19 resultados para local sequence alignment problem
Resumo:
The inbound logistic for feeding the workstation inside the factory represents a critical issue in the car manufacturing industry. Nowadays, this issue is even more critical than in the past since more types of car are being produced in the assembly lines. Consequently, as workstations have to install many types of components, they also need to have an inventory of different types of the component in a compact space. The replenishment is a critical issue since a lack of inventory could cause line stoppage or reworking. On the other hand, an excess of inventory could increase the holding cost or even block the replenishment paths. The decision of the replenishment routes cannot be made without taking into consideration the inventory needed by each station during the production time which will depend on the production sequence. This problem deals with medium-sized instances and it is solved using online solvers. The contribution of this paper is a MILP for the replenishment and inventory of the components in a car assembly line.
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This article focuses on the evaluation of a biometric technique based on the performance of an identifying gesture by holding a telephone with an embedded accelerometer in his/her hand. The acceleration signals obtained when users perform gestures are analyzed following a mathematical method based on global sequence alignment. In this article, eight different scores are proposed and evaluated in order to quantify the differences between gestures, obtaining an optimal EER result of 3.42% when analyzing a random set of 40 users of a database made up of 80 users with real attempts of falsification. Moreover, a temporal study of the technique is presented leeding to the need to update the template to adapt the manner in which users modify how they perform their identifying gesture over time. Six updating schemes have been assessed within a database of 22 users repeating their identifying gesture in 20 sessions over 4 months, concluding that the more often the template is updated the better and more stable performance the technique presents.
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This article proposes an innovative biometric technique based on the idea of authenticating a person on a mobile device by gesture recognition. To accomplish this aim, a user is prompted to be recognized by a gesture he/she performs moving his/her hand while holding a mobile device with an accelerometer embedded. As users are not able to repeat a gesture exactly in the air, an algorithm based on sequence alignment is developed to correct slight differences between repetitions of the same gesture. The robustness of this biometric technique has been studied within 2 different tests analyzing a database of 100 users with real falsifications. Equal Error Rates of 2.01 and 4.82% have been obtained in a zero-effort and an active impostor attack, respectively. A permanence evaluation is also presented from the analysis of the repetition of the gestures of 25 users in 10 sessions over a month. Furthermore, two different gesture databases have been developed: one made up of 100 genuine identifying 3-D hand gestures and 3 impostors trying to falsify each of them and another with 25 volunteers repeating their identifying 3- D hand gesture in 10 sessions over a month. These databases are the most extensive in published studies, to the best of our knowledge.
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Fastener holes in aeronautical structures are typical sources of fatigue cracks due to their induced local stress concentration. A very efficient solution to this problem is to establish compressive residual stresses around the fastener holes that retard the fatigue crack nucleation and its subsequent local propagation. Previous work done on the subject of the application of LSP treatment on thin, open-hole specimens [1] has proven that the LSP effect on fatigue life of treated specimens can be detrimental, if the process is not properly optimized. In fact, it was shown that the capability of the LSP to introduce compressive residual stresses around fastener holes in thin-walled structures representative of typical aircraft constructions was not superior to the performance of conventional techniques, such as cold-working.
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A proper allocation of resources targeted to solve hunger is essential to optimize the efficacy of actions and maximize results. This requires an adequate measurement and formulation of the problem as, paraphrasing Einstein, the formulation of a problem is essential to reach a solution. Different measurement methods have been designed to count, score, classify and compare hunger at local level and to allow comparisons between different places. However, the alternative methods produce significantly reach different results. These discrepancies make decisions on the targeting of resource allocations difficult. To assist decision makers, a new method taking into account the dimension of hunger and the coping capacities of countries, is proposed enabling to establish both geographical and sectoral priorities for the allocation of resources.
