitaire locale (structures non-minéralisées), mais elle n'est pas assez forte pour indiquer un âge relatif des structures tardives et/ou un changement de l'orientation des contraintes après -20 Ma vers le présent.A partir de ces observations, la deuxième étude est concentrée dans la région de la faille du Simplon. Les analyses géochimiques sur les minéraux remplissant les structures tardives indiquent qu'il y a deux différents systèmes de circulation des fluides dans les deux parties (toit et mur) de la faille. Dans le mur, les valeurs isotopiques des minéraux cristallisés à partir d'un fluide tardif sont les mêmes de ceux de la roche mère, donc il y a eu rééquilibration chimique entre fluide et roche pendant la fracturation de cette dernière et la précipitation des minéraux. Dans le toit, les valeurs isotopiques dans la roche mère et dans les minéraux des veines tardives sont comparables dans les parties Ν et S de la faille, où les roches du toit et du mur ont atteint une température maximale - pendant phase prograde de la formation des Alpes - comparable. Au contraire, dans la partie centrale, où le mur a atteint des températures maximales plus élevées par rapport au toit, les valeurs géochimiques des minéralisations tardives du toit sont parfois plus basses que les valeurs observées dans le mur. Ces données suggèrent que l'infiltration de l'eau de surface aurait pu se produire dans la partie plus fracturée du toit, où le déplacement relatif le long de la faille était majeur et les températures maximales mineures. Au contraire, les données géochimiques du mur de la partie centrale indiquent un système isotopique équilibré par la roche mère.La troisième partie de ce travail se base sur l'étude géochimique intégrée des isotopes stables d'Oxygène et radioactifs du Plomb, Uranium, Thorium et Hélium, auprès d'une faille normale minéralisée et des roches de la région à cheval entre deux nappes, la nappe de la Dent Blanche et la nappe de Tsaté. Ici, des petits zircons ont été trouvés dans la minéralisation citée, riche en hématite et quartz. L'analyse radiométrique Uranium/Plomb a montré que les zircons dans la minéralisation et dans les roches autour ont des âges comparables (autour 280 Ma). Cela signifie que les zircons dans la minéralisation tardive ont été hérités de la roche mère pendant la fracturation et la circulation des fluides tardives. De l'autre coté, les résultats des analyses Uranium-Thorium/Hélium indiquent que les âges de refroidissement pour les roches mères sont différents comparés aux âges de refroidissement pour les zircons dans la minéralisation tardive: ces derniers sont plus jeunes d'environ 8 Ma (autour 25 Ma et autour 17 Ma respectivement). Les analyses des isotopes de l'oxygène sur quartz et hématite dans la même minéralisation donnent une température de mise en place de cette dernière d'environ 170° C. La température de fermeture du système chimique des zircons dans le système (Uranium-Thorium)/Hélium est d'environ 180 °C: la proximité de ces deux températures implique que l'âge de refroidissement de la minéralisation tardive peut également être interprété comme âge de formation de la faille.

Tissue distribution of the Ankara strain of vaccinia virus (MVA) after mucosal or systemic administration.

MVA is a candidate vector for vaccination against pathogens and tumors. Little is known about its behaviour in mucosal tissues. We have investigated the fate and biosafety of MVA, when inoculated by different routes in C57BL/6 mice. Intranasal inoculation targeted the virus to the nasal associated lymphoid tissue and the lungs, whereas systemic inoculation led to distribution of MVA in almost all lymphoid organs, lungs and ovaries. Intravaginal, intrarectal and intragastric inoculations failed to induce efficient infection. After 48 h no virus was detectable any more in the organs analyzed. Upon intranasal inoculation, no inflammatory reactions were detected in the central nervous system as well as the upper and lower airways. These results show the tropism of MVA and indicate that high doses of recombinant MVA are safe when nasally administered, a vaccination route known to elicit strong cellular and humoral immune responses in the female genital tract.

