Papel del lípido A de Klebsiella pneumoniae en el control de la respuesta inmune.

Klebsiella pneumoniae és un bacil Gram negatiu que causa infeccions del tracte urinari així com pneumònies adquirides en la comunitat i nosocomials. La membrana externa de les bactèries Gram negatives està formada pel lipopolisacàrid (LPS). El LPS és una molècula formada per un polisacàrid, amb càrrega negativa, ancorat a una regió hidrofòbica, el lípid A, que posseeix característiques d’endotoxina i és, per tant, immunoestimuladora. Els sistemes de transducció de senyals de dos components en bactèries detecten canvies en l’ambient i regulen l’expressió de gens que modifiquen l’estructura del lípid A per tal de mantenir l’homeòstasi de la cèl•lula bacteriana, augmentar la resistència en front als pèptids catiònics o modular la resposta immune de l’hoste. En resultats previs del nostre laboratori va posar-se de manifest que la xaperona Hfq regula les modificacions del lípid A a Yersinia enterocolitica O8. En aquesta tesi doctoral posem de manifest que aquest fet no sols es dóna a Y. enterocolitica si no que Hfq també participa en la regulació de les modificacions dels lípid A tant a Salmonella Typhimurium com a K. pneumoniae. En la segona part d’aquesta tesi doctoral hem caracteritzat l’estructura de lípid A que K. pneumoniae expressa in vivo, durant una infecció, sense la necessitat de realitzar un pas previ d’aïllament bacterià en medis de cultiu tradicionals. Durant una infecció pulmonar, K. pneumoniae expressa un lípid A que es caracteritza per la presència d’una cadena de 2-hidroximiristat com acilació secundària. Les nostres observacions indiquen que el lípid A que K. pneumoniae expressa en el pulmó és menys inflamatori que el lípid A que la bactèria expressa quan creix en medis de cultiu tradicionals. A més a més, el lípid A que K. pneumoniae expressa in vivo juga un importantíssim paper conferint a la bactèria resistència en front als pèptids antimicrobiansKlebsiella pneumoniae es un bacilo Gram negativo que causa infecciones del tracto urinario así como neumonías adquiridas en la comunidad y nosocomiales. La membrana externa de las bacterias Gram negativas se compone del lipopolisacárido (LPS). El LPS es una molécula formada por un polisacárido, cargado negativamente, anclado a un dominio hidrofóbico, el lípido A, que posee propiedades de endotoxina y, por tanto, es inmunoestimulador. Los sistemas de transducción de señales de dos componentes en bacterias detectan cambios en el ambiente y regulan la expresión de genes que remodelan la estructura del lípido A para mantener la homeóstasis de la célula bacteriana, aumentar su resistencia a péptidos catiónicos o modular la respuesta inmune del huésped. En resultados previos de nuestro laboratorio se puso de manifiesto que la chaperona Hfq participa en la regulación de las modificaciones del lípido A en Yersinia enterocolitica. En esta tesis doctoral se ha puesto de manifiesto que este hecho no sólo ocurre en Y. enterocolitica si no que también Hfq participa en la regulación de las modificaciones del lípido A tanto en Salmonella Typhimurium como en K. pneumoniae. En la segunda parte de esta tesis doctoral se ha caracterizado la estructura de lípido A que expresa K. pneumoniae in vivo durante una infección sin la necesidad de aislar la bacteria previamente en medios de cultivo tradicionales. Durante una infección pulmonar, K. pneumoniae expresa una estructura de lípido A caracterizada por la presencia de una cadena de 2-hidroximiristato como acilación secundaria. Nuestras observaciones indican que el lípido A expresado en el pulmón es menos inflamatorio que el expresado cuando la bacteria crece en medios de cultivo tradicionales. Además, el lípido A expresado por K. pneumoniae in vivo juega un importante papel en la resistencia de la bacteria frente a péptidos antimicrobianos.K. pneumoniae is a Gram negative bacterium which causes urinary tract infections as well as community-acquired and nosocomial pneumonia. Lipopolysaccharide (LPS) is the major component of the outer membrane of Gram negative bacteria. LPS is a negatively charged polysaccharide anchored to a hydrophobic region, the endotoxic lipid A. In bacteria, two component transduction systems detect environmental changes and trigger the expression of genes responsible for lipid A modifications. Lipid A remodelling helps maintaining cellular integrity, confers resistance to many antimicrobial peptides as well as modulates host’s immune response. Previous results from our laboratory showed that Hfq regulates lipid A modifications in Yersinia enterocolitica O8. In this doctoral thesis we demonstrate that Hfq also regulates lipid A modifications in other Enterobacteriaceae such as Salmonella Typhimurium and K. pneumoniae. In a second study, we have characterised for the first time the lipid A structure expressed by K. pneumoniae in vivo during a lung infection without the need of a previous isolation step on traditional culture media. During the course of a lung infection, K. pneumoniae expresses a lipid A characterised by the presence of a 2-hydroxymiristate as a secondary acylation. Our results show that the lipid A expressed by K. pneumoniae in vivo in the lung is less inflammatory than the lipid A expressed when cultured on traditional culture media. Furthermore, this lipid A found in the lung plays a major role conferring resistance to antimicrobial peptides

