Methods and Tools for Objective Assessment of Psychomotor Skills in Laparoscopic Surgery

Training and assessment paradigms for laparoscopic surgical skills are evolving from traditional mentor–trainee tutorship towards structured, more objective and safer programs. Accreditation of surgeons requires reaching a consensus on metrics and tasks used to assess surgeons’ psychomotor skills. Ongoing development of tracking systems and software solutions has allowed for the expansion of novel training and assessment means in laparoscopy. The current challenge is to adapt and include these systems within training programs, and to exploit their possibilities for evaluation purposes. This paper describes the state of the art in research on measuring and assessing psychomotor laparoscopic skills. It gives an overview on tracking systems as well as on metrics and advanced statistical and machine learning techniques employed for evaluation purposes. The later ones have a potential to be used as an aid in deciding on the surgical competence level, which is an important aspect when accreditation of the surgeons in particular, and patient safety in general, are considered. The prospective of these methods and tools make them complementary means for surgical assessment of motor skills, especially in the early stages of training. Successful examples such as the Fundamentals of Laparoscopic Surgery should help drive a paradigm change to structured curricula based on objective parameters. These may improve the accreditation of new surgeons, as well as optimize their already overloaded training schedules.

Conceptual framework for the design, implementation and validation of psychomotor skills’ assessment systems in minimally invasive surgery

Las técnicas de cirugía de mínima invasión (CMI) se están consolidando hoy en día como alternativa a la cirugía tradicional, debido a sus numerosos beneficios para los pacientes. Este cambio de paradigma implica que los cirujanos deben aprender una serie de habilidades distintas de aquellas requeridas en cirugía abierta. El entrenamiento y evaluación de estas habilidades se ha convertido en una de las mayores preocupaciones en los programas de formación de cirujanos, debido en gran parte a la presión de una sociedad que exige cirujanos bien preparados y una reducción en el número de errores médicos. Por tanto, se está prestando especial atención a la definición de nuevos programas que permitan el entrenamiento y la evaluación de las habilidades psicomotoras en entornos seguros antes de que los nuevos cirujanos puedan operar sobre pacientes reales. Para tal fin, hospitales y centros de formación están gradualmente incorporando instalaciones de entrenamiento donde los residentes puedan practicar y aprender sin riesgos. Es cada vez más común que estos laboratorios dispongan de simuladores virtuales o simuladores físicos capaces de registrar los movimientos del instrumental de cada residente. Estos simuladores ofrecen una gran variedad de tareas de entrenamiento y evaluación, así como la posibilidad de obtener información objetiva de los ejercicios. Los diferentes estudios de validación llevados a cabo dan muestra de su utilidad; pese a todo, los niveles de evidencia presentados son en muchas ocasiones insuficientes. Lo que es más importante, no existe un consenso claro a la hora de definir qué métricas son más útiles para caracterizar la pericia quirúrgica. El objetivo de esta tesis doctoral es diseñar y validar un marco de trabajo conceptual para la definición y validación de entornos para la evaluación de habilidades en CMI, en base a un modelo en tres fases: pedagógica (tareas y métricas a emplear), tecnológica (tecnologías de adquisición de métricas) y analítica (interpretación de la competencia en base a las métricas). Para tal fin, se describe la implementación práctica de un entorno basado en (1) un sistema de seguimiento de instrumental fundamentado en el análisis del vídeo laparoscópico; y (2) la determinación de la pericia en base a métricas de movimiento del instrumental. Para la fase pedagógica se diseñó e implementó un conjunto de tareas para la evaluación de habilidades psicomotoras básicas, así como una serie de métricas de movimiento. La validación de construcción llevada a cabo sobre ellas mostró buenos resultados para tiempo, camino recorrido, profundidad, velocidad media, aceleración media, economía de área y economía de volumen. Adicionalmente, los resultados obtenidos en la validación de apariencia fueron en general positivos en todos los grupos considerados (noveles, residentes, expertos). Para la fase tecnológica, se introdujo el EVA Tracking System, una solución para el seguimiento del instrumental quirúrgico basado en el análisis del vídeo endoscópico. La precisión del sistema se evaluó a 16,33ppRMS para el seguimiento 2D de la herramienta en la imagen; y a 13mmRMS para el seguimiento espacial de la misma. La validación de construcción con una de las tareas de evaluación mostró buenos resultados para tiempo, camino recorrido, profundidad, velocidad media, aceleración media, economía de área y economía de volumen. La validación concurrente con el TrEndo® Tracking System por su parte presentó valores altos de correlación para 8 de las 9 métricas analizadas. Finalmente, para la fase analítica se comparó el comportamiento de tres clasificadores supervisados a la hora de determinar automáticamente la pericia quirúrgica en base a la información de movimiento del instrumental, basados en aproximaciones lineales (análisis lineal discriminante, LDA), no lineales (máquinas de soporte vectorial, SVM) y difusas (sistemas adaptativos de inferencia neurodifusa, ANFIS). Los resultados muestran que en media SVM presenta un comportamiento ligeramente superior: 78,2% frente a los 71% y 71,7% obtenidos por ANFIS y LDA respectivamente. Sin embargo las diferencias estadísticas medidas entre los tres no fueron demostradas significativas. En general, esta tesis doctoral corrobora las hipótesis de investigación postuladas relativas a la definición de sistemas de evaluación de habilidades para cirugía de mínima invasión, a la utilidad del análisis de vídeo como fuente de información y a la importancia de la información de movimiento de instrumental a la hora de caracterizar la pericia quirúrgica. Basándose en estos cimientos, se han de abrir nuevos campos de investigación que contribuyan a la definición de programas de formación estructurados y objetivos, que puedan garantizar la acreditación de cirujanos sobradamente preparados y promocionen la seguridad del paciente en el quirófano. Abstract Minimally invasive surgery (MIS) techniques have become a standard in many surgical sub-specialties, due to their many benefits for patients. However, this shift in paradigm implies that surgeons must acquire a complete different set of skills than those normally attributed to open surgery. Training and assessment of these skills has become a major concern in surgical learning programmes, especially considering the social demand for better-prepared professionals and for the decrease of medical errors. Therefore, much effort is being put in the definition of structured MIS learning programmes, where practice with real patients in the operating room (OR) can be delayed until the resident can attest for a minimum level of psychomotor competence. To this end, skills’ laboratory settings are being introduced in hospitals and training centres where residents may practice and be assessed on their psychomotor skills. Technological advances in the field of tracking technologies and virtual reality (VR) have enabled the creation of new learning systems such as VR simulators or enhanced box trainers. These systems offer a wide range of tasks, as well as the capability of registering objective data on the trainees’ performance. Validation studies give proof of their usefulness; however, levels of evidence reported are in many cases low. More importantly, there is still no clear consensus on topics such as the optimal metrics that must be used to assess competence, the validity of VR simulation, the portability of tracking technologies into real surgeries (for advanced assessment) or the degree to which the skills measured and obtained in laboratory environments transfer to the OR. The purpose of this PhD is to design and validate a conceptual framework for the definition and validation of MIS assessment environments based on a three-pillared model defining three main stages: pedagogical (tasks and metrics to employ), technological (metric acquisition technologies) and analytical (interpretation of competence based on metrics). To this end, a practical implementation of the framework is presented, focused on (1) a video-based tracking system and (2) the determination of surgical competence based on the laparoscopic instruments’ motionrelated data. The pedagogical stage’s results led to the design and implementation of a set of basic tasks for MIS psychomotor skills’ assessment, as well as the definition of motion analysis parameters (MAPs) to measure performance on said tasks. Validation yielded good construct results for parameters such as time, path length, depth, average speed, average acceleration, economy of area and economy of volume. Additionally, face validation results showed positive acceptance on behalf of the experts, residents and novices. For the technological stage the EVA Tracking System is introduced. EVA provides a solution for tracking laparoscopic instruments from the analysis of the monoscopic video image. Accuracy tests for the system are presented, which yielded an average RMSE of 16.33pp for 2D tracking of the instrument on the image and of 13mm for 3D spatial tracking. A validation experiment was conducted using one of the tasks and the most relevant MAPs. Construct validation showed significant differences for time, path length, depth, average speed, average acceleration, economy of area and economy of volume; especially between novices and residents/experts. More importantly, concurrent validation with the TrEndo® Tracking System presented high correlation values (>0.7) for 8 of the 9 MAPs proposed. Finally, the analytical stage allowed comparing the performance of three different supervised classification strategies in the determination of surgical competence based on motion-related information. The three classifiers were based on linear (linear discriminant analysis, LDA), non-linear (support vector machines, SVM) and fuzzy (adaptive neuro fuzzy inference systems, ANFIS) approaches. Results for SVM show slightly better performance than the other two classifiers: on average, accuracy for LDA, SVM and ANFIS was of 71.7%, 78.2% and 71% respectively. However, when confronted, no statistical significance was found between any of the three. Overall, this PhD corroborates the investigated research hypotheses regarding the definition of MIS assessment systems, the use of endoscopic video analysis as the main source of information and the relevance of motion analysis in the determination of surgical competence. New research fields in the training and assessment of MIS surgeons can be proposed based on these foundations, in order to contribute to the definition of structured and objective learning programmes that guarantee the accreditation of well-prepared professionals and the promotion of patient safety in the OR.

