Un modèle animal simple pour l'apprentissage des techniques de microanastomoses vasculaires de congruences diefférentes

BACKGROUND Since the pioneering work of Jacobson and Suarez, microsurgery has steadily progressed and is now used in all surgical specialities, particularly in plastic surgery. Before performing clinical procedures it is necessary to learn the basic techniques in the laboratory. OBJECTIVE To assess an animal model, thereby circumventing the following issues: ethical rules, cost, anesthesia and training time. METHODS Between July 2012 and September 2012, 182 earthworms were used for 150 microsurgical trainings to simulate discrepancy microanastomoses. Training was undertaken over 10 weekly periods. Each training session included 15 simulations of microanastomoses performed using the Harashina technique (earthworm diameters >1.5 mm [n=5], between 1.0 mm and 1.5 mm [n=5], and <1.0 mm [n=5]). The technique is presented and documented. A linear model with main variable as the number of the week (as a numeric covariate) and the size of the animal (as a factor) was used to determine the trend in time of anastomosis over subsequent weeks as well as differences between the different size groups. RESULTS The linear model showed a significant trend (P<0.001) in time of anastomosis in the course of the training, as well as significant differences (P<0.001) between the groups of animal of different sizes. For diameter >1.5 mm, mean anastomosis time decreased from 19.6±1.9 min to 12.6±0.7 min between the first and last week of training. For training involving smaller diameters, the results showed a reduction in execution time of 36.1% (P<0.01) (diameter between 1.0 mm and 1.5 mm) and 40.6% (P<0.01) (diameter <1.0 mm) between the first and last weeks. The study demonstrates an improvement in the dexterity and speed of nodes' execution. CONCLUSION The earthworm appears to be a reliable experimental model for microsurgical training of discrepancy microanastomoses. Its numerous advantages, as discussed in the present report, show that this model of training will significantly grow and develop in the near future.

Age-Independent Cartilage Generation for Synovium-Based Autologous Chondrocyte Implantation.

The articular cartilage layer of synovial joints is commonly lesioned by trauma or by a degenerative joint disease. Attempts to repair the damage frequently involve the performance of autologous chondrocyte implantation (ACI). Healthy cartilage must be first removed from the joint, and then, on a separate occasion, following the isolation of the chondrocytes and their expansion in vitro, implanted within the lesion. The disadvantages of this therapeutic approach include the destruction of healthy cartilage-which may predispose the joint to osteoarthritic degeneration-the necessarily restricted availability of healthy tissue, the limited proliferative capacity of the donor cells-which declines with age-and the need for two surgical interventions. We postulated that it should be possible to induce synovial stem cells, which are characterized by high, age-independent, proliferative and chondrogenic differentiation capacities, to lay down cartilage within the outer juxtasynovial space after the transcutaneous implantation of a carrier bearing BMP-2 in a slow-release system. The chondrocytes could be isolated on-site and immediately used for ACI. To test this hypothesis, Chinchilla rabbits were used as an experimental model. A collagenous patch bearing BMP-2 in a slow-delivery vehicle was sutured to the inner face of the synovial membrane. The neoformed tissue was excised 5, 8, 11 and 14 days postimplantation for histological and histomorphometric analyses. Neoformed tissue was observed within the outer juxtasynovial space already on the 5th postimplantation day. It contained connective and adipose tissues, and a central nugget of growing cartilage. Between days 5 and 14, the absolute volume of cartilage increased, attaining a value of 12 mm(3) at the latter juncture. Bone was deposited in measurable quantities from the 11th day onwards, but owing to resorption, the net volume did not exceed 1.5 mm(3) (14th day). The findings confirm our hypothesis. The quantity of neoformed cartilage that is deposited after only 1 week within the outer juxtasynovial space would yield sufficient cells for ACI. Since the BMP-2-bearing patches would be implanted transcutaneously in humans, only one surgical or arthroscopic intervention would be called for. Moreover, most importantly, sufficient numbers of cells could be generated in patients of all ages.

Caracterización y cuantificación de células progenitoras endoteliales de ratas espontáneamente hipertensas alimentadas con fructosa

Objetivo: Examinar cómo se ve afecta la participación de las células progenitoras endoteliales (CPE) por la resistencia a la insulina (IR) asociada a un modelo experimental de síndrome metabólico (SM), generado por la administración crónica de fructosa a ratas espontáneamente hipertensas. Material y métodos: Ratas WKY y SHR, macho, distribuidas en 4 grupos (n=8 c/u): WKY: controles; FFR: WKY recibiendo fructosa en agua de bebida al 10 % (v/v) durante 6 semanas; SHR; FFHR: SHR recibiendo fructosa en agua de bebida al 10 % (v/v) durante 6 semanas. Al finalizar el protocolo se determinó: presión arterial sistólica, variables bioquímicas, índice HOMA, cuantificación por citometría de flujo de los niveles de CPE en sangre periférica y en médula ósea, inmunofluorescencia en cultivo celular, para identificar los marcadores CD34 y VEGFR-2, recuento de colonias de CPE y actividad de NAD(P)H-oxidasa en tejido aórtico. Resultados: Se confirmó el modelo experimental en base a las variables metabólicas analizadas. Se observó una disminución en los niveles de CPE; en sangre periférica y médula ósea, la que se hace más importante en los grupos de animales tratados con fructosa. En estos también hay menor número de colonias de CPE desarrolladas en cultivo celular y presentan un aumento en los niveles de estrés oxidativo, estimado por la actividad de NAD(P)H oxidasa. Conclusión: el SM causado por la administración crónica de fructosa en FFHR ha demostrado generar una disminución en los niveles de CPE, así como en su capacidad funcional. Los mecanismos intracelulares que producen este fenómeno podrían estar desencadenados por el grado de IR que presenta este modelo experimental.