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A proper allocation of resources targeted to solve hunger is essential to optimize the efficacy of actions and maximize results. This requires an adequate measurement and formulation of the problem as, paraphrasing Einstein, the formulation of a problem is essential to reach a solution. Different measurement methods have been designed to count, score, classify and compare hunger at local level and to allow comparisons between different places. However, the alternative methods reach significantly different results. These discrepancies make decisions on the targeting of resource allocations difficult. To assist decision makers, a new method taking into account the dimension of hunger and the coping capacities of countries is proposed enabling to establish both geographical and sectoral priorities for the allocation of resources
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A local proper orthogonal decomposition (POD) plus Galerkin projection method was recently developed to accelerate time dependent numerical solvers of PDEs. This method is based on the combined use of a numerical code (NC) and a Galerkin sys- tem (GS) in a sequence of interspersed time intervals, INC and IGS, respectively. POD is performed on some sets of snapshots calculated by the numerical solver in the INC inter- vals. The governing equations are Galerkin projected onto the most energetic POD modes and the resulting GS is time integrated in the next IGS interval. The major computa- tional e®ort is associated with the snapshots calculation in the ¯rst INC interval, where the POD manifold needs to be completely constructed (it is only updated in subsequent INC intervals, which can thus be quite small). As the POD manifold depends only weakly on the particular values of the parameters of the problem, a suitable library can be con- structed adapting the snapshots calculated in other runs to drastically reduce the size of the ¯rst INC interval and thus the involved computational cost. The strategy is success- fully tested in (i) the one-dimensional complex Ginzburg-Landau equation, including the case in which it exhibits transient chaos, and (ii) the two-dimensional unsteady lid-driven cavity problem
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Se desarrollan varias técnicas basadas en descomposición ortogonal propia (DOP) local y proyección de tipo Galerkin para acelerar la integración numérica de problemas de evolución, de tipo parabólico, no lineales. Las ideas y métodos que se presentan conllevan un nuevo enfoque para la modelización de tipo DOP, que combina intervalos temporales cortos en que se usa un esquema numérico estándard con otros intervalos temporales en que se utilizan los sistemas de tipo Galerkin que resultan de proyectar las ecuaciones de evolución sobre la variedad lineal generada por los modos DOP, obtenidos a partir de instantáneas calculadas en los intervalos donde actúa el código numérico. La variedad DOP se construye completamente en el primer intervalo, pero solamente se actualiza en los demás intervalos según las dinámicas de la solución, aumentando de este modo la eficiencia del modelo de orden reducido resultante. Además, se aprovechan algunas propiedades asociadas a la dependencia débil de los modos DOP tanto en la variable temporal como en los posibles parámetros de que pueda depender el problema. De esta forma, se aumentan la flexibilidad y la eficiencia computacional del proceso. La aplicación de los métodos resultantes es muy prometedora, tanto en la simulación de transitorios en flujos laminares como en la construcción de diagramas de bifurcación en sistemas dependientes de parámetros. Las ideas y los algoritmos desarrollados en la tesis se ilustran en dos problemas test, la ecuación unidimensional compleja de Ginzburg-Landau y el problema bidimensional no estacionario de la cavidad. Abstract Various ideas and methods involving local proper orthogonal decomposition (POD) and Galerkin projection are presented aiming at accelerating the numerical integration of nonlinear time dependent parabolic problems. The proposed methods come from a new approach to the POD-based model reduction procedures, which combines short runs with a given numerical solver and a reduced order model constructed by expanding the solution of the problem into appropriate POD modes, which span a POD manifold, and Galerkin projecting some evolution equations onto that linear manifold. The POD manifold is completely constructed from the outset, but only updated as time proceeds according to the dynamics, which yields an adaptive and flexible procedure. In addition, some properties concerning the weak dependence of the POD modes on time and possible parameters in the problem are exploited in order to increase the flexibility and efficiency of the low dimensional model computation. Application of the developed techniques to the approximation of transients in laminar fluid flows and the simulation of attractors in bifurcation problems shows very promising results. The test problems considered to illustrate the various ideas and check the performance of the algorithms are the onedimensional complex Ginzburg-Landau equation and the two-dimensional unsteady liddriven cavity problem.