EVA: Laparoscopic instrument tracking based on endoscopic video analysis for psychomotor skills assessment

INTRODUCTION: The EVA (Endoscopic Video Analysis) tracking system a new tracking system for extracting motions of laparoscopic instruments based on non-obtrusive video tracking was developed. The feasibility of using EVA in laparoscopic settings has been tested in a box trainer setup. METHODS: EVA makes use of an algorithm that employs information of the laparoscopic instrument's shaft edges in the image, the instrument's insertion point, and the camera's optical centre to track the 3D position of the instrument tip. A validation study of EVA comprised a comparison of the measurements achieved with EVA and the TrEndo tracking system. To this end, 42 participants (16 novices, 22 residents, and 4 experts) were asked to perform a peg transfer task in a box trainer. Ten motion-based metrics were used to assess their performance. RESULTS: Construct validation of the EVA has been obtained for seven motion-based metrics. Concurrent validation revealed that there is a strong correlation between the results obtained by EVA and the TrEndo for metrics such as path length (p=0,97), average speed (p=0,94) or economy of volume (p=0,85), proving the viability of EVA. CONCLUSIONS: EVA has been successfully used in the training setup showing potential of endoscopic video analysis to assess laparoscopic psychomotor skills. The results encourage further implementation of video tracking in training setups and in image guided surgery.

