628 resultados para LAU


Relevância:

10.00% 10.00%

Publicador:

Resumo:

The Drive for Muscularity Scale (DMS) is a widely used measure in studies of men’s body image, but few studies have examined its psychometric properties outside English-speaking samples. Here, we assessed the factor structure of a Malay translation of the DMS. A community sample of 159 Malay men from Kuala Lumpur, Malaysia, completed the DMS, along with measures of self-esteem, body appreciation, and muscle discrepancy. Exploratory factor analysis led to the extraction of two factors, differentiating attitudes from behaviours, which mirrors the parent scale. Both factors also loaded on to a higher-order drive for muscularity factor. The subscales of the Malay DMS had adequate internal consistencies and good convergent validity, insofar as significant relationships were reported with self-esteem, body appreciation,muscle discrepancy, and body mass index. These results indicate that the Malay DMS has acceptable psychometric properties and can be used to assess body image concerns in Malay men.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Este trabajo tiene como objetivo estudiar los diferentes estados mentales de los personajes de las novelas de Tu rostro mañana, de Javier Marías. Estudiando las reflexiones del narrador sobre J. Deza, Peter Wheeler o Francisco Rico observamos que su decadencia mental se muestra a través de una suerte de ―presciencia‖ o lucidez momentánea que puede servir para mostrar el silencio como única tendencia de todo discurso. Desde el momento en que toda historia de ficción se cimenta sobre un discurso –no importa su cauce de presentación, ni su fuente– este es falsificado por el tiempo, la gente y cualquier otra herramienta que pueda ser utilizada para contar nada. Las conclusiones de este trabajo muestran la quimera que implica tratar de mantener una contención absoluta sobre lo acaecido, pues dicho vacío de narrativas será ocupado por una suplantación que suele ser el reverso más infame de sus actores. Es por ello que el narrador J. Deza sigue conminado a explicar sus historias, incluso allí donde uno diría que ya no puede haber ni palabras suficientes para traducir un hecho en ficción.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

The McMurdo Dry Valleys of Antarctica are an extreme polar desert. Mineral soils support subsurface microbial communities and translucent rocks support development of hypolithic communities on ventral surfaces in soil contact. Despite significant research attention, relatively little is known about taxonomic and functional diversity or their inter-relationships. Here we report a combined diversity and functional interrogation for soil and hypoliths of the Miers Valley in the McMurdo Dry Valleys of Antarctica. The study employed 16S rRNA fingerprinting and high throughput sequencing combined with the GeoChip functional microarray. The soil community was revealed as a highly diverse reservoir of bacterial diversity dominated by actinobacteria. Hypolithic communities were less diverse and dominated by cyanobacteria. Major differences in putative functionality were that soil communities displayed greater diversity in stress tolerance and recalcitrant substrate utilization pathways, whilst hypolithic communities supported greater diversity of nutrient limitation adaptation pathways. A relatively high level of functional redundancy in both soil and hypoliths may indicate adaptation of these communities to fluctuating environmental conditions.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Los principales recursos pesqueros pelágicos de interés económico en el Perú son anchoveta (Engraulis ringens), jurel (Trachurus murphyi) y caballa (Scomber japonicus) [3]. Para su evaluación, se lleva a cabo cruceros de evaluación acústica en los que se integra información de ecoabundancia y proporción de tallas por especie para obtener valores de biomasa y abundancia. Sin embargo, para especies no objetivo (como jurel), dichos valores resultan poco confiables por la lejanía entre los puntos de muestreo biométrico y acústico. Para resolver este inconveniente, el presente trabajo propuso utilizar modelos empíricos (de tipo GAM y GLM) integrando variables ambientales y de seguimiento de desembarques con la finalidad de generar índices relativos y absolutos para anchoveta y jurel en el período de 1996-2013 dentro del área de las 200 mn frente a la costa peruana. Los resultados obtenidos realzaron la importancia de los lances de comprobación para la obtención de estimaciones robustas de biomasa. Así mismo, se observó que, para anchoveta, los modelos empíricos sí produjeron un buen índice relativo y absoluto, mejorando la utilización de la ecoabundancia por sí sola. Para jurel, sin embargo, el modelo final calibrado resultó en la obtención de un mejor índice relativo. Se recomienda además, la obtención de información de tallas y pesos medios de desembarques para jurel con la finalidad de mejorar las estimaciones de biomasa y abundancia.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