Time Evolution of the Fractal Dimension in Turbulent Plumes

Turbulent mixing is a very important issue in the study of geophysical phenomena because most fluxes arising in geophysics fluids are turbulent. We study turbulent mixing due to convection using a laboratory experimental model with two miscible fluids of different density with an initial top heavy density distribution. The fluids that form the initial unstable stratification are miscible and the turbulence will produce molecular mixing. The denser fluid comes into the lighter fluid layer and it generates several forced plumes which are gravitationally unstable. As the turbulent plumes develop, the denser fluid comes into contact with the lighter fluid layer and the mixing process grows. Their development is caused by the lateral interaction between these plumes at the complex fractal surface between the dense and light fluids

Estudio Energético y de Explotación de diferentes conjuntos de Labranza para el Cultivo de la Yuca (Manihot esculenta, Crantz) en un Fluvisol

El trabajo se desarrolló en tres etapas de investigación, las dos primeras fueron realizadas en la Unidad Básica de Producción Cooperativa “El Palmar” perteneciente a la Empresa de Cultivos Varios "Paquito Rosales Benítez" del municipio de Yara, provincia de Granma, Cuba, el cual se llevó a cabo en el período de abril de 2005 a enero de 2008; y de noviembre de 2012 a febrero de 2013 respectivamente; la tercera se ejecutó de mayo a junio de 2013 en un canal de suelo ubicado en la nave de maquinaria del Departamento de Ingeniería Rural de la Universidad Politécnica de Madrid. En la primera etapa se evaluaron los conjuntos tractor-apero que intervienen en la realización de cuatro sistemas de labranza para el cultivo de la yuca, variedad Enana Rosada. Los sistemas de laboreo fueron: T1, labor de aradura (alzado) con un arado de discos seguidas de un pase de grada de discos un pase cruzado de un escarificador y un pase con una grada de discos (5 labores); T2, laboreo mínimo que comprende una labor profunda con un escarificador con saetas seguido por una labor de grada de discos (2 labores); T3, laboreo mínimo en el que se realizaron dos pases cruzados con escarificador y una labor de fresado con una fresadora de eje horizontal (3 labores) y; T4, dos labores cruzadas con la grada de discos seguidas por otras dos pases cruzados con el escarificador y una última labor con la grada de discos (5 labores). Se realizaron cinco observaciones de la jornada laboral para cada uno de los conjuntos tractorapero de cada sistema de laboreo en tres campañas, evaluando el consumo de combustible, las diferentes productividades, los índices y coeficientes tecnológicos y de explotación, el rendimiento del cultivo el balance energético del mismo, y la emisión de CO2 equivalente. En la segunda etapa se determinó la fuerza de tiro requerida por un órgano de trabajo de un escarificador con saetas en condiciones de campo, en tres profundidades (15, 25 y 35 cm) y a cuatro velocidades de trabajo (1,30; 1,49; 1,65 y 2,32 m s-1). En la tercera etapa se determinaron las fuerzas horizontales y verticales en una maqueta experimental de un órgano de trabajo de un escarificador con saetas (1:5 del tamaño real del apero) en cuatro contenidos de humedad (60, 90, 120 y 150 g kg-1), tres densidades del suelo (1,1; 1,2 y 1,3 Mg m-3) y una velocidad de 40 mm s-1. Se comparan los resultados del apero a tamaño real y la maqueta experimental. Los mejores resultados relacionados con los indicadores evaluados a la maquinaria correspondieron al sistema de laboreo mínimo T2, el cual presenta la mejor productividad por hora de tiempo limpio (1,34 ha h-1) y de tiempo operativo (1,27 ha h-1), con diferencias significativas respecto al resto de los sistemas de labranza evaluados; así como el menor consumo de combustible (32,4 L ha-1) y el mejor comportamiento energético, con diferencias significativas respecto a las demás, resultando inferior en un 63, 30 y 58 % con relación a T1, T3 y T4. La fuerza de tiro del órgano de trabajo del escarificador con saetas se incrementa con la densidad del suelo y con la profundidad, y velocidad de trabajo. Las fuerzas horizontales y verticales de la maqueta se incrementan linealmente con la densidad y humedad del suelo. Existe una relación lineal entre las fuerzas de tiro y horizontales medidas en el apero real y en el apero a escala con el contenido de humedad del suelo. ABSTRACT The work was developed in three stages of research, the first two were conducted in the Basic Unit of Cooperative Production "El Palmar" belonging to the agricultural farm "Paquito Rosales Benitez" in the municipality of Yara, Granma Province, Cuba, the which was conducted in the period from April 2005 to January 2008 and November 2012 and February 2013 respectively, the third from May to June 2013 on a soilbin located in the ship's machinery of Rural Engineering Department of the Technical University of Madrid. In the first stage were evaluated tractor-implement sets involved in the completion of four tillage systems for cassava, Pink Dwarf variety. Tillage systems were: T1, plowing (elevation) with a disk plow followed by a pass of disc harrow, a cross of scarifier and a pass with a disc harrow (5 labours), T2, minimum tillage comprising a deep work with a scarifier with a bolt followed by a disc harrow work (two labours); T3, minimum tillage which two crossed passes performed with scarifier and milling labour with a horizontal axis milling machine (three labours) and, T4, two crossed labours with disc harrow followed by two crossed passes with the scarifier and a final labour with disc harrow (5 labours). Five observations of the working day for each tractor-implement sets of each tillage system in three seasons were performed, evaluating fuel consumption, different productivities, indexes and technological and operational factors, the crop yield energy balance and the emission of CO2 equivalent. In the second stage we determined the draft force required by a working body of a scarifier with bolt under field conditions at three depths (15, 25 and 35 cm) and four operating speeds (1.30, 1.49 , 1.65 and 2.32 m s-1). In the third stage were determined horizontal and vertical forces on an experimental model of a working body of a scarifier with bolt (1:5 real scale of the implement) in four moisture contents (60, 90, 120 and 150 g kg -1), three soil densities (1.1, 1.2, and 1.3 Mg m-3) and a speed of 40 mm s-1.The results of real scale implement the experimental model were compared. The best results related to evaluated machinery indicators corresponded to T2 minimum tillage system, which presents the best productivity per hour clean time (1.34 ha h-1) and operating time (1.27 ha h-1), with significant differences compared to other tillage systems evaluated; as well as lower fuel consumption (32.4 L ha-1) and the best energy performance, with significant differences from the other, resulting lower by 63, 30 and 58% compared to T1, T3 and T4. The draft force of the working body of the scarifier with bolt increases with soil density and depth, and speed of work. The horizontal and vertical forces of the model increase linearly with density and soil moisture. A linear relationship exists between the shot and horizontal forces measured at the real implement and scale implement with the soil moisture content.