Learning curves of basic laparoscopic psychomotor skills in SINERGIA VR simulator

Purpose: Surgical simulators are currently essential within any laparoscopic training program because they provide a low-stakes, reproducible and reliable environment to acquire basic skills. The purpose of this study is to determine the training learning curve based on different metrics corresponding to five tasks included in SINERGIA laparoscopic virtual reality simulator. Methods: Thirty medical students without surgical experience participated in the study. Five tasks of SINERGIA were included: Coordination, Navigation, Navigation and touch, Accurate grasping and Coordinated pulling. Each participant was trained in SINERGIA. This training consisted of eight sessions (R1–R8) of the five mentioned tasks and was carried out in two consecutive days with four sessions per day. A statistical analysis was made, and the results of R1, R4 and R8 were pair-wise compared with Wilcoxon signed-rank test. Significance is considered at P value <0.005. Results: In total, 84.38% of the metrics provided by SINERGIA and included in this study show significant differences when comparing R1 and R8. Metrics are mostly improved in the first session of training (75.00% when R1 and R4 are compared vs. 37.50% when R4 and R8 are compared). In tasks Coordination and Navigation and touch, all metrics are improved. On the other hand, Navigation just improves 60% of the analyzed metrics. Most learning curves show an improvement with better results in the fulfillment of the different tasks. Conclusions: Learning curves of metrics that assess the basic psychomotor laparoscopic skills acquired in SINERGIA virtual reality simulator show a faster learning rate during the first part of the training. Nevertheless, eight repetitions of the tasks are not enough to acquire all psychomotor skills that can be trained in SINERGIA. Therefore, and based on these results together with previous works, SINERGIA could be used as training tool with a properly designed training program.

Adaptive neuro-fuzzy inference systems (ANFIS) for supervised classification of competence in MIS based on psychomotor skills

INTRODUCTION: Objective assessment of motor skills has become an important challenge in minimally invasive surgery (MIS) training.Currently, there is no gold standard defining and determining the residents' surgical competence.To aid in the decision process, we analyze the validity of a supervised classifier to determine the degree of MIS competence based on assessment of psychomotor skills METHODOLOGY: The ANFIS is trained to classify performance in a box trainer peg transfer task performed by two groups (expert/non expert). There were 42 participants included in the study: the non-expert group consisted of 16 medical students and 8 residents (< 10 MIS procedures performed), whereas the expert group consisted of 14 residents (> 10 MIS procedures performed) and 4 experienced surgeons. Instrument movements were captured by means of the Endoscopic Video Analysis (EVA) tracking system. Nine motion analysis parameters (MAPs) were analyzed, including time, path length, depth, average speed, average acceleration, economy of area, economy of volume, idle time and motion smoothness. Data reduction was performed by means of principal component analysis, and then used to train the ANFIS net. Performance was measured by leave one out cross validation. RESULTS: The ANFIS presented an accuracy of 80.95%, where 13 experts and 21 non-experts were correctly classified. Total root mean square error was 0.88, while the area under the classifiers' ROC curve (AUC) was measured at 0.81. DISCUSSION: We have shown the usefulness of ANFIS for classification of MIS competence in a simple box trainer exercise. The main advantage of using ANFIS resides in its continuous output, which allows fine discrimination of surgical competence. There are, however, challenges that must be taken into account when considering use of ANFIS (e.g. training time, architecture modeling). Despite this, we have shown discriminative power of ANFIS for a low-difficulty box trainer task, regardless of the individual significances between MAPs. Future studies are required to confirm the findings, inclusion of new tasks, conditions and sample population.

EVA: Endoscopic Video Analysis of the surgical scene for the assessment of MIS psychomotor skills

The present work covers the first validation efforts of the EVA Tracking System for the assessment of minimally invasive surgery (MIS) psychomotor skills. Instrument movements were recorded for 42 surgeons (4 expert, 22 residents, 16 novice medical students) and analyzed for a box trainer peg transfer task. Construct validation was established for 7/9 motion analysis parameters (MAPs). Concurrent validation was determined for 8/9 MAPs against the TrEndo Tracking System. Finally, automatic determination of surgical proficiency based on the MAPs was sought by 3 different approaches to supervised classification (LDA, SVM, ANFIS), with accuracy results of 61.9%, 83.3% and 80.9% respectively. Results not only reflect on the validation of EVA for skills? assessment, but also on the relevance of motion analysis of instruments in the determination of surgical competence.

Supervised classification of minimally invasive surgery psychomotor competence based on instruments motion analysis

Background Objective assessment of psychomotor skills has become an important challenge in the training of minimally invasive surgical (MIS) techniques. Currently, no gold standard defining surgical competence exists for classifying residents according to their surgical skills. Supervised classification has been proposed as a means for objectively establishing competence thresholds in psychomotor skills evaluation. This report presents a study comparing three classification methods for establishing their validity in a set of tasks for basic skills’ assessment. Methods Linear discriminant analysis (LDA), support vector machines (SVM), and adaptive neuro-fuzzy inference systems (ANFIS) were used. A total of 42 participants, divided into an experienced group (4 expert surgeons and 14 residents with >10 laparoscopic surgeries performed) and a nonexperienced group (16 students and 8 residents with <10 laparoscopic surgeries performed), performed three box trainer tasks validated for assessment of MIS psychomotor skills. Instrument movements were captured using the TrEndo tracking system, and nine motion analysis parameters (MAPs) were analyzed. The performance of the classifiers was measured by leave-one-out cross-validation using the scores obtained by the participants. Results The mean accuracy performances of the classifiers were 71 % (LDA), 78.2 % (SVM), and 71.7 % (ANFIS). No statistically significant differences in the performance were identified between the classifiers. Conclusions The three proposed classifiers showed good performance in the discrimination of skills, especially when information from all MAPs and tasks combined were considered. A correlation between the surgeons’ previous experience and their execution of the tasks could be ascertained from results. However, misclassifications across all the classifiers could imply the existence of other factors influencing psychomotor competence.