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Many macroscopic properties: hardness, corrosion, catalytic activity, etc. are directly related to the surface structure, that is, to the position and chemical identity of the outermost atoms of the material. Current experimental techniques for its determination produce a “signature” from which the structure must be inferred by solving an inverse problem: a solution is proposed, its corresponding signature computed and then compared to the experiment. This is a challenging optimization problem where the search space and the number of local minima grows exponentially with the number of atoms, hence its solution cannot be achieved for arbitrarily large structures. Nowadays, it is solved by using a mixture of human knowledge and local search techniques: an expert proposes a solution that is refined using a local minimizer. If the outcome does not fit the experiment, a new solution must be proposed again. Solving a small surface can take from days to weeks of this trial and error method. Here we describe our ongoing work in its solution. We use an hybrid algorithm that mixes evolutionary techniques with trusted region methods and reuses knowledge gained during the execution to avoid repeated search of structures. Its parallelization produces good results even when not requiring the gathering of the full population, hence it can be used in loosely coupled environments such as grids. With this algorithm, the solution of test cases that previously took weeks of expert time can be automatically solved in a day or two of uniprocessor time.
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This paper studies the problem of determining the position of beacon nodes in Local Positioning Systems (LPSs), for which there are no inter-beacon distance measurements available and neither the mobile node nor any of the stationary nodes have positioning or odometry information. The common solution is implemented using a mobile node capable of measuring its distance to the stationary beacon nodes within a sensing radius. Many authors have implemented heuristic methods based on optimization algorithms to solve the problem. However, such methods require a good initial estimation of the node positions in order to find the correct solution. In this paper we present a new method to calculate the inter-beacon distances, and hence the beacons positions, based in the linearization of the trilateration equations into a closed-form solution which does not require any approximate initial estimation. The simulations and field evaluations show a good estimation of the beacon node positions.
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In the context of aerial imagery, one of the first steps toward a coherent processing of the information contained in multiple images is geo-registration, which consists in assigning geographic 3D coordinates to the pixels of the image. This enables accurate alignment and geo-positioning of multiple images, detection of moving objects and fusion of data acquired from multiple sensors. To solve this problem there are different approaches that require, in addition to a precise characterization of the camera sensor, high resolution referenced images or terrain elevation models, which are usually not publicly available or out of date. Building upon the idea of developing technology that does not need a reference terrain elevation model, we propose a geo-registration technique that applies variational methods to obtain a dense and coherent surface elevation model that is used to replace the reference model. The surface elevation model is built by interpolation of scattered 3D points, which are obtained in a two-step process following a classical stereo pipeline: first, coherent disparity maps between image pairs of a video sequence are estimated and then image point correspondences are back-projected. The proposed variational method enforces continuity of the disparity map not only along epipolar lines (as done by previous geo-registration techniques) but also across them, in the full 2D image domain. In the experiments, aerial images from synthetic video sequences have been used to validate the proposed technique.
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A generalized Lévêque solution is presented for the conjugate fluid–fluid problem that arises in the thermal entrance region of laminar counterflow heat exchangers. The analysis, carried out for constant property fluids, assumes that the Prandtl and Peclet numbers are both large compared to unity, and neglects axial conduction both in the fluids and in the plate, assumed to be thermally thin. Under these conditions, the thermal entrance region admits an asymptotic self-similar description where the temperature varies as a power ϳ of the axial distance, with the particularity that the self-similarity exponent must be determined as an eigenvalue by solving a transcendental equation arising from the requirement of continuity of heat fluxes at the heat conducting wall. Specifically, the analysis reveals that j depends only on the lumped parameter ƙ = (A2/A1)1/3 (α1/α2)1/3(k2/k1), defined in terms of the ratios of the wall velocity gradients, A, thermal diffusivities, α i, and thermal conductivities,k i, of the fluids entering, 1, and exiting, 2, the heat exchanger. Moreover, it is shown that for large (small) values of K solution reduces to the classical first (second) Lévêque solution. Closed-form analytical expressions for the asymptotic temperature distributions and local heat-transfer rate in the thermal entrance region are given and compared with numerical results in the counterflow parallel-plate configuration, showing very good agreement in all cases.