L’adversité tôt dans la vie est associée au développement de symptômes anxieux pouvant perdurer jusqu’à l’âge adulte (Casey et el, 2010, Pine 2003). Des études chez l’adulte suggèrent que ces liens pourraient être associés à des altérations du « circuit de la peur » qui inclut l’amygdale, l’hippocampe antérieur, l’insula et le cortex préfrontal (Marek, 2013, Etkin & Wager, 2007). Ceci a cependant peu été étudié chez les jeunes. L’objectif principal de cette thèse était de définir les corrélats comportementaux, physiologiques, biologiques et neuronaux du traitement de la peur chez les jeunes en bonne santé, en lien ou non avec un historique d’adversité -- sous la forme de pratiques parentales coercitives -- et d’anxiété. D’abord, puisque nous nous intéressions aux pratiques parentales coercitives chroniques, nous avons examiné leur évolution et facteurs de risque, en nous concentrant sur la période de 17 à 72 mois. Un total de 2045 dyades mère-enfant ont été incluses dans une analyse de courbe de croissance latente. Nous avons démontré que la coercition maternelle suit une évolution non linéaire durant cette période et atteint un sommet à 42 mois. Les facteurs de risque relatifs à l’enfant et à la mère, mesurés à 17 mois, permettent de prédire les niveaux de coercition à 42 mois. Finalement, les prédicteurs relatifs à l’enfant et l’efficacité maternelle prédisent l’évolution des pratiques parentales coercitives entre 17 et 72 mois. Ensuite, afin de définir une méthodologie solide pour étudier le traitement de la peur chez des jeunes, nous avons adapté une tâche développée par Lau et ses collaborateurs (2008), employant des visages féminins comme stimuli. Le sexe des participants et des visages employés comme stimuli pouvant potentiellement moduler le traitement de la peur (Kret & de Gelder, 2012; McClure, 2000), nous avons étudié leurs influences respectives sur les réponses électrodermales et subjectives de peur durant le conditionnement et l’extinction de la peur chez 117 jeunes. Nous avons démontré que les stimuli féminins suscitent des réponses davantage comparables entre les garçons et les filles que les stimuli masculins. De plus, nous avons observé un effet du « même sexe », caractérisé par un conditionnement différentiel uniquement face aux stimuli du même sexe que le participant. Finalement, nous avons exploré les différences individuelles et conjointes associées aux différents niveaux de pratiques parentales coercitives et d’anxiété en termes de réponses de peur et d’activité cérébrale, durant le conditionnement et l’extinction de la peur chez 84 jeunes. Nous avons démontré que la coercition est spécifiquement associée au fonctionnement du lobe temporal médian et aux interactions entre l’amygdale et l’insula, durant le conditionnement. Durant l’extinction, les niveaux d’anxiété étaient associés à des différences spécifiques d’activation du gyrus cingulaire antérieur (GCA) dorsal. Enfin, les pratiques parentales coercitives et l’anxiété interagissent et viennent moduler la connectivité fonctionnelle amygdale - GCA rostral, l’activation d’une sous-région du GCA dorsal et les réponses subjectives de peur. Ces résultats ajoutent une pièce au casse-tête des neurosciences développementales et fournissent des pistes intéressantes pour le développement d’interventions futures.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Hallux rígidus (HR) affects the first metatarsophalangeal joint (MTPJ) between 35% and 60% of the population over 65 years and there are multiple ways of treatment. Depending on the radiological stage where you find the deformity determines the procedure to be performed; in the early stages cheilectomy techniques and corrective osteotomy is performed while the more advanced ratings, the surgeon chooses destructive techniques considered as arthrodesis and arthroplasty. This final of degree project aims to focus on 1 MTPJ destructive techniques to clarify which of the procedures generates better results by a number of parameters; outcomes of the American Orthopaedic Foot scale and Ankle Society Hallux metatarsophalangeal Interphalangeal-scale (AOFAS), range of motion (ROM) of the 1ºAMTF, radiological classification. As for the implant arthroplasty technique, this article offers information on material and design that generates better relating to patient characteristics such as age, inflammatory joint diseases, viability and durability of the implant results. The conclusion from this review is that the values obtained in the arthrodesis according AOFAS decrease due to loss of mobility, but both techniques have similar values of effectiveness and concludes with the decision that the technique used is determined taking into account various factors and patient characteristics. Keywords: Hallux rígidus; (Hallux Rígidus) and surgery treatment; Hallux Rígidus arthrodesis; Hallux Rígidus arthroplasty; Hallux Rígidus (arthroplasty and arthrodesis).