Face, content, and construct validity of the EndoViS training system for objective assessment of psychomotor skills of laparoscopic surgeons

Background The aim of this study is to present face, content, and constructs validity of the endoscopic orthogonal video system (EndoViS) training system and determines its efficiency as a training and objective assessment tool of the surgeons’ psychomotor skills. Methods Thirty-five surgeons and medical students participated in this study: 11 medical students, 19 residents, and 5 experts. All participants performed four basic skill tasks using conventional laparoscopic instruments and EndoViS training system. Subsequently, participants filled out a questionnaire regarding the design, realism, overall functionality, and its capabilities to train hand–eye coordination and depth perception, rated on a 5-point Likert scale. Motion data of the instruments were obtained by means of two webcams built into a laparoscopic physical trainer. To identify the surgical instruments in the images, colored markers were placed in each instrument. Thirteen motion-related metrics were used to assess laparoscopic performance of the participants. Statistical analysis of performance was made between novice, intermediate, and expert groups. Internal consistency of all metrics was analyzed with Cronbach’s α test. Results Overall scores about features of the EndoViS system were positives. Participants agreed with the usefulness of tasks and the training capacities of EndoViS system (score >4). Results presented significant differences in the execution of three skill tasks performed by participants. Seven metrics showed construct validity for assessment of performance with high consistency levels. Conclusions EndoViS training system has been successfully validated. Results showed that EndoViS was able to differentiate between participants of varying laparoscopic experience. This simulator is a useful and effective tool to objectively assess laparoscopic psychomotor skills of the surgeons.

Relevance of motion-related assessment metrics in laparoscopic surgery

INTRODUCTION: Motion metrics have become an important source of information when addressing the assessment of surgical expertise. However, their direct relationship with the different surgical skills has not been fully explored. The purpose of this study is to investigate the relevance of motion-related metrics in the evaluation processes of basic psychomotor laparoscopic skills, as well as their correlation with the different abilities sought to measure. METHODS: A framework for task definition and metric analysis is proposed. An explorative survey was first conducted with a board of experts to identify metrics to assess basic psychomotor skills. Based on the output of that survey, three novel tasks for surgical assessment were designed. Face and construct validation study was performed, with focus on motion-related metrics. Tasks were performed by 42 participants (16 novices, 22 residents and 4 experts). Movements of the laparoscopic instruments were registered with the TrEndo tracking system and analyzed. RESULTS: Time, path length and depth showed construct validity for all three tasks. Motion smoothness and idle time also showed validity for tasks involving bi-manual coordination and tasks requiring a more tactical approach respectively. Additionally, motion smoothness and average speed showed a high internal consistency, proving them to be the most task-independent of all the metrics analyzed. CONCLUSION: Motion metrics are complementary and valid for assessing basic psychomotor skills, and their relevance depends on the skill being evaluated. A larger clinical implementation, combined with quality performance information, will give more insight on the relevance of the results shown in this study.

Obtención de clones de alcornoque (Quercus suber L.) mediante embriogénesis somática: aplicación de medios líquidos y biorreactores en la sincronización de los procesos de desarrollo y maduración de los embriones