Deleterious impact of a virulent bacteriophage on survival and biocontrol activity of Pseudomonas fluorescens strain CHAO in natural soil.

Many biotic and abiotic factors affect the persistence and activity of beneficial pseudomonads introduced into soil to suppress plant diseases. One such factor may be the presence of virulent bacteriophages that decimate the population of the introduced bacteria, thereby reducing their beneficial effect. We have isolated a lytic bacteriophage (phi)GP100) that specifically infects the biocontrol bacterium Pseudomonas fluorescens CHA0 and some closely related Pseudomonas strains. phiGP100 was found to be a double-stranded-DNA phage with an icosahedral head, a stubby tail, and a genome size of approximately 50 kb. Replication of phiGP100 was negatively affected at temperatures higher than 25 degrees C. phiGP100 had a negative impact on the population size and the biocontrol activity of P. fluorescens strain CHA0-Rif (a rifampicin-resistant variant of CHA0) in natural soil microcosms. In the presence of phiGP100, the population size of strain CHA0-Rif in soil and on cucumber roots was reduced more than 100-fold. As a consequence, the bacterium's capacity to protect cucumber against a root disease caused by the pathogenic oomycete Pythium ultimum was entirely abolished. In contrast, the phage affected neither root colonization and nor the disease suppressive effect of a phiDGP100-resistant variant of strain CHA0-Rif. To our knowledge, this study is the first to illustrate the potential of phages to impair biocontrol performance of beneficial bacteria released into the natural soil environment.

Contribution of the Global Regulator Gene gacA to Persistence and Dissemination of Pseudomonas fluorescens Biocontrol Strain CHA0 Introduced into Soil Microcosms.

Structural and regulatory genes involved in the synthesis of antimicrobial metabolites are essential for the biocontrol activity of fluorescent pseudomonads and, in principle, amenable to genetic engineering for strain improvement. An eventual large-scale release of such bacteria raises the question of whether such genes also contribute to the persistence and dissemination of the bacteria in soil ecosystems. Pseudomonas fluorescens wild-type strain CHA0 protects plants against a variety of fungal diseases and produces several antimicrobial metabolites. The regulatory gene gacA globally controls antibiotic production and is crucial for disease suppression in CHA0. This gene also regulates the production of extracellular protease and phospholipase. The contribution of gacA to survival and vertical translocation of CHA0 in soil microcosms of increasing complexity was studied in coinoculation experiments with the wild type and a gacA mutant which lacks antibiotics and some exoenzymes. Both strains were marked with spontaneous resistance to rifampin. In a closed system with sterile soil, strain CHA0 and the gacA mutant multiplied for several weeks, whereas these strains declined exponentially in nonsterile soil of different Swiss origins. The gacA mutant was less persistent in nonrhizosphere raw soil than was the wild type, but no competitive disadvantage when colonizing the rhizosphere and roots of wheat was found in the particular soil type and during the period studied. Vertical translocation was assessed after strains had been applied to undisturbed, long (60-cm) or short (20-cm) soil columns, both planted with wheat. A smaller number of cells of the gacA mutant than of the wild type were detected in the percolated water and in different depths of the soil column. Single-strain inoculation gave similar results in all microcosms tested. We conclude that mutation in a single regulatory gene involved in antibiotic and exoenzyme synthesis can affect the survival of P. fluorescens more profoundly in unplanted soil than in the rhizosphere.

Signal peptide and helical bundle domains of virulent canine distemper virus fusion protein restrict fusogenicity.