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El aprendizaje basado en problemas se lleva aplicando con éxito durante las últimas tres décadas en un amplio rango de entornos de aprendizaje. Este enfoque educacional consiste en proponer problemas a los estudiantes de forma que puedan aprender sobre un dominio particular mediante el desarrollo de soluciones a dichos problemas. Si esto se aplica al modelado de conocimiento, y en particular al basado en Razonamiento Cualitativo, las soluciones a los problemas pasan a ser modelos que representan el compotamiento del sistema dinámico propuesto. Por lo tanto, la tarea del estudiante en este caso es acercar su modelo inicial (su primer intento de representar el sistema) a los modelos objetivo que proporcionan soluciones al problema, a la vez que adquieren conocimiento sobre el dominio durante el proceso. En esta tesis proponemos KaiSem, un método que usa tecnologías y recursos semánticos para guiar a los estudiantes durante el proceso de modelado, ayudándoles a adquirir tanto conocimiento como sea posible sin la directa supervisión de un profesor. Dado que tanto estudiantes como profesores crean sus modelos de forma independiente, estos tendrán diferentes terminologías y estructuras, dando lugar a un conjunto de modelos altamente heterogéneo. Para lidiar con tal heterogeneidad, proporcionamos una técnica de anclaje semántico para determinar, de forma automática, enlaces entre la terminología libre usada por los estudiantes y algunos vocabularios disponibles en la Web de Datos, facilitando con ello la interoperabilidad y posterior alineación de modelos. Por último, proporcionamos una técnica de feedback semántico para comparar los modelos ya alineados y generar feedback basado en las posibles discrepancias entre ellos. Este feedback es comunicado en forma de sugerencias individualizadas que el estudiante puede utilizar para acercar su modelo a los modelos objetivos en cuanto a su terminología y estructura se refiere. ABSTRACT Problem-based learning has been successfully applied over the last three decades to a diverse range of learning environments. This educational approach consists of posing problems to learners, so they can learn about a particular domain by developing solutions to them. When applied to conceptual modeling, and particularly to Qualitative Reasoning, the solutions to problems are models that represent the behavior of a dynamic system. Therefore, the learner's task is to move from their initial model, as their first attempt to represent the system, to the target models that provide solutions to that problem while acquiring domain knowledge in the process. In this thesis we propose KaiSem, a method for using semantic technologies and resources to scaffold the modeling process, helping the learners to acquire as much domain knowledge as possible without direct supervision from the teacher. Since learners and experts create their models independently, these will have different terminologies and structure, giving rise to a pool of models highly heterogeneous. To deal with such heterogeneity, we provide a semantic grounding technique to automatically determine links between the unrestricted terminology used by learners and some online vocabularies of the Web of Data, thus facilitating the interoperability and later alignment of the models. Lastly, we provide a semantic-based feedback technique to compare the aligned models and generate feedback based on the possible discrepancies. This feedback is communicated in the form of individualized suggestions, which can be used by the learner to bring their model closer in terminology and structure to the target models.
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Among the different interchange design aspects, integrated land use and infrastructure planning is maybe one of the most problematic fields in practice, given that a joint transport and urban planning spills over the regular scope of action of interchange developers, whereas it involves the cooperation and agreement of various authorities. Not only this, but the very issue of land use-transport integration seems to be a long-standing mantra in planning and transport research, lacking scientific evidence. This paper is an output of an ongoing European research project called ?NODES - New tOols for Design and OpEration of Urban Transport InterchangeS?. Its aim is to start re-focusing the academic-scientific evidence on the question and to foresee a specific and practical framework to approach the problem. The underlying hypothesis is that the interchange could be a catalyst of life and security in the city.