El alcornoque tiene un gran valor ambiental, como integrante de los ecosistemas forestales mediterráneos, e interés comercial por el valor de la bellota (alimentación del cerdo ibérico), el carbón, la madera y sobre todo por las aplicaciones industriales del corcho. Las posibilidades de mejora genética del alcornoque, como las de otras especies forestales, están limitadas por sus largos ciclos reproductivos y porque su propagación vegetativa mediante estaquillado solo es posible en estados muy juveniles. Por ello este sistema de propagación tiene muy poca, o ninguna, utilidad práctica en la mejora genética. La embriogénesis somática es la vía más apropiada para la clonación de muchas especies forestales y ha hecho posible el desarrollo a gran escala de plantaciones multivarietales de coníferas. En alcornoque es posible la regeneración completa de árboles adultos mediante embriogénesis somática. Con los protocolos actuales (en medio semisólido), los embriones se generan formando acúmulos y en la fase de multiplicación conviven embriones en distintos estados de desarrollo. Es un sistema asincrónico, con baja eficacia para la propagación en masa, que no elimina completamente las dificultades para el desarrollo de programas de mejora genética del alcornoque. En otras especies la utilización de medios líquidos ha mejorado: la sincronización, productividad de los cultivos, el manejo y reducido los costes de producción. Por ello el desarrollo de suspensiones embriogénicas de alcornoque se plantea como una vía para aumentar la eficacia de la propagación clonal a gran escala. En la presente tesis se desarrollan cultivos embriogénicos de alcornoque en medio líquido. El capítulo 3 aborda el establecimiento y mantenimiento de suspensiones, el capítulo 4 el desarrollo de una fase de proliferación en medio líquido y el capítulo 5 la utilización de sistemas de cultivo en medio líquido, estacionarios y de inmersión temporal, como vía para favorecer la maduración de los embriones somáticos. Para iniciar los cultivos en medio líquido se emplearon agregados de embriones tomados de la fase de proliferación en medio semisólido. Cuando estos agregados se inocularon directamente en medio líquido no se logró el establecimiento de las suspensiones. El establecimiento se consiguió empleando como inóculo las células y Resumen pequeños agregados embriogénicos, de tamaño comprendido entre 41 y 800 μm, desprendidas por agitación breve de los agregados de embriones. El mantenimiento se logró inoculando en baja densidad masas embriogénicas compactas de tamaño comprendido entre 0,8 y 1,2 mm. Estas suspensiones, muy heterogéneas, mantuvieron su capacidad de proliferación y de regeneración de embriones al menos durante diez subcultivos consecutivos. El protocolo de iniciación y mantenimiento, desarrollado inicialmente con un solo genotipo, fue eficaz cuando se probó sobre otros 11 genotipos de alcornoque. En la fase de proliferación se ensayaron tres tipos de envase y tres velocidades de agitación. La combinación envase × velocidad determinó el intercambio gaseoso, la disponibilidad de oxígeno y el estrés hidrodinámico. Los agregados embriogénicos de alcornoque crecieron incluso en condiciones de hipoxia no siendo la disponibilidad de oxígeno un factor limitante del crecimiento para tasas de trasferencia de oxígeno comprendidas entre 0,11 h-1 y 1,47 h-1. Por otra parte la producción de biomasa creció con el estrés hidrodinámico para valores de índice de cizalladura inferiores a 5 x 10-3 cm min-1. La mayor producción de biomasa se obtuvo con matraces Erlenmeyer de 100 ml y alta velocidad de agitación (160 rpm) mientras que la diferenciación de embriones se vio favorecida por bajas velocidades de agitación (60 rpm) asociadas con bajas disponibilidades de oxígeno. La posibilidad de madurar embriones de alcornoque en medio líquido se estudió utilizando sistemas de inmersión permanente y sistemas de inmersión temporal. En inmersión permanente no se diferenciaron embriones cotiledonares (posiblemente por hiperhidricidad). Los sistemas de inmersión temporal permitieron obtener embriones maduros en estado cotiledonar y capaces de regenerar plantas in vitro. Concentraciones de sacarosa superiores a 60 g l-1 y frecuencias de inmersión iguales o inferiores a una diaria, tuvieron efectos negativos para el desarrollo de los embriones somáticos. En los sistemas de inmersión temporal los parámetros físico-químicos del medio de cultivo se mantuvieron estables y no se observó ninguna limitación de nutrientes. No obstante, estos sistemas se vieron afectados por la evaporación que generó el flujo de aire necesario para desplazar el líquido en cada periodo de inmersión. Abstract ABSTRACT Cork oak is one of the most important tree species of the Mediterranean ecosystem. Besides its high environmental value has a great economic interest due to the sustainable production of acorns (to feed the Iberian pig) charcoal, timber and cork, which is a renewable natural product with various technological applications. As happens with other forest species, cork oak genetic improvement programs are limited by their long life cycles and because vegetative propagation by cuttings it´s only possible in very juvenile plants. Hence this propagation system is useless or has little practical use for breeding cork oak. Plant regeneration by somatic embryogenesis is the most suitable way for cloning many forest species, and it is the enabling technology which has allowed the establishment of large-scale conifer multi-varietal plantations. Clonal plant regeneration of mature cork oak trees can be achieved through somatic embryogenesis. Somatic embryos at different stages of development and forming clusters are produced during the multiplication phase with current protocols (using semisolid medium). This is an asynchronous low-efficient process not suitable for mass propagation, and therefore it does not solve the difficulties presented by cork oak breeding programs. Culture in liquid medium has been used with other species to improve: synchronization, yield, handling, and to reduce production costs. Thus the development of cork oak embryogenic suspension cultures is envisaged as a way to increase the efficiency of large scale clonal propagation. The thesis herein develops cork oak embryogenic cultures in liquid medium. In chapter 3 establishment and maintenance of suspension cultures are developed, chapter 4 studies proliferation phase in liquid medium and chapter 5 considers the use of different systems of culture in liquid medium, both stationary and temporary immersion, as a way to promote somatic embryos maturation. Clusters of embryos taken from proliferating cultures on semisolid medium were used to initiate the cultures in liquid medium. When these clusters were inoculated directly in liquid medium establishment of suspension cultures was not executed. However using, as initial inoculum, cells and cell aggregates with a size between 41 and 800 μm detached from these clusters of embryos, subjected to a brief shaking, suspension cultures could be established. Suspension maintenance was achieved by inoculating compact embryogenic Abstract clumps with a size between 0.8 and 1.2 mm at low density. The suspension cultures, very heterogeneous, retained both their proliferation and embryo regeneration capacity for at least ten consecutive subcultures. The initiation and maintenance protocol, initially developed with a single genotype, was effective when tested on 11 additional genotypes of cork oak. In proliferation phase three types of vessels and three different levels of agitation were assayed. The combination vessel × orbiting speed determined gas exchange, oxygen availability and hydrodynamic stress. Cork oak embryogenic aggregates grew even under hypoxia conditions; oxygen availability at transfer rates between 0.11 and 1.47 h-1 was not a limiting factor for growth. Furthermore the biomass production was increased with hydrodynamic stress when shear rate values were of less than 5 x 10-3 cm min-1. The highest biomass production was obtained with 100 ml Erlenmeyer flask and high stirring speed (160 rpm) while the differentiation of embryos was favored by low agitation speeds (60 rpm) associated with low oxygen availability. The possibility to mature cork oak somatic embryos in liquid medium was studied using both permanent immersion systems and temporary immersion systems. Cotyledonary embryos did not differentiate in permanent immersion conditions (probably due to hyperhydricity). Temporary immersion systems allowed obtaining mature cotyledonary embryos, which were able to regenerate plants in vitro. Sucrose concentrations above 60 g l-1 and immersion frequencies equal to or lower than one each 24 h had negative effects on somatic embryo development. Physicochemical parameters of the culture medium in temporary immersion systems were stable and showed no limitation of nutrients. However, these systems were affected by the evaporation generated by the airflow necessary to relocate the medium at each immersion period.