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Le développement durable et la participation citoyenne sont deux termes très en vogue actuellement. Ils sont utilisés de bien des manières et même parfois de façon détournée. Néanmoins, il est de plus en plus fréquent de voir la participation citoyenne et le développement durable être associés. Cela provient du fait que pour les tenants de cette fusion, pour qu’il y ait développement durable il doit y avoir une forme de participation citoyenne. Ce lien théorique entre les deux concepts est vrai pour ses adhérents puisque ce doit être les citoyens impliqués dans le développement qui doivent guider en partie l’élaboration de la politique de développement durable. Les politiques d’urbanismes sont parmi les politiques les plus importantes pour créer un développement durable puisqu’elle combine à la fois le transport et l’aménagement du territoire selon une perspective économique, sociale et environnementale. De plus, ce sont les politiques d’urbanismes qui gèrent les milieux naturels. Pour Gauthier et Gagnon (2013) afin qu’une politique d’urbanisme durable émerge il doit y avoir une forme spécifique de participation citoyenne soit une participation citoyenne significative. Le Schéma d’aménagement et de développement de la Ville de Sherbrooke est en fait une politique d’urbanisme tendant vers le développement durable qui a inclus une démarche de participation citoyenne. Une grille d’analyse basée sur les travaux de M. Gauthier et de Mme Gagnon a permis de savoir si l'outil de participation citoyenne utilisée par la Ville de Sherbrooke dans le cadre de l’élaboration de son schéma d’aménagement et de développement adopté en 2013 a permis une participation citoyenne significative selon les huit critères établis par Mario Gauthier et Lynda Gagnon (2013)? Deux citoyens, une élue de la Ville, une fonctionnaire responsable du Schéma et trois représentants d’associations ont donné leurs perceptions dans des entrevues semi-dirigées afin de qualifier la participation citoyenne lors de l’élaboration du Schéma d’aménagement de la Ville. Les perceptions des participants n’ont pas permis de qualifier le processus participatif de la Ville. Il est donc impossible pour le chercheur de confirmer si la démarche de participation citoyenne est significative ou non. La Loi sur l’urbanisme et l’aménagement du Québec (LAU) a influencé de façon non significative la participation citoyenne lors de l’élaboration du Schéma. Il est clair que celle-ci a une influence en instituant de façon obligatoire la participation citoyenne lors de l’élaboration de Schéma d’aménagement et de développement des MRC, mais malgré les critères obligatoires de bases ce n’est pas la LAU qui a influencé de manière significative la participation citoyenne dans la forme analysée.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

El Cermi recibió en 2011 el mandato de realizar el seguimiento de la aplicación de la Convención Internacional sobre los Derechos de las Personas con Discapacidad en España. El documento que se reseña aquí corresponde al informe de 2012 y consta de cuatro partes. En la primera, se repasa la Convención artículo por artículo, señalándose las vulneraciones, buenas prácticas y resoluciones judiciales relativas a cada uno de ellos, así como propuestas de mejora. El segundo capítulo analiza estadísticamente las quejas recibidas por el Cermi y hace balance de la actuación de esta ONG en la defensa de los derechos de las personas con discapacidad. El tercer apartado del documento examina los informes presentados en torno a esta cuestión por la Oficina Permanente Especializada del Consejo Nacional de la Discapacidad, y los Defensores del Pueblo estatal y autonómicos. Finalmente, se recoge el informe de la Clínica Jurídica llevada a cabo por el Instituto de Derechos Humanos Bartolomé de las Casas en torno al trato dispensado a un joven de nacionalidad marroquí con discapacidad sobrevenida.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

This thesis addresses the Batch Reinforcement Learning methods in Robotics. This sub-class of Reinforcement Learning has shown promising results and has been the focus of recent research. Three contributions are proposed that aim to extend the state-of-art methods allowing for a faster and more stable learning process, such as required for learning in Robotics. The Q-learning update-rule is widely applied, since it allows to learn without the presence of a model of the environment. However, this update-rule is transition-based and does not take advantage of the underlying episodic structure of collected batch of interactions. The Q-Batch update-rule is proposed in this thesis, to process experiencies along the trajectories collected in the interaction phase. This allows a faster propagation of obtained rewards and penalties, resulting in faster and more robust learning. Non-parametric function approximations are explored, such as Gaussian Processes. This type of approximators allows to encode prior