Bases para un diseño metodológico de estimulación binaural como terapia acústica en neurología

La medicina y la ingeniería del siglo XXI han dado como fruto numerosos avances para la sociedad aunque en la mayoría de los casos los tratamientos suelen ser costosos e invasivos. La educación que recibe la sociedad sobre la salud es escasa, ya que sólo vamos al médico cuando realmente estamos enfermos. Este trabajo presenta nuestra apuesta por las terapias complementarias, para el desarrollo de una metodología terapéutica no invasiva y con un costo muy bajo. La finalidad de esta Tesis, que se enmarca en un equipo multidisciplinar, fruto de la estrecha colaboración en el que participan psicopedagogos, ingenieros y médicos, es perfilar una metodología que luego pueda ser aplicable a patologías neurológicas. Aquí, dejamos sentadas las bases. Faltarán nuevos investigadores que continúen este camino para tener una base de datos lo suficientemente extensa de registros de sujetos que hayan sido sometidos a terapia binaural, para poder sacar unas conclusiones sólidas. La aportación de esta Tesis deja cubierta la aplicación, selección, procesado de señal y desarrollo de algoritmos, test cognitivos indicados para el caso específico que nos ocupa, cálculo de incertidumbre del sistema utilizado para la aplicación del estímulo y desarrollo de un test psicoacústico específico. EL empleo del sonido en medicina como es la musicoterapia o sonoterapia ha experimentado una gran difusión en los últimos años, más de 100.000 nuevas citas bibliográficas han aparecido con respecto al año anterior. Sin embargo, son escasísimas las que hacen referencia a las características físico acústicas del sonido empleado, tan sólo hemos encontrado una par de ellas que correlacionan las características físicas del sonido con el tipo de respuesta terapéutica. No encontramos citas bibliográficas específicas que planteen un modelo experimental científico capaz de reproducir las mismas respuestas ante los mismos parámetros y estímulos. En esta Tesis proponemos el uso de estimulación sonora binaural, que consiste en la utilización de dos tonos puros idénticos pero ligeramente diferentes en frecuencia que se presentan de manera separada cada uno en un oído, como consecuencia, la persona que recibe la estimulación percibe un tercer tono, llamado tono binaural, formado por la diferencia de frecuencia de ambos variando su amplitud. Existen estudios que sugieren que dichas frecuencias binaurales pueden modificar los patrones eléctricos de la actividad cerebral y los niveles de arousal, conociéndose en la literatura bajo el nombre de “entrainment”. Tras la revisión bibliográfica del estado del arte, podemos concluir que es necesario el desarrollo de estudios doble ciego bien diseñados, con el objetivo de establecer una base sólida sobre los efectos de este tipo de estimulación, ya que la mayoría de los beneficios documentados se refieren a muestras muy pequeñas y con poco rigor científico, siendo los resultados positivos obtenidos debidos al efecto placebo. La tecnología binaural es barata siendo cualquier avance en esta dirección de interés público. El objetivo concreto de la investigación es estudiar el potencial de las ondas binaurales en un área en particular: tareas que requieren atención y concentración. Se busca obtener cualquier cambio en las ondas cerebrales que se puedan correlar con la mejoras. A la vista de los resultados de estas investigaciones se intentará aplicar esta metodología en neuropatologías que presenten alguna deficiencia en el área de atención como es el Trastorno de espectro Autista. En esta Tesis presentamos los resultados de dos estudios independientes, el primero para sentar las bases del método (tiempos, diseño de estimulaciones, procesado) en una muestra de 78 adultos sanos, el segundo a partir de los resultados obtenidos en el primero, afinando la metodología y para un grupo de 20 niños entre 8 y 12 años, los resultados del segundo estudio sirven para justificar su aplicación en niños con TEA que presenten déficit de atención. ABSTRACT Medicine and engineering in the 21st century have resulted in advances for society but in most cases the treatments are often costly and invasive. The health education society receive is scarce, since only go to the doctor when we are really sick. With this work I present my commitment to complementary therapies, my little grain of sand in the development of a noninvasive therapeutic approach and very low cost, well and can be used in a preventive manner resulting in a society with less sick. The purpose of this thesis is to outline a methodology that can then be applied to neurological diseases, here we lay the groundwork. New researchers are needed to continue this path for a sufficiently extensive records database of subjects who have undergone binaural therapy, and so to draw firm conclusions. The contribution of this thesis includes: the application, selection, signal processing and algorithm development, indicated cognitive tests for the specific case at hand, calculation of system uncertainty of the system and development of a specific psychoacoustic test. The use of sound in medicine, such as music therapy or sound therapy has experienced a great diffusion in recent years, more than 100,000 new citations have appeared over the previous year but very few are those referring to acoustic physical characteristics of sound employee, we have only found a couple of them that physical sound characteristics are correlated with the therapeutic response. We found no specific citations posing a scientific experimental model capable of reproducing the same answers to the same parameters and stimuli. In this thesis we propose the use of binaural sound stimulation which involves the use of two identical but slightly different in frequency pure tones presented separately each in one ear, as a result the subject perceives a third tone, called binaural tone, formed by the difference in frequency with amplitude variations Studies suggest that these binaural frequencies can modify the electrical patterns of brain activity and arousal levels, being known in the literature under the name of “entrainment”. After the literature review of the state of the art, we conclude, it is necessary to develop well-designed double-blind studies, in order to establish a solid foundation on the effects of such stimulation, since most of the documented benefits relate to very small samples and unscientific may be obtained positive results due to the placebo effect. The binaural technology is cheap being any progress in this direction in the public interest. The specific objective of the research is to study the potential of binaural waves in a particular area: tasks requiring attention and concentration also we want to get any change in brain waves that can correlate with improvements. In view of the results of this research we seek to apply this methodology in neuropathology presenting any deficiency in the area of attention such as Autism Spectrum Disorder. In this thesis we present the results of two independent studies, the first to lay the foundation of the method (times, stimulation design, processing) in a sample of 78 healthy adults, the second from the results obtained in the first, refine the methodology for a group of 20 children between 8 and 12 years, the results of the second study used to justify its use in children with ASD that present attention deficit.