Caracterización de la agitación local y la respuesta oscilatoria de un buque mediante el uso de técnicas de visión artificial : aplicación al análisis de los umbrales operativos en líneas de atraque y amarre

El desarrollo de actividades de carga y descarga son parte de la esencia de la naturaleza funcional de un puerto, de las cuales derivan en gran medida los ingresos del mismo y la eficiencia de la cadena logística en su conjunto. Las oscilaciones en el interior de una dársena y en un línea de atraque disminuyen la calidad de la estancia de las embarcaciones en puerto, reducen el rendimiento de la estiba de los buques y solicitan y fatigan las estructuras y los cuerpos flotantes amarrados. Si los parámetros que definen la agitación local se aproximan a regiones de fallo 0 parada, el subsistema pierde rendimiento, fiabilidad y finalmente se paralizan las operaciones, produciéndose de este modo tiempos de inactividad. Estas paradas operativas conllevan pérdidas económicas para la terminal y, consecuentemente, para el puerto. Hoy día se dispone vastas redes de monitorización destinadas a la caracterización del medio físico en el entorno de los puertos. Paralelamente, las operaciones de manipulación de cargas en las terminales se están dirigiendo hacia modelos de automatización o semi automatización, que permiten no sólo la sistematización de procesos, sino también un profundo conocimiento del flujo de tareas. En este contexto hay un déficit de información sobre cómo afectan los diferentes forzadores del medio físico al rendimiento, la seguridad funcionalidad del proceso de manipulación de carga y descarga. Esto se debe en gran medida a la falta de registros dilatados en el tiempo que permitan correlacionar todos los aspectos mencionados de un modo particularizado para cada línea de atraque y amarre de un puerto. En esta tesis se desarrolla una metodología de vídeo monitorización no intrusiva y de bajo coste basada en la aplicación de técnicas "pixel tool' y la obtención de los parámetros extrínsecos de una observación monofocal. Con ello pretende poner en valor las infraestructuras de vídeo vigilancia de los puertos y de los laboratorios de experimentación a escala reducida, con el objeto de facilitar el estudio los umbrales operativos de las áreas de atraque y amarre. The development of loading and unloading activities is an essential part of he functional nature of a port, which derive largely from he same income and the efficiency of he supply chain as a whole. The oscillations inside a dock and a mooring line diminish he quality of the stay of vessels in port reducing the performance of the stowage of ship and asking and fatigued structures and moored floating bodies. If the parameters defining the local al agitation regions are close to areas of failure or shutdown, he subsystem looses performance, reliability and eventually paralyzes the operations, thereby producing downtime. These operational stops entail economic 1osses to the terminal and, consequently for the port. Today vast networks of monitoring, aimed at he characterization of the physical environment in the vicinity of he ports, are available. In parallel, the cargo handling operations at terminals are moving towards automation or semi-automation models that allow not only the systematization of processes, but also a deep understanding of he workflow. In this context, there is a lack of information about how the different forcing agents of the physical environment affect the performance and he functional safety of the loading and unloading process. This is due largely to the lack of spread-over-time records which would allow to correlate all aspects mentioned, specifically, for each berthing and mooring of a port. This thesis develops a methodology for non-intrusive and low cost monitoring video based on the application of "pixel tool" techniques and on obtaining the extrinsic parameters of a monofocal observation. It seeks an enhancement of the video monitoring infrastructure at ports and at experimental laboratories of reduced scale, in order to facilitate the study of operational thresholds berthing and mooring areas.