knowledge about the latent function, in the form of kernels, providing a higher level of exibility and accuracy. The application of Gaussian Processes in Batch Reinforcement Learning presented a higher performance in learning tasks than other function approximations used in the literature. Lastly, in order to extract more information from the experiences collected by the agent, model-learning techniques are incorporated to learn the system dynamics. In this way, it is possible to augment the set of collected experiences with experiences generated through planning using the learned models. Experiments were carried out mainly in simulation, with some tests carried out in a physical robotic platform. The obtained results show that the proposed approaches are able to outperform the classical Fitted Q Iteration.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Lundi matin. Le réveil claironne. Il faut rassembler toutes ses énergies pour s'attaquer à une autre semaine et ses quarante heures de boulot, et cette perspective sourira aux uns et affligera les autres, selon l'humeur du moment. Après quelques années de participation à la population active, ces habitudes et opinions s'ancrent profondément en nous pour relever finalement de l'automatisme. On pourrait ainsi énumérer une longue liste d'actes et d'attitudes qui sont passés à l'inconscient tellement la routine s'est installée. Cependant, une personne ne naît pas avec des habitudes de travail et des opinions à ce sujet. Elle les acquière au fil des années. En effet, il fût un jour où elle n'avait pas à traiter avec des éléments tels les syndicats, les patrons, le chômage, etc. Elle fréquentait encore l'école, lieu d'apprentissage où, malgré tout, elle possédait sûrement quelques opinions au sujet des éléments précités. Questionnons l'élève pour connaître ses attitudes vis-à-vis la prochaine étape de sa vie, étape qui durera une quarantaine d'années et qui débouchera sur une retraite, active ou non. Connaître les attitudes des élèves - plus précisément celles des finissants du secteur professionnel des paliers secondaire et collégial – à propos du monde du travail, voilà, en gros, la ligne directrice de ce travail de recherche. Pour ce faire, nous nous sommes tournés vers environ 2 000 élèves de l'Estrie qui, en 1983, étaient inscrits aux paliers éducationnels qui forment soit des ouvriers soit des techniciens. L'analyse détaillée des résultats du questionnaire nous offre un regard sur maints éléments sociologiques fort intéressants. Ainsi pourrons-nous découvrir certains éléments de l'idéologie qui sera celle d'une importante composante du bassin démographique québécois de l'avenir. Au Québec, entre autres, plusieurs intervenants politiques, économiques et sociaux ne prennent pas le temps de réfléchir aux répercussions des conditions sociales actuelles sur les jeunes qui s'apprêtent à tenter de trouver une niche au sein du marché du travail Lors de votre première journée à la maternelle, on a mis en oeuvre tous les moyens inimaginables pour que votre intégration se fasse en douceur. En fait-on autant pour la personne qui s'apprête à relever un défi long de quarante ans? Que lui arrivera-t-elle si on la traumatise dès le début? Comment auriez-vous réagi si on vous avait refusé l'accès à la maternelle? Mettez-vous dans la peau du jeune chômeur chronique et les réponses viendront aisément. Enfin, tout ceci pour dire que le finissant possède assurément une idée de ce qui l'attend dans l’«au-delà» de son école polyvalente. Mais cette idée concorde-t-elle avec celle du finissant du collégial? En quels points ces idées se rejoignent-elles? En quels autres divergent-elles? Y a-t-il un fil conducteur? Les questions sont posées. Tentons d'y répondre au moyen de la présente recherche.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Robotics is an emergent branch of engineering that involves the conception, manufacture, and control of robots. It is a multidisciplinary field that combines electronics, design, computer science, artificial intelligence, mechanics and nanotechnology. Its evolution results in machines that are able to perform tasks with some level of complexity. Multi-agent systems is a researching topic within robotics, thus they allow the solving of higher complexity problems, through the execution of simple routines. Robotic soccer allows the study and development of robotics and multiagent systems, as the agents have to work together as a team, having in consideration most problems found in our quotidian, as for example adaptation to a highly dynamic environment as it is the one of a soccer game. CAMBADA is the robotic soccer team belonging to the group of research IRIS from IEETA, composed by teachers, researchers and students of the University of Aveiro, which annually has as main objective the participation in the RoboCup, in the Middle Size League. The purpose of this work is to improve the coordination in set pieces situations. This thesis introduces a new behavior and the adaptation of the already existing ones in the offensive situation, as well as the proposal of a new positioning method in defensive situations. The developed work was incorporated within the competition software of the robots. Which allows the presentation, in this dissertation, of the experimental results obtained, through simulation software as well as through the physical robots on the laboratory.