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Cinchona officinalis (Rubiaceae), especie endémica del Valle de Loja, ubicado en la región sur del Ecuador, es un recurso forestal de importancia medicinal y ecológica, además la especie ha sido catalogada como planta nacional y es un ícono de la región sur por su aporte a la farmacopea mundial. Esta especie, entre los siglos XVII-XIX sufrió una gran presión en sus poblaciones debido a la extracción masiva de la corteza para la cura del paludismo. Aunque la actividad extractiva generó grandes ingresos a la Corona Española y a la región Sur del Ecuador, ésta fue poco o nada sustentable ecológicamente, provocando la desaparición de la especie en muchos sitios de la provincia, pues, en su momento, no se consideraron alternativas de recuperación de las poblaciones naturales. Actualmente la extracción y consumo de la corteza en la zona de origen es baja o nula, sin embargo esta zona enfrenta nuevas amenazas. La deforestación a causa de proyectos de desarrollo en infraestructuras, la práctica de actividades agrícolas y de ganadería, y los efectos del cambio climático han ocasionado, en estos últimos años, la fragmentación de los ecosistemas. La mayoría de los bosques del sur del Ecuador se han convertido en parches aislados (los bosques en los que se distribuye C. officinalis no son la excepción) siendo esta la principal causa para que la especie se encuentre en estado de amenaza. Los individuos de la especie tienen una alta capacidad de rebrote y producen semillas durante todo el año; sin embargo la capacidad germinativa y la tasa de sobrevivencia son bajas, además de estas dificultades la especie requiere de la asociación con otras especies vegetales para su desarrollo, lo cual ha limitado su distribución en pequeños parches aislados. Con esta problemática, la recuperación natural de las poblaciones es una necesidad evidente. Varios trabajos y esfuerzos previos se han realizado a nivel local: i. Identificación de la distribución actual y potencial; ii. Determinación de la fenología y fructificación iii. Programas de educación ambiental, iv. Análisis moleculares para determinar la diversidad genética. v. Ensayos de propagación vegetativa; y otras acciones de tipo cultural. No obstante, el estado de conservación y manejo de las poblaciones naturales no ha mejorado significativamente, siendo necesaria la aplicación de estrategias integradas de conservación in situ y ex situ, que permitan la recuperación y permanencia de las poblaciones naturales a largo plazo. El presente trabajo tiene como fin dar alternativas para el cultivo de tejidos in vitro de Cinchona officinalis centrados en la propagación masiva a partir de semillas, análisis de la fidelidad genética y alternativas de conservación de tejidos. Los objetivos específicos que se plantean son: i. Analizar el proceso de germinación y proliferación in vitro. ii. Evaluar la estabilidad genética en explantes cultivados in vitro, mediante marcadores ISSR. iii. Establecer protocolos de conservación in vitro mediante limitación del crecimiento y criopreservación de segmentos nodales y yemas. Los resultados más significativos de esta investigación fueron: i. El desarrollo de protocolos eficientes para mejorar los porcentajes de germinación y la proliferación de brotes en explantos cultivados in vitro. Para evaluar el efecto de los fenoles sobre la germinación, se determinó el contenido total de fenoles y el porcentaje de germinación en semillas de C. officinalis comparados con una especie de control, C. pubescens. Para inducir a proliferación, se utilizaron segmentos nodales de plántulas germinadas in vitro en medio Gamborg (1968) suplementado con diferentes combinaciones de reguladores de crecimiento (auxinas y citoquininas). Los resultados obtenidos sugieren que el contenido de compuestos fenólicos es alto en las semillas de C. officinalis en comparación con las semillas de C. pubescens. Estos fenoles pueden eliminarse con peróxido de hidrógeno o con lavados de agua para estimular la germinación. La formación de nuevos brotes y callos en la mayoría de las combinaciones de reguladores de crecimiento se observó en un período de 45 días. El mayor porcentaje de proliferación de brotes, formación de callos y presencia de brotes adventicios se obtuvo en medio Gamborg (B5) suplementado con 5.0 mg/l 6-bencil-aminopurina y 3.0 mg/l de ácido indol-3-butírico. ii. La evaluación de la fidelidad genética de los explantes obtenidos con