Riesgo del consumo de cannabis para la práctica físico-deportiva

El consumo de cannabis es uno de los principales problemas de salud pública, especialmente entre los jóvenes, dadas sus prevalencias de consumo. Esta droga puede provocar alteraciones psicomotrices en el individuo, por ello, el presente trabajo de investigación tiene como objetivo principal comprobar si el consumo de cannabis supone un riesgo añadido a la práctica físico-deportiva. Para alcanzar dicho objetivo, se dividió el trabajo en dos estudios interrelacionados entre sí. En el primer estudio o Estudio 1, “Repercusión del consumo de cannabis en las clases de Educación Física”, se realizó un cuestionario autoadministrado, dirigido al profesorado de Educación Física de la Comunidad de Madrid, con el fin de recoger su opinión sobre la posible presencia de consumidores de cannabis en sus aulas. El segundo estudio o Estudio 2, “Influencia del cannabis en el equilibrio, la coordinación, el tiempo de respuesta y la percepción temporal”, consistió en la realización de una serie de pruebas por parte de un grupo control y un grupo de consumidores de cannabis, para así cuantificar y cualificar los efectos de la inhalación de esta droga en la condición motriz. La encuesta que se utilizó para el Estudio 1, “El cannabis, los adolescentes y la práctica físico-deportiva”, pasó primero por una fase de elaboración, corrección y validación, y después, utilizándose la técnica de muestreo aleatorio por conglomerados, fue distribuido por los centros públicos de enseñanzas secundarias de la Comunidad de Madrid seleccionados. Se obtuvo respuesta de 93 profesores de Educación Física pertenecientes a 76 centros, es decir el 22,35% de los centros de la comunidad. Por último, se procedió al análisis, discusión y difusión de los resultados. En el Estudio 2, primero se procedió al diseño de la batería de pruebas a realizar, compuesta por 7 tests con 16 variables que valoraron el equilibrio, la coordinación, el tiempo de respuesta y la percepción temporal de los sujetos. Después se procedió a la selección de la muestra, formada por dos grupos de voluntarios de entre 18 y 30 años: un grupo control de 52 participantes, y un grupo de consumidores de cannabis, de 28 participantes con un hábito de consumo diferente. Los sujetos del grupo de consumidores realizaron las pruebas 20 minutos después de la inhalación de cannabis. Tras la realización de las pruebas por parte de todos los sujetos se procedió al análisis de los resultados obtenidos, segmentándolos por sexo y apareando las variables para evitar posibles efectos confundidores. Los resultados del primer estudio indicaron que el 75,27% del profesorado ha pensado alguna vez que sus alumnos consumían cannabis antes del inicio de su clase, hecho asociado a alteraciones psicológicas (90,63%) y cognitivo-conductuales (68,75%); no siendo factores influyentes el sexo o la experiencia docente de los encuestados (p>0,05). El segundo estudio aportó como principal resultado el empeoramiento de la percepción temporal y la conducta motriz de los sujetos que consumieron cannabis, en especial de su equilibrio y su coordinación (p<0,05). También indicó que hubo ligeras diferencias en función del hábito de consumo de esta droga, si bien los resultados se vieron influenciados por el tamaño de la muestra. Las conclusiones extraídas mostraron que el profesorado de Educación Física percibe la presencia de consumidores de cannabis en sus aulas, utiliza el diálogo con el alumnado como herramienta de actuación, y conoce la influencia negativa que tiene el consumo de esta droga en el rendimiento motor. También demuestran que el cannabis supone un riesgo añadido a la práctica físico-deportiva, debido al empeoramiento de la conducta motriz, en especial de la coordinación y el equilibrio. Además, las diferencias surgidas en base al hábito de consumo no son concluyentes pero si apuntan a que el consumo esporádico puede conllevar un peor rendimiento motor que el consumo habitual. ABSTRACT Cannabis use is one of the most important concerns of public health, particularly among young people, given their use prevalence. This drug can produce psychomotor alterations in the subject; therefore, this research work has as main aim to check if cannabis use is an added risk for physical and sporting practices. In order to achieve the objectives pursued, work was divided in two interrelated studies. First study or Study 1, “Cannabis use impact in Physical Education lessons”, a self-managed questionnaire addressed to Physical Education teachers in Community of Madrid was carried out, in order to gather the opinion about the possible existence of Cannabis users in their classes. Second Study or Study 2, “Cannabis influence in balance, coordination, reaction time and temporal perception”, consisted in a series of tests performed by a control group and a cannabis users group, in order to quantify and qualify the effects of inhalation of this drug in the motor skills. The questionnaire used for Study 1, “Cannabis, teenagers, and physical and sporting practices”, firstly went through an elaboration y correction phase, and then, using cluster sampling technique, was distributed within selected Secondary Education Centres from Community of Madrid. Response from 93 Physical Education teachers from 76 High Schools were obtained, this is 22,35% of total High Schools in Madrid. Finally, analysis, discussion and dissemination of results were held. In Study 2, first of all design of tests to be made was done, consisting on 7 different test with 16 variables that measured balance, coordination, response time and temporal perception in subjects. After that, sample selection was performed, comprised of 2 volunteers groups of ages between 18-30 years: a control group of 52 participants, and a Cannabis Users Group of 28 participants with different consumption habits. Users group subjects carried out tests 20 minutes after cannabis inhalation. After tests were performed by all subjects, results were analyzed and segmented by sex and matching variables in order to avoid confusion effects. First study results brought out that 75,27% of teachers have ever thought that their pupils used cannabis before getting into class, this fact is linked to psychological alterations (90,63%) and cognitive-behavioral (68,75%); not being influencing factors sex or teaching experience of the surveyed (p>0,05). Second study provided as most significant result the worsening of temporal perception and motor tasks behavior in subjects that had used cannabis, especially in regards of their balance and coordination (p<0,05). Also resulted that there were slight differences in consumption habits, although results were influenced by the sample size. Conclusions drawn showed that Physical Education teachers notice the presence of cannabis users in lessons, use dialogue with pupils as an action tool and acknowledge the negative influence this drug has in motor tasks performance. Also, it is shown that cannabis is an additional risk for physical and sporting practices due to deteriorating in motor skills, particularly in coordination and balance. In addition, the differences that may arise depending on the consumption habits are not conclusive, but they suggest that the sporadic drug use may be related to a worse motor performance than usual consumption.

Optimización de la agitación de un digestor anaerobio mediante mecánica de fluidos computacional