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Membrane proteins, which reside in the membranes of cells, play a critical role in many important biological processes including cellular signaling, immune response, and material and energy transduction. Because of their key role in maintaining the environment within cells and facilitating intercellular interactions, understanding the function of these proteins is of tremendous medical and biochemical significance. Indeed, the malfunction of membrane proteins has been linked to numerous diseases including diabetes, cirrhosis of the liver, cystic fibrosis, cancer, Alzheimer's disease, hypertension, epilepsy, cataracts, tubulopathy, leukodystrophy, Leigh syndrome, anemia, sensorineural deafness, and hypertrophic cardiomyopathy.1-3 However, the structure of many of these proteins and the changes in their structure that lead to disease-related malfunctions are not well understood. Additionally, at least 60% of the pharmaceuticals currently available are thought to target membrane proteins, despite the fact that their exact mode of operation is not known.4-6 Developing a detailed understanding of the function of a protein is achieved by coupling biochemical experiments with knowledge of the structure of the protein. Currently the most common method for obtaining three-dimensional structure information is X-ray crystallography. However, no a priori methods are currently available to predict crystallization conditions for a given protein.7-14 This limitation is currently overcome by screening a large number of possible combinations of precipitants, buffer, salt, and pH conditions to identify conditions that are conducive to crystal nucleation and growth.7,9,11,15-24 Unfortunately, these screening efforts are often limited by difficulties associated with quantity and purity of available protein samples. While the two most significant bottlenecks for protein structure determination in general are the (i) obtaining sufficient quantities of high quality protein samples and (ii) growing high quality protein crystals that are suitable for X-ray structure determination,7,20,21,23,25-47 membrane proteins present additional challenges. For crystallization it is necessary to extract the membrane proteins from the cellular membrane. However, this process often leads to denaturation. In fact, membrane proteins have proven to be so difficult to crystallize that of the more than 66,000 structures deposited in the Protein Data Bank,48 less than 1% are for membrane proteins, with even fewer present at high resolution (< 2Å)4,6,49 and only a handful are human membrane proteins.49 A variety of strategies including detergent solubilization50-53 and the use of artificial membrane-like environments have been developed to circumvent this challenge.43,53-55 In recent years, the use of a lipidic mesophase as a medium for crystallizing membrane proteins has been demonstrated to increase success for a wide range of membrane proteins, including human receptor proteins.54,56-62 This in meso method for membrane protein crystallization, however, is still by no means routine due to challenges related to sample preparation at sub-microliter volumes and to crystal harvesting and X-ray data collection. This dissertation presents various aspects of the development of a microfluidic platform to enable high throughput in meso membrane protein crystallization at a level beyond the capabilities of current technologies. Microfluidic platforms for protein crystallization and other lab-on-a-chip applications have been well demonstrated.9,63-66 These integrated chips provide fine control over transport phenomena and the ability to perform high throughput analyses via highly integrated fluid networks. However, the development of microfluidic platforms for in meso protein crystallization required the development of strategies to cope with extremely viscous and non-Newtonian fluids. A theoretical treatment of highly viscous fluids in microfluidic devices is presented in Chapter 3, followed by the application of these strategies for the development of a microfluidic mixer capable of preparing a mesophase sample for in meso crystallization at a scale of less than 20 nL in Chapter 4. This approach was validated with the successful on chip in meso crystallization of the membrane protein bacteriorhodopsin. In summary, this is the first report of a microfluidic platform capable of performing in meso crystallization on-chip, representing a 1000x reduction in the scale at which mesophase trials can be prepared. Once protein crystals have formed, they are typically harvested from the