Compromisos en la adaptación a distintos huéspedes en virus de plantas

Los patógenos han desarrollado estrategias para sobrevivir en su entorno, infectar a sus huéspedes, multiplicarse dentro de estos y posteriormente transmitirse a otros huéspedes. Todos estos componentes hacen parte de la eficacia biológica de los patógenos, y les permiten ser los causantes de enfermedades infecciosas tanto en hombres y animales, como en plantas. El proceso de infección produce efectos negativos en la eficacia biológica del huésped y la gravedad de los efectos, dependerá de la virulencia del patógeno. Por su parte, el huésped ha desarrollado mecanismos de respuesta en contra del patógeno, tales como la resistencia, por la que reduce la multiplicación del patógeno, o la tolerancia, por la que disminuye el efecto negativo de la infección. Estas respuestas del huésped a la infección producen efectos negativos en la eficacia biológica del patógeno, actuando como una presión selectiva sobre su población. Si la presión selectiva sobre el patógeno varía según el huésped, se predice que un mismo patógeno no podrá aumentar su eficacia biológica en distintos huéspedes y estará más adaptado a un huésped y menos a otro, disminuyendo su gama de huéspedes. Esto supone que la adaptación de un patógeno a distintos huéspedes estará a menudo dificultada por compromisos (trade-off) en diferentes componentes de la eficacia biológica del patógeno. Hasta el momento, la evidencia de compromisos de la adaptación del patógeno a distintos huéspedes no es muy abundante, en lo que se respecta a los virus de plantas. En las últimas décadas, se ha descrito un aumento en la incidencia de virus nuevos o previamente descritos que producen enfermedades infecciosas con mayor gravedad y/o diferente patogenicidad, como la infección de huéspedes previamente resistentes. Esto se conoce como la emergencia de enfermedades infecciosas y está causada por patógenos emergentes, que proceden de un huésped reservorio donde se encuentran adaptados. Los huéspedes que actúan como reservorios pueden ser plantas silvestres, que a menudo presentan pocos síntomas o muy leves a pesar de estar infectados con diferentes virus, y asimismo se encuentran en ecosistemas con ninguna o poca intervención humana. El estudio de los factores ecológicos y biológicos que actúan en el proceso de la emergencia de enfermedades infecciosas, ayudará a entender sus causas para crear estrategias de prevención y control. Los virus son los principales patógenos causales de la emergencia de enfermedades infecciosas en humanos, animales y plantas y un buen modelo para entender los procesos de la emergencia. Asimismo, las plantas a diferencia de los animales, son huéspedes fáciles de manipular y los virus que las afectan, más seguros para el trabajo en laboratorio que los virus de humanos y animales, otros modelos también usados en la investigación. Por lo tanto, la interacción virus – planta es un buen modelo experimental para el estudio de la emergencia de enfermedades infecciosas. El estudio de la emergencia de virus en plantas tiene también un interés particular, debido a que los virus pueden ocasionar pérdidas económicas en los cultivos agrícolas y poner en riesgo la durabilidad de la resistencia de plantas mejoradas, lo que supone un riesgo en la seguridad alimentaria con impactos importantes en la sociedad, comparables con las enfermedades infecciosas de humanos y animales domésticos. Para que un virus se convierta en un patógeno emergente debe primero saltar desde su huésped reservorio a un nuevo huésped, segundo adaptarse al nuevo huésped hasta que la infección dentro de la población de éste se vuelva independiente del reservorio y finalmente debe cambiar su epidemiología. En este estudio, se escogió la emergencia del virus del mosaico del pepino dulce (PepMV) en el tomate, como modelo experimental para estudiar la emergencia de un virus en una nueva especie de huésped, así como las infecciones de distintos genotipos del virus del moteado atenuado del pimiento (PMMoV) en pimiento, para estudiar la emergencia de un virus que aumenta su patogenicidad en un huésped previamente resistente. El estudio de ambos patosistemas nos permitió ampliar el conocimiento sobre los factores ecológicos y evolutivos en las dos primeras fases de la emergencia de enfermedades virales en plantas. El PepMV es un patógeno emergente en cultivos de tomate (Solanum lycopersicum) a nivel mundial, que se describió primero en 1980 infectando pepino dulce (Solanum muricatum L.) en Perú, y casi una década después causando una epidemia en cultivos de tomate en Holanda. La introducción a Europa posiblemente fue a través de semillas infectadas de tomate procedentes de Perú, y desde entonces se han descrito nuevos aislados que se agrupan en cuatro cepas (EU, LP, CH2, US1) que infectan a tomate. Sin embargo, el proceso de su emergencia desde pepino dulce hasta tomate es un interrogante de gran interés, porque es uno de los virus emergentes más recientes y de gran importancia económica. Para la emergencia de PepMV en tomate, se recolectaron muestras de tomate silvestre procedentes del sur de Perú, se analizó la presencia y diversidad de aislados de PepMV y se caracterizaron tanto biológicamente (gama de huéspedes), como genéticamente (secuencias genomicas). Se han descrito en diferentes regiones del mundo aislados de PMMoV que han adquirido la capacidad de infectar variedades previamente resistentes de pimiento (Capsicum spp), es decir, un típico caso de emergencia de virus que implica la ampliación de su gama de huéspedes y un aumento de patogenicidad. Esto tiene gran interés, ya que compromete el uso de variedades resistentes obtenidas por mejora genética, que es la forma de control de virus más eficaz que existe. Para estudiar la emergencia de genotipos altamente patogénicos de PMMoV, se analizaron clones biológicos de PMMoV procedentes de aislados de campo cuya patogenicidad era conocida (P1,2) y por mutagénesis se les aumentó la patogenicidad (P1,2,3 y P1,2,3,4), introduciendo las mutaciones descritas como responsables de estos fenotipos. Se analizó si el aumento de la patogenicidad conlleva un compromiso en la eficacia biológica de los genotipos de PMMoV. Para ello se evaluaron diferentes componentes de la eficacia biológica del virus en diferentes huéspedes con distintos alelos de resistencia. Los resultados de esta tesis demuestran: i). El potencial de las plantas silvestres como reservorios de virus emergentes, en este caso tomates silvestres del sur de Perú, así como la existencia en estas plantas de aislados