Debido al gran interés existente en el ahorro y recuperación de energía, y en el deseo de obtener productos que permitan usos beneficiosos del fango procedente de la depuración del agua residual, la digestión anaerobia es el proceso de estabilización de uso más extendido. El tiempo de retención de sólidos es un factor clave en el proceso de digestión anaerobia. En base al tiempo de retención de sólidos, se dimensiona el volumen de los digestores anaerobios para así obtener la reducción de materia orgánica deseada, con la correspondiente producción de biogás. La geometría del digestor y su sistema de agitación deben ser adecuados para alcanzar el tiempo de retención de sólidos de diseño. Los primeros trabajos sobre la agitación de los digestores realizaban únicamente experimentos con trazadores y otros métodos de medición. En otros casos, la mezcla era evaluada mediante la producción de biogás. Estas técnicas tenían el gran hándicap de no conocer lo que sucedía realmente dentro del digestor y sólo daban una idea aproximada de su funcionamiento. Mediante aplicación de la mecánica de fluidos computacional (CFD) es posible conocer con detalle las características del fluido objeto de estudio y, por lo tanto, simular perfectamente el movimiento del fango de un digestor anaerobio. En esta tesis se han simulado mediante CFD diferentes digestores a escala real (unos 2000 m3 de volumen) agitados con bomba/s de recirculación para alcanzar los siguientes objetivos: establecer la influencia de la relación entre el diámetro y la altura, de la pendiente de la solera, del número de bombas y del caudal de recirculación en dichos digestores, definir el campo de velocidades en la masa de fango y realizar un análisis energético y económico. Así, es posible conocer mejor cómo funciona el sistema de agitación de un digestor anaerobio a escala real equipado con bomba/s de recirculación. Los resultados obtenidos muestran que una relación diámetro/altura del digestor por encima de 1 empeora la agitación del mismo y que la pendiente en la solera del digestor favorece que la masa de fango esté mejor mezclada, siendo más determinante la esbeltez del tanque que la pendiente de su solera. No obstante, también es necesario elegir adecuadamente los parámetros de diseño del sistema de agitación, en este caso el caudal de recirculación de fango, para obtener una agitación completa sin apenas zonas muertas. En el caso de un digestor con una geometría inadecuada es posible mejorar su agitación aumentando el número de bombas de recirculación y el caudal de las mismas, pero no se llegará a alcanzar una agitación total de la masa de fango debido a su mal diseño original. Anaerobic digestion is the process for waste water treatment sludge stabilization of more widespread use due to the huge interest in saving and recovering energy and the wish to obtain products that allow beneficial uses for the sludge. The solids retention time is a key factor in the anaerobic digestion. Based on the solids retention time, volume anaerobic digester is sized to obtain the desired reduction in organic matter, with the corresponding production of biogas. The geometry of the digester and the stirring system should be adequate to achieve the design solid retention time. Early works on digesters stirring just performed tracer experiments and other measurement methods. In other cases, mixing was evaluated by biogas production. These techniques had the great handicap of not knowing what really happened inside the digester and they only gave a rough idea of its operation. By application of computational fluid dynamics (CFD), it is possible to know in detail the characteristics of the fluid under study and, therefore, simulate perfectly the sludge movement of an anaerobic digester. Different full-scale digesters (about 2000 m3 of volume) agitated with pump/s recirculation have been simulated by CFD in this thesis to achieve the following objectives: to establish the influence of the relationship between the diameter and height, the slope of the bottom, the number of pumps and the recirculation flow in such digesters, to define the velocity field in the mass of sludge and carry out an energy and economic analysis. Thus, it is possible to understand better how the agitation system of a full-scale anaerobic digester equipped with pump/s recirculation works. The results achieved show that a diameter/height ratio of the digester above 1 worsens its stirring and that the slope of the digester bottom favors that the mass of sludge is better mixed, being more decisive the tank slenderness than the slope of its bottom. However, it is also necessary to select properly the design parameters of the agitation system, in this case the sludge recirculation flow rate, for a complete agitation with little dead zones. In the case of a digester with inadequate geometry, its agitation can be improved by increasing the number of recirculation pumps and flow of them, but it will not reach a full agitation of the mass of sludge because of the poor original design.

Desarrollo de un sistema automático de detección de disgrafías en niños de 4 a 5 años

La escritura es una actividad psicomotora muy importante en el desarrollo infantil. Tanto es así que su correcto aprendizaje condicionará el futuro de una persona, pues estará presente en todo tipo de situaciones cotidianas. La disgrafía es el término utilizado para referirse a los problemas relacionados con la escritura, y se manifiesta cuando la escritura de un determinado sujeto es ilegible o lenta como resultado de un aprendizaje fallido de los procesos motores de la escritura. Estos problemas intentan resolverse durante el desarrollo infantil mediante diferentes pruebas que miden las capacidades visomotoras de los niños basándose en criterios de forma (número y posición correcta de trazos). Sin embargo, a lo largo de los años estos criterios han demostrado no ser totalmente precisos en la detección prematura de posibles casos de disgrafía. Por ello, en este proyecto se ha desarrollado una aplicación que ayuda a ampliar la fiabilidad de los test actuales, utilizando un criterio cinemático. Esta aplicación, desarrollada para una tableta Android, muestra al niño una serie de figuras que él debe copiar en la tableta haciendo uso de un lápiz óptico. Los trazos registrados por la aplicación son analizados para valorar aspectos como la fluidez, continuidad y regularidad, ampliando así la fiabilidad de los test actuales, lo que permite desechar falsos positivos y detectar irregularidades que antes no podían ser detectadas. La aplicación desarrollada ha sido validada con un total de ocho niños de cuatro años y siete meses de media de edad, confirmando que cumple con las expectativas planteadas. ABSTRACT. Writing is a very important psychomotor activity in child development because it will be present in all kinds of everyday situations; therefore its proper learning will determine the future of the individual. Dysgraphia is the term used to refer to the problems related to writing, and it takes place when a particular person’s writing is unreadable or slow-moving as a result of a failed learning of writing motor processes. These problems are usually detected by different tests that measure children’s visual motor abilities based on shape criteria (correct number and position of strikes). However, over the years these criteria haven’t proved to be completely accurate in the early detection of possible cases of dysgraphia. Therefore, in this project is presented an application that extends the reliability of current test, using a kinematic approach. This application, developed for an Android tablet, displays a series of figures that the child must copy to the tablet by using a stylus. Strokes recorded by the application are then analyzed to assess aspects such as fluidity, continuity and regularity, expanding the reliability of the current test, discarding false positives created by the conventional criteria and detecting irregularities that previously could not be detected. The developed application has been validated with a total of eight children about four years and seven months in average age, confirming that the application fulfills the initial expectations.