Persistence in canine distemper virus (CDV) infection is correlated with very limited cell-cell fusion and lack of cytolysis induced by the neurovirulent A75/17-CDV compared to that of the cytolytic Onderstepoort vaccine strain. We have previously shown that this difference was at least in part due to the amino acid sequence of the fusion (F) protein (P. Plattet, J. P. Rivals, B. Zuber, J. M. Brunner, A. Zurbriggen, and R. Wittek, Virology 337:312-326, 2005). Here, we investigated the molecular mechanisms of the neurovirulent CDV F protein underlying limited membrane fusion activity. By exchanging the signal peptide between both F CDV strains or replacing it with an exogenous signal peptide, we demonstrated that this domain controlled intracellular and consequently cell surface protein expression, thus indirectly modulating fusogenicity. In addition, by serially passaging a poorly fusogenic virus and selecting a syncytium-forming variant, we identified the mutation L372W as being responsible for this change of phenotype. Intriguingly, residue L372 potentially is located in the helical bundle domain of the F(1) subunit. We showed that this mutation drastically increased fusion activity of F proteins of both CDV strains in a signal peptide-independent manner. Due to its unique structure even among morbilliviruses, our findings with respect to the signal peptide are likely to be specifically relevant to CDV, whereas the results related to the helical bundle add new insights to our growing understanding of this class of F proteins. We conclude that different mechanisms involving multiple domains of the neurovirulent A75/17-CDV F protein act in concert to limit fusion activity, preventing lysis of infected cells, which ultimately may favor viral persistence.