de PepMV de una nueva cepa no descrita que llamamos PES. ii) El aumento de la gama de huéspedes no es una condición estricta para la emergencia de los virus de plantas. iii) La adaptación es el mecanismo más probable en la emergencia de PepMV en tomate cultivado. iv) El aumento de la patogenicidad tiene un efecto pleiotrópico en distintos componentes de la eficacia biológica, así mismo el signo y magnitud de este efecto dependerá del genotipo del virus, del huésped y de la interacción de estos factores. ABSTRACT host Pathogens have evolved strategies to survive in their environment, infecting their hosts, multiplying inside them and being transmitted to other hosts. All of these components form part of the pathogen fitness, and allow them to be the cause of infectious diseases in humans, animals, and plants. The infection process produces negative effects on the host fitness and the effects severity will depend on the pathogen virulence. On the other hand, hosts have developed response mechanisms against pathogens such as resistance, which reduces the growth of pathogens, or tolerance, which decreases the negative effects of infection. T he se responses of s to infection cause negative effects on the pathogen fitness, acting as a selective pressure on its population. If the selective pressures on pathogens va ry according to the host s , probably one pathogen cannot increase its fitness in different hosts and will be more adapted to one host and less to another, decreasing its host range. This means that the adaptation of one pathogen to different hosts , will be often limited by different trade - off components of biological effectiveness of pathogen. Nowadays , trade - off evidence of pathogen adaptation to different hosts is not extensive, in relation with plant viruses. In last decades, an increase in the incidence of new or previously detected viruses has been described, causing infectious diseases with increased severity and/or different pathogenicity, such as the hosts infection previously resistants. This is known as the emergence of infectious diseases and is caused by emerging pathogens that come from a reservoir host where they are adapted. The hosts which act as reservoirs can be wild plants, that often have few symptoms or very mild , despite of being infected with different viruses, and being found in ecosystems with little or any human intervention. The study of ecological and biological factors , acting in the process of the infectious diseases emergence will help to understand its causes to create strategies for its prevention and control. Viruses are the main causative pathogens of the infectious diseases emergence in humans, animals and plants, and a good model to understand the emergency processes. Likewise, plants in contrast to animals are easy host to handle and viruses that affect them, safer for laboratory work than viruses of humans and animals, another models used in research. Therefore, the interaction plant-virus is a good experimental model for the study of the infectious diseases emergence. The study of virus emergence in plants also has a particular interest, because the viruses can cause economic losses in agricultural crops and threaten the resistance durability of improved plants, it suppose a risk for food security with significant impacts on society, comparable with infectious diseases of humans and domestic animals. To become an emerging pathogen, a virus must jump first from its reservoir host to a new host, then adapt to a new host until the infection within the population becomes independent from the reservoir, and finally must change its epidemiology. In this study, the emergence of pepino mosaic virus (PepMV) in tomato, was selected as experimental model to study the emergence of a virus in a new host specie, as well as the infections of different genotypes of pepper mild mottle virus (PMMoV) in pepper, to study the emergence of a virus that increases its pathogenicity in a previously resistant host. The study of both Pathosystems increased our knowledge about the ecological and evolutionary factors in the two first phases of the emergence of viral diseases in plants. The PepMV is an emerging pathogen in tomato (Solanum lycopersicum L.) in the world, which was first described in 1980 by infecting pepino (Solanum muricatum L.) in Peru, and almost after a decade caused an epidemic in tomato crops in Netherlands. The introduction to Europe was possibly through infected tomato seeds from Peru, and from then have been described new isolates that are grouped in four strains (EU, LP, CH2, US1) that infect tomato. However, the process of its emergence from pepino up tomato is a very interesting question, because it is one of the newest emerging viruses and economically important. For the PepMV emergence in tomato, wild tomato samples from southern Peru were collected, and the presence and diversity of PepMV isolates were analyzed and characterized at biological (host range) and genetics (genomic sequences) levels. Isolates from PMMoV have been described in different world regions which have acquired the ability to infect pepper varieties that were previously resistants (Capsicum spp), it means, a typical case of virus emergence which involves the host range extension and an increased pathogenicity. This is of great interest due to involve the use of resistant varieties obtained by breeding, which is the most effective way to control virus. To study the emergence of highly pathogenic genotypes of PMMoV, biological clones from field isolates whose pathogenicity was known were analyzed (P1,2) and by mutagenesis we increased its pathogenicity (P1,2,3 and P1,2, 3,4), introducing the mutations described as responsible for these phenotypes. We analyzed whether the increased pathogenicity involves a trade-off in fitness of PMMoV genotypes. For this aim, different components of virus fitness in different hosts with several resistance alleles were evaluated. The results of this thesis show: i). The potential of wild plants as reservoirs of emerging viruses, in this case wild tomatoes in southern Peru, and the existence in these plants of PepMV isolates of a new undescribed strain that we call PES. ii) The host range expansion is not a strict condition for the plant virus emergence. iii) The adaptation is the most likely mechanism in the PepMV emergence in cultivated tomato. iv) The increased pathogenicity has a pleiotropic effect on several fitness components, besides the sign and magnitude of this effect depends on the virus genotype, the host and the interaction of both.