Estimating with kernel smoothers the mean of functional data in a finite population setting. A note on variance estimation in presence of partially observed trajectories

In the near future, millions of load curves measuring the electricity consumption of French households in small time grids (probably half hours) will be available. All these collected load curves represent a huge amount of information which could be exploited using survey sampling techniques. In particular, the total consumption of a specific cus- tomer group (for example all the customers of an electricity supplier) could be estimated using unequal probability random sampling methods. Unfortunately, data collection may undergo technical problems resulting in missing values. In this paper we study a new estimation method for the mean curve in the presence of missing values which consists in extending kernel estimation techniques developed for longitudinal data analysis to sampled curves. Three nonparametric estimators that take account of the missing pieces of trajectories are suggested. We also study pointwise variance estimators which are based on linearization techniques. The particular but very important case of stratified sampling is then specifically studied. Finally, we discuss some more practical aspects such as choosing the bandwidth values for the kernel and estimating the probabilities of observation of the trajectories.Comment: Version revised for Statistics and Probability Letter

A QM/MM study on the origin of retro-aldolase activity of a catalytic antibody

The retro-aldolase mechanism of methodol catalysed by the catalytic antibody 33F12 is described based on the exploration of the free energy landscape obtained with QM/MM methods. The amino acids involved in the reaction have been identified, as well as their specific role played in the active site and in the flexibility of the loops. Finally, the comparison with a de novo enzyme RA95.5-8F provides a deeper understanding of catalytic differences between such different protein scaffolds

Understanding the Directed Evolution of De Novo Retro-Aldolases from QM/MM Studies

In an era in which climatic change puts the planet at risk, the study and development of alternative green chemistry which can help and improve our life can play an essential role. In this context, the use of artificial enzymes that are capable of substitute traditional industrial processes by environmental friendly routes is a challenge. Unfortunately, the complete understanding of the catalytic activity and selectivity of enzymes remains elusive, thus hampering creation and development of enzymatic proteins. In this paper, the molecular mechanism of the non-natural multistep retro-aldolase reaction catalyzed by a de novo biocatalyst, the RA95.5-5, has been investigated by means of multiscale QM/MM methods. The design of a retro-aldolase presents the difficulty to create an enzyme being able to stabilize several transition states, maintaining low-energy barriers along the overall reaction. The obtained QM/MM free-energy landscape has allowed defining the rate-determining step corresponding to the carbon–carbon bond scission of the substrate, which is in accordance with the experimental data. A deep analysis of the electrostatic interactions between the substrate and the different amino acid residues of the protein, as well as the estimation of the electrostatic potential generated on key atoms of the substrate, has been carried out for the key steps of the reaction. The results, compared with previous computational studies on the most efficient de novo retro-aldolase, the RA95.5-8F, explains the different activities achieved during the directed evolution process and provides insights for future developments of more efficient enzymes

Measurement and prediction of quantum coherence effects in biological processes

This themed issue presents a collection of articles on the measurement and prediction of quantum coherence effects in biological processes.</p

Determination of the microbiological quality and degree of transfer of Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium and Enteritidis in unpasteurized juices from supermarkets

PòsterJun

Seeing the last part of a hitting movement is enough to adapt to a temporal delay.