droplet they were grown in and mounted for crystallographic analysis. Despite the high throughput automation present in nearly all other aspects of protein structure determination, the harvesting and mounting of crystals is still largely a manual process. Furthermore, during mounting the fragile protein crystals can potentially be damaged, both from physical and environmental shock. To circumvent these challenges an X-ray transparent microfluidic device architecture was developed to couple the benefits of scale, integration, and precise fluid control with the ability to perform in situ X-ray analysis (Chapter 5). This approach was validated successfully by crystallization and subsequent on-chip analysis of the soluble proteins lysozyme, thaumatin, and ribonuclease A and will be extended to microfluidic platforms for in meso membrane protein crystallization. The ability to perform in situ X-ray analysis was shown to provide extremely high quality diffraction data, in part as a result of not being affected by damage due to physical handling of the crystals. As part of the work described in this thesis, a variety of data collection strategies for in situ data analysis were also tested, including merging of small slices of data from a large number of crystals grown on a single chip, to allow for diffraction analysis at biologically relevant temperatures. While such strategies have been applied previously,57,59,61,67 they are potentially challenging when applied via traditional methods due to the need to grow and then mount a large number of crystals with minimal crystal-to-crystal variability. The integrated nature of microfluidic platforms easily enables the generation of a large number of reproducible crystallization trials. This, coupled with in situ analysis capabilities has the potential of being able to acquire high resolution structural data of proteins at biologically relevant conditions for which only small crystals, or crystals which are adversely affected by standard cryocooling techniques, could be obtained (Chapters 5 and 6). While the main focus of protein crystallography is to obtain three-dimensional protein structures, the results of typical experiments provide only a static picture of the protein. The use of polychromatic or Laue X-ray diffraction methods enables the collection of time resolved structural information. These experiments are very sensitive to crystal quality, however, and often suffer from severe radiation damage due to the intense polychromatic X-ray beams. Here, as before, the ability to perform in situ X-ray analysis on many small protein crystals within a microfluidic crystallization platform has the potential to overcome these challenges. An automated method for collecting a "single-shot" of data from a large number of crystals was developed in collaboration with the BioCARS team at the Advanced Photon Source at Argonne National Laboratory (Chapter 6). The work described in this thesis shows that, even more so than for traditional structure determination efforts, the ability to grow and analyze a large number of high quality crystals is critical to enable time resolved structural studies of novel proteins. In addition to enabling X-ray crystallography experiments, the development of X-ray transparent microfluidic platforms also has tremendous potential to answer other scientific questions, such as unraveling the mechanism of in meso crystallization. For instance, the lipidic mesophases utilized during in meso membrane protein crystallization can be characterized by small angle X-ray diffraction analysis. Coupling in situ analysis with microfluidic platforms capable of preparing these difficult mesophase samples at very small volumes has tremendous potential to enable the high throughput analysis of these systems on a scale that is not reasonably achievable using conventional sample preparation strategies (Chapter 7). In collaboration with the LS-CAT team at the Advanced Photon Source, an experimental station for small angle X-ray analysis coupled with the high quality visualization capabilities needed to target specific microfluidic samples on a highly integrated chip is under development. Characterizing the phase behavior of these mesophase systems and the effects of various additives present in crystallization trials is key for developing an understanding of how in meso crystallization occurs. A long term goal of these studies is to enable the rational design of in meso crystallization experiments so as to avoid or limit the need for high throughput screening efforts. In summary, this thesis describes the development of microfluidic platforms for protein crystallization with in situ analysis capabilities. Coupling the ability to perform in situ analysis with the small scale, fine control, and the high throughput nature of microfluidic platforms has tremendous potential to enable a new generation of crystallographic studies and facilitate the structure determination of important biological targets. The development of