Parasites and sociality in ants

SUMMARY : Parasites and sociality in ants This thesis investigates the complex relationships between sociality, defences against parasites and the regulation of social structures. We studied how fungal parasites influenced colony organization, collective defences and social immunity in the ant Formica selysi. We first describe the diversity and prevalence of fungal pathogens associated with ant nests. The richness of fungal parasites community may increase the risk of multiple infections and select for a diversification of anti-parasitic defences in ants. Collective defences are powerful means to combat parasites, but can also increase the risk of disease transmission. Here, we showed that allo-grooming (mutual cleaning) was directed towards every returning individuals, be they contaminated or not. This collective behaviour removed conidia more efficiently than self-grooming but did not improve the survival of contaminated individuals. This suggests that allo-grooming may rather protect the group than cure contaminated individuals. It may also permit "social vaccination" if a contact with contaminated ants protects groomers frorn a second fungal exposure. Social transfer of immunity is an emerging theme in insect immunology. Here, we showed that ants in contact with an ant from a different genetic lineage had a higher disease resistance. We also found that naïve ants had a higher resistance after a contact with an immunized ant. This suggests that a transfer of resistance is possible and that "social vaccination" may improve the resistance of the group. However, it remains unclear whether repeated exposure to parasites may also increase the resistance of infected individuals themselves. lmmune memory in invertebrates is still debated. We tested whether immune priming against fungal parasite arose in ants and whether it was strain-specific. We found no evidence of immune priming. Naïve and immunized ants had a similar survival when infected. Together with our previous results, this suggests that ants have evolved efficient collective anti-fungal defences but that these defences aim at protecting the group rather than the contaminated individuals. ln colonies of our study population, there is a strong variation in the number of breeders. This is associated with important changes in life-history traits like demography or queen and worker body size. In the second part of the thesis, we investigated how social structures evolved and were maintained. We showed that queens from monogyne and polygyne colonies were able to found new colonies both alone or in association. We also found that there was no difference between monogyne and polygyne colonies in the acceptance of additional queens. These results suggest that a high plasticity has been maintained in this population, which may permit to adapt rapidly to changing environmental conditions. RESUME : Parasites et socialité chez les fourmis Durant cette thèse, nous avons étudié comment la socialité apporte de nouvelles réponses a des problèmes complexes telle que la défense contre les parasites ou l'organisation de la vie en groupe. Nous avons choisi comme modèle la fourmi Formica selysi et ses champignons pathogènes. Nous avons d'abord montré que la diversité et la prévalence de champignons pathogènes associés aux nids de fourmis étaient très élevées. Cela a pu pousser les fourmis à diversifier le champ de leur défenses anti-parasitaires afin d'éviter les infections multiples, La socialité a en particulier permis l'évolution de défenses collectives qui pourraient être plus efficaces que les défenses individuelles. Nous nous sommes donc intéressés de plus près aux défenses collectives et avons étudié quels en étaient les coûts et les bénéfices pour le groupe et pour ses membres. Nous avons trouvé que les fourmis nettoyaient tous les individus entrant dans la colonie, qu'ils soient contaminés ou non. Cela permettait d'ôter plus de spores que le nettoyage individuel et n'augmentait pas la transmission de maladie. Cependant, le nettoyage mutuel n'augmentait pas non plus la survie des individus contaminés. ll se pourrait donc que ce comportement serve plutôt a éviter une dissémination de la maladie qu'à soigner les individus contaminés. Le nettoyage mutuel pourrait aussi permettre aux individus sains d'avoir un premier contact non-létal avec un parasite et d'être vaccinés contre une future exposition. Cette hypothèse a été soutenue par une expérience dans laquelle nous avons montré que le contact avec une fourmi immunisée permettait d'augmenter la résistance d'individus naïfs. Les fourmis avaient aussi une meilleure résistance lorsqu'elles étaient en contact avec une fourmi provenant d'une autre lignée génétique. Cette "vaccination sociale" pourrait permettre d'une part d'augmenter le nombre d'espèce de parasites contre lesquelles le groupe serait protégé et d'autre part de faire l'économie d'autres défenses individuelles telles que la réponse immunitaire. Nous avons testé si les fourmis étaient elles-mêmes "vaccinées", c'est-à-dire, si elles exprimaient une mémoire immunitaire après un premier contact avec un champignon parasite. Nous n'avons trouvé aucune différence de survie entre les individus naïfs et immunisés ce qui suggère les fourmis favorisent d'autres défenses que la mémoire immunitaire contre les champignons entomopathogènes. Cela suggère également que les comportements coopératifs anti-parasitaires pourraient compléter, voire remplacer les défenses individuelles. La socialité telle qu'elle est pratiquée par les fourmis pose un autre problème de poids qui est celui de savoir combien d'individus se reproduisent. En effet, si les ouvrières sont stériles, le nombre de reines assurant la reproduction peut varier considérablement. Dans la population de E sebrsi étudiée, les colonies monogynes (une reine) co-existent avec des colonies polygynes (plusieurs reines) dans le même habitat. Nous nous sommes demandés si ces structures sociales étaient fixes ou si un changement de l'une à l'autre était possible. Pour cela nous avons comparé la fondation de nouvelles colonies par les jeunes reines issues de colonies monogynes et polygynes. Nous avons également observé si l'acceptation de nouvelles reines était possible dans les deux types de colonies. Nous n'avons trouvé aucune différence entre les deux types de colonies. Cela suggère qu'un changement est possible et que l'évolution des structures sociales est un processus dynamique. Cela pourrait être dû à l'habitat particulièrement changeant dans lequel se trouve notre population qui exigerait d'être capable de s'adapter très rapidement a de nouvelles conditions.

Hospital outbreak of methicillin-resistant Staphylococcus aureus caused by a new, possibly hyperepidemic variant from a previously sporadic strain.