distintas combinaciones de reguladores de crecimiento vegetal y diversos subcultivos. Se realizó el seguimiento a los explantes obtenidos de la fase anterior, determinando el índice de multiplicación y analizando la fidelidad genética de los tejidos obtenidos por las dos vías regenerativas: brotación directa y regeneración de brotes a partir de callos. Este análisis se realizó por amplificación mediante PCR de las secuencias ubicadas entre microsatélites-ISSR (Inter simple sequence repeat). El medio Gamborg (B5) con 3.0 mg/l de AIB y 5.0 mg/l de BAP usado como medio de inducción en la primera etapa de cultivo generó el mayor índice de proliferación (11.5). Un total de 13 marcadores ISSR fueron analizados, 6 de éstos fueron polimórficos. El mayor porcentaje de variación somaclonal fue inducido en presencia de 1.0 mg/l 2,4-D combinado con 0.2 mg/l Kin con un 1.8% en el segundo sub-cultivo de regeneración, la cual incrementó a 3.6% en el tercer sub-cultivo. Todas las combinaciones con presencia de 2,4-D produjeron la formación de callos y presentaron variación genética. Por su parte la fidelidad genética se mantuvo en los sistemas de propagación directa a través de la formación de brotes a partir de meristemos preformados. iii. El establecimiento de protocolos de conservación in vitro y crioconservación de segmentos nodales y yemas. Para la conservación limitando el crecimiento, se cultivaron segmentos nodales en los medios MS y B5 en tres concentraciones de sus componentes (25, 50 y 100%); y en medio B5 más agentes osmóticos como el manitol, sorbitol y sacarosa en diferentes concentraciones (2, 4 y 8%); los cultivos se mantuvieron por 12 meses sin subcultivos. Para el establecimiento de protocolos para la crioconservación (paralización del metabolismo) se usaron yemas axilares y apicales a las cuales se les aplicaron los métodos de encapsulación-deshidratación y vitrificación. La efectividad de los protocolos usados se determinó en función de la sobrevivencia, reducción del crecimiento y regeneración. Los resultados obtenidos en este apartado reflejan que un crecimiento limitado puede mantener tejidos durante 12 meses de almacenamiento, usando medio B5 más manitol entre 2 y 8%. En los protocolos de crioconservación, se obtuvo el mayor porcentaje de recuperación tras la congelación en NL en el tratamiento control seguido por el método crioprotector de encapsulación-deshidratación. Este trabajo brinda alternativas para la propagación de C. officinalis bajo condiciones in vitro, partiendo de material vegetal con alta diversidad genética. El material propagado puede ser fuente de germoplasma para la recuperación y reforzamiento de las poblaciones naturales así como una alternativa de producción para las comunidades locales debido a la demanda actual de corteza de la zona de origen para la elaboración de agua tónica. ABSTRACT Cinchona officinalis (Rubiaceae) is endemic to the Loja Valley, located in the southern area of Ecuador. The importance of this plant as medical and ecological resource is so great that it has been designated as the national flower and is an icon of the southern region for its contribution to the world pharmacopoeia. Between XVII-XIX centuries its population suffered great reduction due to massive harvesting of the bark to cure malaria. Although extraction activity generated large revenues to the Spanish Crown and the southern region of Ecuador, this was not ecologically sustainable, causing the disappearance of the species in many areas of the province, because during that time alternatives to prevent extinction and recover natural populations were not taken in account. Currently the extraction and consumption of bark in the area of origin is almost absent, but this species faces new threats. Deforestation due to infrastructure development, the practice of farming and ranching, and the effects of climate change had led to the fragmentation of ecosystems during the recent years. Most of the forests of southern Ecuador have become isolated patches, including those where C. officinalis is diffused. The lack of suitable habitat is today the main threat for the species. The species has a high capacity for regeneration and produces seeds throughout the year, but the germination rate is low and the growth is slow. In addition, the species requires the association with other plant species to develop. All these factors had limited its distribution to small isolated patches. The natural recovery of populations is