Análisis del rendimiento académico del alumnado de último curso de ingeniería mediante nuevas tecnologías en el proceso de enseñanza‐aprendizaje

En el marco del Espacio Europeo de Educación Superior los estudios deben de estar orientados a facilitar la movilidad de los futuros egresados para que su inclusión en mundo laboral sea global y en este contexto, los sistemas educativos deben introducir cambios en el proceso de enseñanza – aprendizaje y en la gestión. Como uno de los elementos básicos de la creación del EEES es el aprendizaje a lo largo de la vida, se deben adoptar metodologías que doten al estudiante de capacidades para poder enfrentarse a todos los retos de la vida laboral. A través de la presente investigación se trata de aportar una visión real de la aplicación efectiva de un modelo de Aprendizaje Basado en Problemas y el Método del Caso acompañados de una fuerte Acción Tutorial y el uso de la Tecnología de la Información y Comunicación en la Universidad (TIC). Se considera asimismo de gran interés para la mejora del aprendizaje conocer cuál es la opinión real de los estudiantes universitarios, ya que son los principales implicados en relación a un modelo formativo apoyado en el Aprendizaje Basado en Problemas, el Método del Caso, la Acción Tutorial y utilización de las TIC. A través de este estudio se pretende comprobar y valorar cual es la visión real que los alumnos tienen de estas aplicaciones y como las utilizan. Para ello durante los últimos cursos se ha trabajado con alumnos de los últimos cursos de la Escuela Técnica Superior de Ingeniería Civil de la Universidad Politécnica de Madrid aplicando técnicas de Aprendizaje Basado en Problemas y el Método del Caso conjuntamente con la utilización de un Modelo de Acción Tutorial y el uso de la TIC. La tarea se ha centrado en desarrollar, a lo largo de los cursos 2009-10, 2010-11, 2011-12 y 2013-14, un modelo de Acción Tutorial con los alumnos matriculados en las asignaturas de Caminos I, Caminos II, Aforos y Ordenación del Tráfico, asignaturas de la titulación de Ingeniería Técnica de Obras Públicas, y por otro lado, en la asignatura de Caminos, perteneciente a la titulación de Ingeniería Civil. Mediante cuestionarios al inicio y final del curso, se ha conocido cuál es la opinión que poseen los alumnos sobre esta acción. A continuación, durante los cursos 2010-11, 2011-12 y 2013-14 se desarrolla un modelo experimental para evaluar las mejoras, tanto de rendimiento como de adquisición de competencias, utilizando el Aprendizaje Basado en Problemas y el Método del Caso acompañados de las TIC en el proceso de enseñanza–aprendizaje como modelo de Acción Tutorial con alumnos. ABSTRACT Abstract In the frame of the European Higher Education Area, the studies must be faced to facilitate mobility of future graduates for inclusion in the workplace is global and in this context, educational systems must introduce changes in the process of education-learning and management. Since one of the basic elements of the creation of the EHEA is learning throughout life, there must be adopted methodologies that provide the student of aptitudes to be able to face all the challenges of the labor life. Through this research it is provided a real vision of the effective application of a Model of Learning Based on Problems and the Case Method accompanied by a strong Tutorial Action and the use of ITC in the University. It is also considered of great interest for the improvement of learning to know what the real opinion of the college students is, as they are the main players in relation in a training model based on Problem-Based Learning, the Case Method, the Tutorial Action and Use of ICT. Through this study it is expected to verify and assess which is the real vision that students have about these applications and how they use them. In order to achieve the goal of this research project, during the last three years I have been working with students of last courses of the Civil Engineering School of the Technical University of Madrid applying with them techniques of Problem-Based Learning and the Case Method together with the use of a Model Action Tutorial and the Use of Information Technology and Communication (ICT). The task has focused on developing, over the 2009-10, 2010-11, 2011-12 and 2013-14 courses, a model of Tutorial Action with students enrolled in the subjects of Roads I, Roads II, Traffic Gauging and Traffic Management, all of them of the old degree in Civil Engineering (1971 Study Plan), and secondly, on the subject of Roads which belong to the current degree of Civil Engineering. Using questionnaires at the beginning and end of the course the perception that students have on this action.. Then, during the 2010-11, 2011-12 and 2013-14 courses an experimental model is developed to evaluate improvements in both performance and skills acquisition, using Problem-Based Learning and the Case Method together with the ICT in Teaching-Learning Pprocess as a model of Tutorial Action with students.

Caracterización geométrica y lumínica de los atrios incorporados en los edificios de Santiago de Chile