Objective: To describe an ongoing outbreak that tripled the annual detection of methicillin-resistant Staphylococcus aureus (MRSA) carriage in a tertiary care hospital. Methods: Active surveillance of MRSA is performed since 20 years in our hospital. Our protocol includes screening of patients transferred from high-incidence health-care institutions or countries, roommates of new MRSA cases, and wards where _2 patients acquired MRSA during the same week. Contact precautions are used for known carriers. PFGE was used for molecular typing until 2004, and was then replaced by Double-Locus Sequence Typing (DLST). Results: A median yearly incidence of 173 new carriers of MRSA was observed from 2002 to 2007. Since September 2008, an increasing number of new cases were observed, mainly as successive clusters limited to distinct wards, reaching a total of 398 until October 2009. The yearly incidence of new cases rose to 275 in 2008 and 613 in 2009. 60% of the cases were due to one strain: DLST 4−4, ST 228, SCCmecI. The incidence of new cases due to the previously predominant strains remained unchanged. The epidemic strain corresponded to a new variant of a clone responsible for a previous outbreak in 2001, and only sporadically isolated (mean of 20 cases/year) since then. A case- control study documented a significant association between acquisition of the epidemic strain and a stay in intensive and intermediary care units, a highest number of internal transfers, but did not identify a point source of transmission. Infection control practices and antibiotic policy had remained unchanged for several years. Compliance with handhygiene as monitored yearly was on the rise. Screening of 313 healthcare workers only found one carrier of the epidemic strain lately in the outbreak. Additional infection control measures were enforced, including screening at ICU admission and discharge with PCR-based rapid test, routine screening for all patients leaving epidemic wards, introduction of PCR-based rapid test for contact tracing, additional working forces for environmental disinfection, and hospital-wide education of healthcare workers. However, the outbreak was still ongoing after 5 months. Conclusions: Factors linked to the dissemination of this new variant in our institution remain undetermined. This unresolved outbreak suggests that this new variant acquired hyperepidemic properties, which calls for further investigations.

The clc element of Pseudomonas sp. strain B13, a genomic island with various catabolic properties.

Pseudomonas sp. strain B13 is a bacterium known to degrade chloroaromatic compounds. The properties to use 3- and 4-chlorocatechol are determined by a self-transferable DNA element, the clc element, which normally resides at two locations in the cell's chromosome. Here we report the complete nucleotide sequence of the clc element, demonstrating the unique catabolic properties while showing its relatedness to genomic islands and integrative and conjugative elements rather than to other known catabolic plasmids. As far as catabolic functions, the clc element harbored, in addition to the genes for chlorocatechol degradation, a complete functional operon for 2-aminophenol degradation and genes for a putative aromatic compound transport protein and for a multicomponent aromatic ring dioxygenase similar to anthranilate hydroxylase. The genes for catabolic functions were inducible under various conditions, suggesting a network of catabolic pathway induction. For about half of the open reading frames (ORFs) on the clc element, no clear functional prediction could be given, although some indications were found for functions that were similar to plasmid conjugation. The region in which these ORFs were situated displayed a high overall conservation of nucleotide sequence and gene order to genomic regions in other recently completed bacterial genomes or to other genomic islands. Most notably, except for two discrete regions, the clc element was almost 100% identical over the whole length to a chromosomal region in Burkholderia xenovorans LB400. This indicates the dynamic evolution of this type of element and the continued transition between elements with a more pathogenic character and those with catabolic properties.

Comprehensive approach for the detection of antifungal compounds using a susceptible strain of Candida albicans and confirmation of in vivo activity with the Galleria mellonella model.

An efficient screening strategy for the identification of potentially interesting low-abundance antifungal natural products in crude extracts that combines both a sensitive bioautography assay and high performance liquid chromatography (HPLC) microfractionation was developed. This method relies on high performance thin layer chromatography (HPTLC) bioautography with a hypersusceptible engineered strain of Candida albicans (DSY2621) for bioactivity detection, followed by the evaluation of wild type strains in standard microdilution antifungal assays. Active extracts were microfractionated by HPLC in 96-well plates, and the fractions were subsequently submitted to the bioassay. This procedure enabled precise localisation of the antifungal compounds directly in the HPLC chromatograms of the crude extracts. HPLC-PDA-mass spectrometry (MS) data obtained in parallel to the HPLC antifungal profiles provided a first chemical screening about the bioactive constituents. Transposition of the HPLC analytical conditions to medium-pressure liquid chromatography (MPLC) allowed the efficient isolation of the active constituents in mg amounts for structure confirmation and more extensive characterisation of their biological activities. The antifungal properties of the isolated natural products were evaluated by their minimum inhibitory concentration (MIC) in a dilution assay against both wild type and engineered strains of C. albicans. The biological activity of the most promising agents was further evaluated in vitro by electron microscopy and in vivo in a Galleria mellonella model of C. albicans infection. The overall procedure represents a rational and comprehensive means of evaluating antifungal activity from various perspectives for the selection of initial hits that can be explored in more in-depth mode-of-action studies. This strategy is illustrated by the identification and bioactivity evaluation of a series of antifungal compounds from the methanolic extract of a Rubiaceae plant, Morinda tomentosa, which was used as a model in these studies.