platforms for in meso membrane protein crystallization is particularly significant because they enable the preparation of highly viscous mixtures at a previously unachievable scale. Work in these areas is ongoing and has tremendous potential to improve not only current the methods of protein crystallization and crystallography, but also to enhance our knowledge of the structure and function of proteins which could have a significant scientific and medical impact on society as a whole. The microfluidic technology described in this thesis has the potential to significantly advance our understanding of the structure and function of membrane proteins, thereby aiding the elucidation of human biology, the development of pharmaceuticals with fewer side effects for a wide range of diseases. References (1) Quick, M.; Javitch, J. A. P Natl Acad Sci USA 2007, 104, 3603. (2) Trubetskoy, V. S.; Burke, T. J. Am Lab 2005, 37, 19. (3) Pecina, P.; Houstkova, H.; Hansikova, H.; Zeman, J.; Houstek, J. Physiol Res 2004, 53, S213. (4) Arinaminpathy, Y.; Khurana, E.; Engelman, D. M.; Gerstein, M. B. Drug Discovery Today 2009, 14, 1130. (5) Overington, J. P.; Al-Lazikani, B.; Hopkins, A. L. Nat Rev Drug Discov 2006, 5, 993. (6) Dauter, Z.; Lamzin, V. S.; Wilson, K. S. Current Opinion in Structural Biology 1997, 7, 681. (7) Hansen, C.; Quake, S. R. Current Opinion in Structural Biology 2003, 13, 538. (8) Govada, L.; Carpenter, L.; da Fonseca, P. C. A.; Helliwell, J. R.; Rizkallah, P.; Flashman, E.; Chayen, N. E.; Redwood, C.; Squire, J. M. J Mol Biol 2008, 378, 387. (9) Hansen, C. L.; Skordalakes, E.; Berger, J. M.; Quake, S. R. P Natl Acad Sci USA 2002, 99, 16531. (10) Leng, J.; Salmon, J.-B. Lab Chip 2009, 9, 24. (11) Zheng, B.; Gerdts, C. J.; Ismagilov, R. F. Current Opinion in Structural Biology 2005, 15, 548. (12) Lorber, B.; Delucas, L. J.; Bishop, J. B. J Cryst Growth 1991, 110, 103. (13) Talreja, S.; Perry, S. L.; Guha, S.; Bhamidi, V.; Zukoski, C. F.; Kenis, P. J. A. The Journal of Physical Chemistry B 2010, 114, 4432. (14) Chayen, N. E. Current Opinion in Structural Biology 2004, 14, 577. (15) He, G. W.; Bhamidi, V.; Tan, R. B. H.; Kenis, P. J. A.; Zukoski, C. F. Cryst Growth Des 2006, 6, 1175. (16) Zheng, B.; Tice, J. D.; Roach, L. S.; Ismagilov, R. F. Angew Chem Int Edit 2004, 43, 2508. (17) Li, L.; Mustafi, D.; Fu, Q.; Tereshko, V.; Chen, D. L. L.; Tice, J. D.; Ismagilov, R. F. P Natl Acad Sci USA 2006, 103, 19243. (18) Song, H.; Chen, D. L.; Ismagilov, R. F. Angew Chem Int Edit 2006, 45, 7336. (19) van der Woerd, M.; Ferree, D.; Pusey, M. Journal of Structural Biology 2003, 142, 180. (20) Ng, J. D.; Gavira, J. A.; Garcia-Ruiz, J. M. Journal of Structural Biology 2003, 142, 218. (21) Talreja, S.; Kenis, P. J. A.; Zukoski, C. F. Langmuir 2007, 23, 4516. (22) Hansen, C. L.; Quake, S. R.; Berger, J. M. US, 2007. (23) Newman, J.; Fazio, V. J.; Lawson, B.; Peat, T. S. Cryst Growth Des 2010, 10, 2785. (24) Newman, J.; Xu, J.; Willis, M. C. Acta Crystallographica Section D 2007, 63, 826. (25) Collingsworth, P. D.; Bray, T. L.; Christopher, G. K. J Cryst Growth 2000, 219, 283. (26) Durbin, S. D.; Feher, G. Annu Rev Phys Chem 1996, 47, 171. (27) Talreja, S.; Kim, D. Y.; Mirarefi, A. Y.; Zukoski, C. F.; Kenis, P. J. A. J Appl Crystallogr 2005, 38, 988. (28) Yoshizaki, I.; Nakamura, H.; Sato, T.; Igarashi, N.; Komatsu, H.; Yoda, S. J Cryst Growth 2002, 237, 295. (29) Anderson, M. J.; Hansen, C. L.; Quake, S. R. P Natl Acad Sci USA 2006, 103, 16746. (30) Hansen, C. L.; Sommer, M. O. A.; Quake, S. R. P Natl Acad Sci USA 2004, 101, 14431. (31) Lounaci, M.; Rigolet, P.; Abraham, C.; Le Berre, M.; Chen, Y. Microelectron Eng 2007, 84, 1758. (32) Zheng, B.; Roach, L. S.; Ismagilov, R. F. J Am Chem Soc 2003, 125, 11170. (33) Zhou, X.; Lau, L.; Lam, W. W. L.; Au, S. W. N.; Zheng, B. Anal. Chem. 2007. (34) Cherezov, V.; Caffrey, M. J Appl Crystallogr 2003, 36, 1372. (35) Qutub, Y.; Reviakine, I.; Maxwell, C.; Navarro, J.; Landau, E. M.; Vekilov, P. G. J Mol Biol 2004, 343, 1243. (36) Rummel, G.; Hardmeyer, A.; Widmer, C.; Chiu, M. L.; Nollert, P.; Locher, K. P.; Pedruzzi, I.; Landau, E. M.; Rosenbusch, J. P. Journal of Structural Biology 1998, 121, 82. (37) Gavira, J. A.; Toh, D.; Lopez-Jaramillo, J.; Garcia-Ruiz, J. M.; Ng, J. D. Acta Crystallogr D 2002, 58, 1147. (38) Stevens, R. C. Current Opinion in Structural Biology 2000, 10, 558. (39) Baker, M. Nat Methods 2010, 7, 429. (40) McPherson, A. In Current Topics in Membranes, Volume 63; Volume 63 ed.; DeLucas, L., Ed.; Academic Press: 2009, p 5. (41) Gabrielsen, M.; Gardiner, A. T.; Fromme, P.; Cogdell, R. J. In Current Topics in Membranes, Volume 63; Volume 63 ed.; DeLucas, L., Ed.; Academic Press: 2009, p 127. (42) Page, R. In Methods in Molecular Biology: Structural Proteomics - High Throughput Methods; Kobe, B., Guss, M., Huber, T., Eds.; Humana Press: Totowa, NJ, 2008; Vol. 426, p 345. (43) Caffrey, M. Ann Rev Biophys 2009, 38, 29. (44) Doerr, A. Nat Methods 2006, 3, 244. (45) Brostromer, E.; Nan, J.; Li, L.-F.; Su, X.