El atrio incorporado en los edificios ha sido un recurso espacial que tempranamente se difundió a nivel global, siendo adoptado por las distintas arquitecturas locales en gran parte del mundo. Su masificación estuvo favorecida primero por la rápida evolución de la tecnología del acero y el vidrio, a partir del siglo XIX, y en segundo termino por el posterior desarrollo del hormigón armado. Otro aspecto que explica tal aceptación en la arquitectura contemporánea, es de orden social y radica en la llamativa cavidad del espacio describiendo grandes dimensiones y favoreciendo con ello, el desarrollo de una multiplicidad de usos en su interior que antes eran impensados. Al interior del atrio, la luz natural es clave en las múltiples vivencias que alberga y sea tal vez la condición ambiental más valorada, ya que entrega una sensación de bienestar al conectarnos visualmente con el ambiente natural. Por esta razón de acuerdo al método hipotético deductivo, se evaluaron los efectos de la configuración geométrica, la cubierta y la orientación en el desempeño de la iluminación natural en la planta baja, a partir un modelo extraído desde el inventario de los edificios atrio construidos en Santiago de Chile, en los últimos 30 años que fue desarrollado en el capitulo 2. El análisis cuantitativo de los edificios inventariados se elaboró en el capítulo 3, considerando las dimensiones de los atrios. Simultáneamente fueron clasificados los aspectos constructivos, los materiales y las características del ambiente interior de cada edificio. En esta etapa además, fueron identificadas las variables de estudio de las proporciones geométricas de la cavidad del atrio con los coeficientes de aspecto de las proporciones, en planta (PAR), en corte (SAR) y de la cavidad según (WI), (AR) y (RI). Del análisis de todos estos parámetros se extrajo el modelo de prueba. El enfoque del estudio del capítulo 4 fue la iluminación natural, se revisaron los conceptos y el comportamiento en el atrio, a partir de un modelo físico construido a escala para registro de la iluminancia bajo cielo soleado de la ciudad. Más adelante se construyó el modelo en ambiente virtual, relacionando las variables determinadas por la geometría de la cavidad y el cerramiento superior; examinándose de esta manera distintas transparencias, proporciones de apertura, en definitiva se evaluó un progresivo cerramiento de las aberturas, verificando el ingreso de la luz y disponibilidad a nivel de piso con la finalidad, de proveer lineamientos útiles en una primera etapa del diseño arquitectónico. Para el análisis de la iluminación natural se revisaron diferentes métodos de cálculo con el propósito de evaluar los niveles de iluminancia en un plano horizontal al interior del atrio. El primero de ellos fue el Factor de Luz Día (FLD) que corresponde, a la proporción de la iluminancia en un punto de evaluación interior respecto, la cantidad proveniente del exterior bajo cielo nublado, a partir de la cual se obtuvo resultados que revelaron la alta luminosidad del cielo nublado de la ciudad. Además fueron evaluadas las recientes métricas dinámicas que dan cuenta, de la cantidad de horas en las cuales de acuerdo a los extensos registros meteorológico de la ciudad, permitieron obtener el porcentajes de horas dentro de las cuales se cumplió el estándar de iluminancia requerido, llamado autonomía lumínica (DA) o mejor aún se permanece dentro de un rango de comodidad visual en el interior del atrio referido a la iluminancia diurna útil (UDI). En el Capítulo 5 se exponen los criterios aplicados al modelo de estudio y cada una de las variantes de análisis, además se profundizó en los antecedentes y procedencia de las fuentes de los registros climáticos utilizados en las simulaciones llevadas a cabo en el programa Daysim operado por Radiance. Que permitieron evaluar el desempeño lumínico y la precisión, de cada uno de los resultados para comprobar la disponibilidad de iluminación natural a través de una matriz. En una etapa posterior se discutieron los resultados, mediante la comparación de los datos logrados según cada una de las metodologías de simulación aplicada. Finalmente se expusieron las conclusiones y futuras lineas de trabajo, las primeras respecto el dominio del atrio de cuatro caras, la incidencia del control de cerramiento de la cubierta y la relación establecida con la altura; indicando en lo específico que las mediciones de iluminancia bajo el cielo soleado de verano, permitieron aclarar, el uso de la herramienta de simulación y método basado en el clima local, que debido a su reciente desarrollo, orienta a futuras líneas de trabajo profundizando en la evaluación dinámica de la iluminancia contrastado con monitorización de casos. ABSTRACT Atriums incorporated into buildings have been a spatial resource that quickly spread throughout the globe, being adopted by several local architecture methods in several places. Their widespread increase was highly favored, in the first place, with the rapid evolution of steel and glass technologies since the nineteen century, and, in second place, by the following development of reinforced concrete. Another issue that explains this success into contemporary architecture is associated with the social approach, and it resides in the impressive cavity that describes vast dimensions, allowing the development of multiple uses in its interior that had never been considered before. Inside the atrium, daylight it is a key element in the many experiences that involves and it is possibly the most relevant environmental factor, since it radiates a feeling of well-being by uniting us visually with the natural environment. It is because of this reason that, following the hypothetical deductive method, the effects in the performance of daylight on the floor plan were evaluated considering the geometric configuration, the deck and orientation factors. This study was based in a model withdrawn from the inventory of atrium buildings that were constructed in Santiago de Chile during the past thirty years, which will be explained later in chapter 2. The quantitative analysis of the inventory of those buildings was elaborated in chapter 3, considering the dimensions of the atriums. Simultaneously, several features such as construction aspects, materials and environmental qualities were identified inside of each building. At this stage, it were identified the variables of the geometric proportions of the atrium’s cavity with the plan aspect ratio of proportions in first plan (PAR), in section (SAR) and cavity according to well index (WI), aspect ratio (AR) and room index (RI). An experimental model was obtained from the analysis of all the mentioned parameters. The main focus of the study developed in chapter 4 is daylight. The atrium’s concept and behavior were analyzed from a physical model built under scale to register the illuminances under clear, sunny sky of the city. Later on, this physical model was built in a virtual environment, connecting the variables determined by the geometry of the cavity and the superior enclosure, allowing the examination of transparencies and opening proportions. To summarize, this stage consisted on evaluating a progressive enclosure of the openings, checking the access of natural light and its availability at the atrium floor, in an effort to provide useful guidelines during the first stage of the architectural design. For the analysis of natural lighting, several calculations methods were used in order to determine the levels of illuminances in a horizontal plane inside of the atrium. The first of these methods is the Daylight Factor (DF), which consists in the proportion of light in an evaluation interior place with the amount of light coming from the outside in a cloudy day. Results determined that the cloudy sky of the city has high levels of luminosity. In addition, the recent dynamic metrics were evaluated which reflects the hours quantity. According to the meteorological records of the city’s climate, the standard of illuminance- a standard measure called Daylight Autonomy (DA) – was met. This is even better when the results stay in the line of visual convenience within the atrium, which is referred to as Useful Daylight Illuminance (UDI). In chapter 5, it was presented the criteria applied to the study model and on each of the variants of the analysis. Moreover, the information of the climate records used for the simulations - carried out in the Daysim program managed by Radiance – are detailed. These simulations allowed the observation of the daylight performance and the accuracy of each of the results to confirm the availability of natural light through a matrix. In a later stage, the results were discussed comparing the collected data in each of the methods of simulation used. Finally, conclusions and further discussion are presented. Overall, the four side atrium’s domain and the effect of the control of the cover’s enclosure. Specifically, the measurements of the daylight under summer’s clear, sunny sky allowing clarifying the use of the simulation tool and the method based on the local climate. This method allows defining new and future lines of work deepening on the dynamic of the light in contrast with the monitoring of the cases.

The solution structure of the N-terminal domain of α2-macroglobulin receptor-associated protein

The three-dimensional structure of the N-terminal domain (residues 18–112) of α2-macroglobulin receptor-associated protein (RAP) has been determined by NMR spectroscopy. The structure consists of three helices composed of residues 23–34, 39–65, and 73–88. The three helices are arranged in an up-down-up antiparallel topology. The C-terminal 20 residues were shown not to be in a well defined conformation. A structural model for the binding of RAP to the family of low-density lipoprotein receptors is proposed. It defines a role in binding for both the unordered C terminus and the structural scaffold of the core structure. Pathogenic epitopes for the rat disease Heymann nephritis, an experimental model of human membranous glomerulonephritis, have been identified in RAP and in the large endocytic receptor gp330/megalin. Here we provide the three-dimensional structure of the pathogenic epitope in RAP. The amino acid residues known to form the epitope are in a helix–loop–helix conformation, and from the structure it is possible to rationalize the published results obtained from studies of fragments of the N-terminal domain.