essential to face this problem. Several studies and previous efforts had been made at local level: i. Identification of current and potential distribution; ii. Phenology determination. iii. Environmental education programs, iv. Molecular analisis to determine the genetic diversity. v. Testing of vegetative propagation; and other actions of cultural nature. Despite these efforts, the state of conservation and management of natural populations has not improved significantly. Implementation of integrated in situ and ex situ conservation strategies for the recovery and permanence of long-term natural populations is still needed. This work aims to provide alternatives for in vitro culture of tissue of Cinchona officinalis focused on mass propagation from seeds, genetic fidelity analysis and tissue conservation alternatives. The specific aims are: i. Analyze the process of germination and proliferation in vitro. ii. To evaluate the genetic stability of the explants cultured in vitro by ISSR markers. iii. Establish protocols for in vitro conservation by limiting growth and cryopreservation of nodal segments and buds. The most significant results of this research were: i. The development of efficient protocols to improve germination rates and proliferation of buds in explants cultured in vitro. To study the effect of phenols on germination, the total phenolic content and percentage germination was measured in C. officinalis and in a control species, C. pubescens, for comparison. The content of phenolic compounds in C. officinalis seeds is higher than in C. pubescens. These phenols can be removed with hydrogen peroxide or water washes to stimulate germination. To analyze the regeneration, we used nodal explants from seedlings germinated in vitro on Gamborg medium (1968) supplemented with different combinations of growth regulators (auxins and cytokinins) to induce proliferation. The formation of new shoots and calluses was observed within a period of 45 days in most combinations of growth regulators. The highest percentage of shoot proliferation, callus formation and adventitious buds were obtained in B5 medium supplemented with 5.0 mg/l 6-benzyl-aminopurine and 3.0 mg/l indole-3-butyric acid. ii. Evaluating genetic fidelity explants obtained with various combinations of plant growth regulators and different subcultures. The genetic fidelity was analyzed in tissues obtained by the two regenerative pathways: direct sprouting and shoot regeneration from callus. This analysis was performed by PCR amplification of the sequences located between microsatellite-ISSR (Inter Simple Sequence Repeat). Among a total of 13 ISSR markers analyzed, 6 were polymorphic. The highest percentage of somaclonal variation was induced in the presence of 1.0 mg/l 2,4-D combined with 0.2 mg/l Kin with 1.8% in the second round of regeneration, and increased to 3.6% in the third round. The presence of 2,4-D induced genetic variation in all the combinations of growth regulators. Meanwhile genetic fidelity remained systems propagation through direct shoot formation from meristems preformed. iii. Establishing conservation protocols in vitro and cryoconservation of nodal segments and buds. For medium-term conservation (limited growth) nodal segments were cultured in MS and B5 media at three concentrations (25, 50 and 100%); we tested B5 medium with different concentrations of osmotic agents such as mannitol, sorbitol and sucrose (2, 4 and 8%); cultures were maintained for 12 months with regular subculturing. To establish protocols for cryoconservation (cessation of metabolism) different methods of encapsulation-dehydration and vitrification were applied to axillary and apical buds. The effectiveness of the used protocols is determined based on the survival, growth and regeneration success. The results show that these tissues can be maintained in storage for 12 months, using B5 medium plus mannitol between 2 and 8%. The cryoconservation protocol with highest percentage of recovery was obtained by contral treatment, followed by freezing in NL with encapsulation-dehydration method. This work provides alternatives for the propagation in vitro of C. officinalis, starting from plant material with high genetic diversity. The obtained material represents a source of germplasm to support the recovery and strengthening of natural populations as well as a creation of alternative sources for local communities due to the current demand of bark for the preparation of tonic water.