Being able to see the object that you are aiming for is evidently useful for guiding the hand to a moving object. We examined to what extent seeing the moving hand also influences performance. Subjects tried to intercept moving targets while either instantaneous or delayed feedback about the moving hand was provided at certain times. After each attempt, subjects had to indicate whether they thought they had hit the target, had passed ahead of it, or had passed behind it. Providing visual feedback early in the movement enabled subjects to use visual information about the moving hand to correct their movements. Providing visual feedback when the moving hand passed the target helped them judge how they had performed. Performance was almost as good when visual feedback about the moving hand was provided only when the hand was passing the target as when it was provided throughout the movement. We conclude that seeing the temporal relationship between the hand and the target as the hand crosses the target's path is instrumental for adapting to a temporal delay

Dealing with delays does not transfer across sensorimotor tasks.

It is known that people can learn to deal with delays between their actions and the consequences of such actions. We wondered whether they do so by adjusting their anticipations about the sensory consequences of their actions or whether they simply learn to move in certain ways when performing specific tasks. To find out, we examined details of how people learn to intercept a moving target with a cursor that follows the hand with a delay and examined the transfer of learning between this task and various other tasks that require temporal precision. Subjects readily learned to intercept the moving target with the delayed cursor. The compensation for the delay generalized across modifications of the task, so subjects did not simply learn to move in a certain way in specific circumstances. The compensation did not generalize to completely different timing tasks, so subjects did not generally expect the consequences of their motor commands to be delayed. We conclude that people specifically learn to control the delayed visual consequences of their actions to perform certain tasks

Modelado de una cámara de onda milimétrica

En aquest projecte s'ha dissenyat i simulat una càmera d'ones mil·limètriques mitjançant "Spinning Reflectors" amb l'objectiu d'estudiar la qualitat de les imatges obtingudes, així com determinar la configuració de reflectors acceptable per a la nostra càmera. Per al seu disseny, s'ha realitzat el model d'una antena Cassegrain amb botzina alimentadora a FEKO i s'han simulat els diagrames de radiació corresponents als diferents angles de rotació d'ambdós reflectors al voltant de l'eix en el qual estan situats. Finalment, a partir del principi de funcionament de la nostra càmera, s'ha exposat la seva implementació física i s'ha representat en Matlab la seva PSF (Point Spread Function) i la imatge d'un objecte arbitrari d'acord amb les tècniques de formació d'imatges.En este proyecto se ha diseñado y simulado una cámara de ondas milimétricas mediante "Spinning Reflectors" con el objetivo de estudiar la calidad de las imágenes obtenidas, así como determinar la configuración de reflectores aceptable para nuestra cámara. Para su diseño, se ha realizado el modelo de una antena Cassegrain con bocina alimentadora en FEKO y se han simulado los diagramas de radiación correspondientes a los diferentes ángulos de rotación de ambos reflectores alrededor del eje en el que están situados. Finalmente, a partir del principio de funcionamiento de nuestra cámara, se ha expuesto su implementación física y se ha representado en Matlab su PSF (Point Spread Function) y la imagen de un objeto arbitrario de acuerdo con las técnicas de formación de imágenes.In this project we have designed and simulated a millimeter wave camera using "Spinning Reflectors" in order to study the quality of the images and to determine the acceptable configuration of reflectors to our camera. For his design, we have performed a model of a Cassegrain antenna with horn feed in FEKO, and we have simulated the radiation patterns corresponding to the different angles of rotation of the two reflectors around the axis on which they are located. Finally, agree with the operating principle of our camera, we have exposed her physical implementation and we have represented in Matlab her PSF (Point Spread Function) and the image of an arbitrary object in accordance with the imaging techniques