Two Components of Actin-based Retrograde Flow in Sea Urchin Coelomocytes

Sea urchin coelomocytes represent an excellent experimental model system for studying retrograde flow. Their extreme flatness allows for excellent microscopic visualization. Their discoid shape provides a radially symmetric geometry, which simplifies analysis of the flow pattern. Finally, the nonmotile nature of the cells allows for the retrograde flow to be analyzed in the absence of cell translocation. In this study we have begun an analysis of the retrograde flow mechanism by characterizing its kinetic and structural properties. The supramolecular organization of actin and myosin II was investigated using light and electron microscopic methods. Light microscopic immunolocalization was performed with anti-actin and anti-sea urchin egg myosin II antibodies, whereas transmission electron microscopy was performed on platinum replicas of critical point-dried and rotary-shadowed cytoskeletons. Coelomocytes contain a dense cortical actin network, which feeds into an extensive array of radial bundles in the interior. These actin bundles terminate in a perinuclear region, which contains a ring of myosin II bipolar minifilaments. Retrograde flow was arrested either by interfering with actin polymerization or by inhibiting myosin II function, but the pathway by which the flow was blocked was different for the two kinds of inhibitory treatments. Inhibition of actin polymerization with cytochalasin D caused the actin cytoskeleton to separate from the cell margin and undergo a finite retrograde retraction. In contrast, inhibition of myosin II function either with the wide-spectrum protein kinase inhibitor staurosporine or the myosin light chain kinase–specific inhibitor KT5926 stopped flow in the cell center, whereas normal retrograde flow continued at the cell periphery. These differential results suggest that the mechanism of retrograde flow has two, spatially segregated components. We propose a “push–pull” mechanism in which actin polymerization drives flow at the cell periphery, whereas myosin II provides the tension on the actin cytoskeleton necessary for flow in the cell interior.

Mutagenesis associated with nitric oxide production in transgenic SJL mice

We recently reported development of an experimental model for the study of nitric oxide (NO·) toxicology in vivo. SJL mice were injected with superantigen-bearing RcsX (pre-B-cell lymphoma) cells, which migrated to the spleen and lymph nodes, where their rapid growth induced activation of macrophages to produce large amounts of NO· over a period of several weeks. In the experiments described here, we used this model to investigate mutagenesis in splenocytes exposed to NO· during RcsX cell growth. Transgenic mice were produced by crossbreeding animals of the pUR288 transgenic C57BL/6 and SJL strains. RcsX cells were injected into F1 mice and NO· production was confirmed by quantification of urinary nitrate, the ultimate metabolite of NO·. Mutant frequency in the lacZ gene of the pUR288 plasmid was determined in DNA isolated from spleen (target) and kidney (nontarget) tissues. A significant elevation in mutant frequency was found in the spleen, but not in the kidney, of tumor-bearing mice. Furthermore, increases in mutant frequency in the spleen as well as NO· production were abrogated by administration of N-methylarginine, a NO· inhibitor, to mice following injection of RcsX cells. These results indicate that NO· had mutagenic activity in RcsX tumor-bearing mice and thus support a possible role for its involvement in the carcinogenic process.

Identification of hepatitis B virus indigenous to chimpanzees

Hepatitis B viruses (HBV) and related viruses, classified in the Hepadnaviridae family, are found in a wide variety of mammals and birds. Although the chimpanzee has been the primary experimental model of HBV infection, this species has not been considered a natural host for the virus. Retrospective analysis of 13 predominantly wild-caught chimpanzees with chronic HBV infection identified a unique chimpanzee HBV strain in 11 animals. Nucleotide and derived amino acid analysis of the complete HBV genome and the gene coding for the hepatitis B surface antigen (S gene) identified sequence patterns that could be used to reliably identify chimpanzee HBV. This analysis indicated that chimpanzee HBV is distinct from known human HBV genotypes and is closely related to HBVs previously isolated from a chimpanzee, gibbons, gorillas, and orangutans.

Cholecystokinin-8 protects central cholinergic neurons against fimbria–fornix lesion through the up-regulation of nerve growth factor synthesis

In this study, we demonstrate that cholecystokinin-8 (CCK-8) induces an increase in both nerve growth factor (NGF) protein and NGF mRNA in mouse cortex and hippocampus when i.p. injected at physiological doses. By using fimbria–fornix-lesioned mice, we have also demonstrated that repeated CCK-8 i.p. injections result in recovery of lesion-induced NGF deficit in septum and restore the baseline NGF levels in hippocampus and cortex. Parallel to the effects on NGF, CCK-8 increases choline acetyltransferase (Chat) activity in forebrain when injected in unlesioned mice and counteract the septo-hippocampal Chat alterations in fimbria–fornix-lesioned mice. To assess the NGF involvement in the mechanism by which CCK-8 induces brain Chat, NGF antibody was administrated intracerebrally to saline- and CCK-8-injected mice. We observe that pretreatment with NGF antibody causes a marked reduction of NGF and Chat activity in septum and hippocampus of both saline- and CCK-8-injected mice. This evidence indicates that the CCK-8 effects on cholinergic cells are mediated through the synthesis and release of NGF. Taken together, our results suggest that peripheral administration of CCK-8 may represent a potential experimental model for investigating the effects of endogenous NGF up-regulation on diseases associated with altered brain cholinergic functions.

Emergency airway management by intensive care unit nurses with the intubating laryngeal mask airway and the laryngeal tube

When using the laryngeal tube and the intubating laryngeal mask airway (ILMA), the medium-size (maximum volume 1100 ml) versus adult (maximum volume 1500 ml) self-inflating bags resulted in significantly lower lung tidal volumes. No gastric inflation occurred when using both devices with either ventilation bag. The newly developed medium-size self-inflating bag may be an option to further reduce the risk of gastric inflation while maintaining sufficient lung ventilation. Both the ILMA and laryngeal tube proved to be valid alternatives for emergency airway management in the experimental model used.