The region makes the difference: disparities in management of acute myocardial infaction within Switzerland

Purpose: To assess geographical differences within Switzerland regarding management and revascularization procedures for acute myocardial infarction (AMI). Methods: Swiss hospital discharge database for period 2007-2008. The main inclusion criterion was AMI as a primary discharge diagnosis. AMI revascularization procedures were identified and seven Swiss regions (Leman, Mittelland, Northwest, Zurich, Central, Eastern and Ticino) were analyzed. Results: Data from 25,674 AMI discharges were analyzed. Almost half (53.6%) of them were managed in a single hospital, the values ranging from 63.1% (Leman) to 31.4% (Ticino) see table. Relative to the total number of discharges, the highest Intensive Care Unit admission rate was in Leman (69.7%), the lowest (16.4%) in Ticino (Swiss average: 35.8%). Intracoronary revascularization rates were highest in Leman (51.6%) and lowest (30.8%) in Central Switzerland (Swiss average: 41.4%). Bare (non-drug-eluting) stents use was highest in Leman (33.1%) and lowest (7.0%) in Ticino (Swiss average: 15.8%), while drug eluting stent use was highest (32.8%) in Ticino and lowest (13.9%) in Central Switzerland (Swiss average: 24.0%). Coronary artery bypass graft rates were highest (4.6%) in Ticino and lowest (0.4%) in Eastern Switzerland (Swiss average: 2.6%). Mechanical circulatory assistance rates were highest (4.1%) in Zurich and lowest (0.4%) in Ticino (Swiss average: 1.7%). The differences in revascularization procedures remained after adjusting for age, single or multiple hospital management and gender. Conclusion: In Switzerland, significant geographical differences in management and revascularization procedures for AMI were found.

Sex difference in hepatic peroxisome proliferator-activated receptor alpha expression: influence of pituitary and gonadal hormones.

Peroxisome proliferator-activated receptor (PPAR) alpha is a nuclear receptor that is mainly expressed in tissues with a high degree of fatty acid oxidation such as liver, heart, and skeletal muscle. Unsaturated fatty acids, their derivatives, and fibrates activate PPARalpha. Male rats are more responsive to fibrates than female rats. We therefore wanted to investigate if there is a sex difference in PPARalpha expression. Male rats had higher levels of hepatic PPARalpha mRNA and protein than female rats. Fasting increased hepatic PPARalpha mRNA levels to a similar degree in both sexes. Gonadectomy of male rats decreased PPARalpha mRNA expression to similar levels as in intact and gonadectomized female rats. Hypophysectomy increased hepatic PPARalpha mRNA and protein levels. The increase in PPARalpha mRNA after hypophysectomy was more pronounced in females than in males. GH treatment decreased PPARalpha mRNA and protein levels, but the sex-differentiated secretory pattern of GH does not determine the sex-differentiated expression of PPARalpha. The expression of PPARalpha mRNA in heart or soleus muscle was not influenced by gender, gonadectomy, hypophysectomy, or GH treatment. In summary, pituitary-dependent hormones specifically regulate hepatic PPARalpha expression. Sex hormones regulate the sex difference in hepatic PPARalpha levels, but not via the sexually dimorphic GH secretory pattern.