-D. Biochemical and Biophysical Research Communications 2009, 386, 634. (46) Li, G.; Chen, Q.; Li, J.; Hu, X.; Zhao, J. Anal Chem 2010, 82, 4362. (47) Jia, Y.; Liu, X.-Y. The Journal of Physical Chemistry B 2006, 110, 6949. (48) RCSB Protein Data Bank. http://www.rcsb.org/ (July 11, 2010). (49) Membrane Proteins of Known 3D Structure. http://blanco.biomol.uci.edu/Membrane_Proteins_xtal.html (July 11, 2010). (50) Michel, H. Trends Biochem Sci 1983, 8, 56. (51) Rosenbusch, J. P. Journal of Structural Biology 1990, 104, 134. (52) Garavito, R. M.; Picot, D. Methods 1990, 1, 57. (53) Kulkarni, C. V. 2010; Vol. 12, p 237. (54) Landau, E. M.; Rosenbusch, J. P. P Natl Acad Sci USA 1996, 93, 14532. (55) Pebay-Peyroula, E.; Rummel, G.; Rosenbusch, J. P.; Landau, E. M. Science 1997, 277, 1676. (56) Cherezov, V.; Liu, W.; Derrick, J. P.; Luan, B.; Aksimentiev, A.; Katritch, V.; Caffrey, M. Proteins: Structure, Function, and Bioinformatics 2008, 71, 24. (57) Cherezov, V.; Rosenbaum, D. M.; Hanson, M. A.; Rasmussen, S. G. F.; Thian, F. S.; Kobilka, T. S.; Choi, H. J.; Kuhn, P.; Weis, W. I.; Kobilka, B. K.; Stevens, R. C. Science 2007, 318, 1258. (58) Cherezov, V.; Yamashita, E.; Liu, W.; Zhalnina, M.; Cramer, W. A.; Caffrey, M. J Mol Biol 2006, 364, 716. (59) Jaakola, V. P.; Griffith, M. T.; Hanson, M. A.; Cherezov, V.; Chien, E. Y. T.; Lane, J. R.; IJzerman, A. P.; Stevens, R. C. Science 2008, 322, 1211. (60) Rosenbaum, D. M.; Cherezov, V.; Hanson, M. A.; Rasmussen, S. G. F.; Thian, F. S.; Kobilka, T. S.; Choi, H. J.; Yao, X. J.; Weis, W. I.; Stevens, R. C.; Kobilka, B. K. Science 2007, 318, 1266. (61) Wacker, D.; Fenalti, G.; Brown, M. A.; Katritch, V.; Abagyan, R.; Cherezov, V.; Stevens, R. C. J Am Chem Soc 2010, 132, 11443. (62) Höfer, N.; Aragão, D.; Caffrey, M. Biophys J 2010, 99, L23. (63) Li, L.; Ismagilov, R. F. Ann Rev Biophys 2010. (64) Pal, R.; Yang, M.; Lin, R.; Johnson, B. N.; Srivastava, N.; Razzacki, S. Z.; Chomistek, K. J.; Heldsinger, D. C.; Haque, R. M.; Ugaz, V. M.; Thwar, P. K.; Chen, Z.; Alfano, K.; Yim, M. B.; Krishnan, M.; Fuller, A. O.; Larson, R. G.; Burke, D. T.; Burns, M. A. Lab Chip 2005, 5, 1024. (65) Jayashree, R. S.; Gancs, L.; Choban, E. R.; Primak, A.; Natarajan, D.; Markoski, L. J.; Kenis, P. J. A. J Am Chem Soc 2005, 127, 16758. (66) Wootton, R. C. R.; deMello, A. J. Chem Commun 2004, 266. (67) McPherson, A. J Appl Crystallogr 2000, 33, 397.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Studies of chemokine receptors (CKR) in natural killer- (NK-) cells have already been published, but only a few gave detailed information on its differential expression on blood NK-cell subsets. We report on the expression of the inflammatory and homeostatic CKR on normal blood CD56(+low) CD16(+) and CD56(+high)  CD16(-/+low) NK-cells. Conventional CD56(+low) and CD56(+high) NK-cells present in the normal PB do express CKR for inflammatory cytokines, although with different patterns CD56(+low) NK-cells are mainly CXCR1/CXCR2(+) and CXCR3/CCR5(-/+), whereas mostly CD56(+high) NK-cells are CXCR1/CXCR2(-) and CXCR3/CCR5(+). Both NK-cell subsets have variable CXCR4 expression and are CCR4(-) and CCR6(-). The CKR repertoire of the CD56(+low) NK-cells approaches to that of neutrophils, whereas the CKR repertoire of the CD56(+high) NK-cells mimics that of Th1(+) T cells, suggesting that these cells are prepared to migrate into inflamed tissues at different phases of the immune response. In addition, we describe a subpopulation of NK-cells with intermediate levels of CD56 expression, which we named CD56(+int) NK-cells. These NK-cells are CXCR3/CCR5(+), they have intermediate levels of expression of CD16, CD62L, CD94, and CD122, and they are CD57(-) and CD158a(-). In view of their phenotypic features, we hypothesize that they correspond to a transitional stage, between the well-known CD56(+high) and CD56(+low) NK-cells populations.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Kafirin microparticles have been proposed as an oral nutraceutical and drug delivery system. This study investigates microparticles formed with kafirin extracted from white and raw versus cooked red sorghum grains as an oral delivery system. Targeted delivery to the colon would be beneficial for medication such as prednisolone, which is used in the management of inflammatory bowel disease. Therefore, prednisolone was loaded into microparticles of kafirin from the different sources using phase separation. Differences were observed in the protein content, in vitro protein digestibility, and protein electrophoretic profile of the various sources of sorghum grains, kafirin extracts, and kafirin microparticles. For all of the formulations, the majority of the loaded prednisolone was not released in in vitro conditions simulating the upper gastrointestinal tract, indicating that most of the encapsulated drug could reach the target area of the lower gastrointestinal tract. This suggests that these kafirin microparticles may have potential as a colon-targeted nutraceutical and drug delivery system.