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.

Modelo de política pública de I+D+i, para liderar y coordinar el sistema regional de innovación en apoyo a las PYMIS. Caso de estudio el estado Bolívar, Venezuela

This proposed project aims to design a Model of Public Policy for R&D &I to lead, coordinate and direct the Regional Innovation System (RIS) in support of the SMI sector. The case study of the Bolivar State, Venezuela, is analyzed. The project represents a no- experimental research of projective type and is divided into two parts. In the first one, the current state of the SRI and its relationship with the SMIs is diagnosed, evaluated and described. In the second part, a model of public policy for R&D&I is designed and proposed in order to strengthen the capacity for innovation in SMIs.

Articulación del sistema regional de onnovación para desarrollar capacidad de I+D+i

El sector de las Pequeñas y Medianas Industrias(PYMIS) constituye una proporción importante del tejido industrial del estado Bolívar. Sin embargo, en los últimos años se ha visto afectada su competitividad por la deficiencia en la capacidad de I+D+i, para la creación de nuevos productos que responda a las necesidades del mercado. El presente proyecto se planteó como objetivo evaluar, caracterizar y diseñar un modelo para articular el Sistema Regional de Innovación (SRI), para el desarrollo de la capacidad de la I+D+i, en el sector de las PYMIS del estado Bolívar, Venezuela. El estudio se realizó a través de una investigación no experimental, tipo evaluativo y aplicado. La recolección de la información se realizo mediante la aplicación de tres tipos de encuestas, destinadas a los siguientes actores que conforman el SRI: PYMIS, centros y laboratorios de I+D e instituciones financieras públicas y privadas. En cada una se consideró el rol y las relaciones con la institución gubernamental que define la política de ciencia y tecnología. Entre las conclusiones se tiene que la falta de articulación de los actores del SRI, afecta al aprovechamiento de los recursos, a la infraestructura existente para la I+D+i, y a la respuesta por parte de las PYMIS a las oportunidades del mercado.

Diseño de una metodología para la aplicación y evaluación de políticas de innovación para el desarrollo de las capacidades de la I+D+i en las pequeñas y medianas industrias

El presente trabajo tiene por objetivo el diseño de una Metodología para la Aplicación y Evaluación de Políticas de Innovación para el Desarrollo de las Capacidades de la I+D+i en las Pequeñas y Medianas Industrias (PYMIS), caso de las PYMIS del estado Bolívar, Venezuela. El diseño de la metodología se inicia con el análisis de las capacidades de I+D+i en las PYMIS y de su relación con los actores del SRI, posteriormente se estudió los diferentes modelos que describen el funcionamiento de los sistemas de innovación, obteniendo el diseño de la metodología de siete etapas, para la aplicación y evaluación de políticas de innovación.

Sistema regional de innovación para el fortalecimiento de la I+D+i, en el sector industrial del estado Bolívar, Venezuela

El Estado Bolívar cuenta con excelentes recursos de materias primas tales como el mineral de hierro y bauxita, y de profesionales especializados, los cuales representan oportunidades para mejorar las ventajas competitivas de una de las zonas industriales más grande de Latinoamérica. Para aprovechar tales ventajas se debe cambiar el modelos de política industrial actual de exportar en el mayor porcentaje productos primarios y semielaborados a través de las empresa básicas instaladas en la zona, a otro modelo económico basado en política de innovación para la diversificación de nuevos productos, soportados en la transformación de las Pequeñas y Medianas Industrias (PYMIS) del estado Bolívar, consolidando las empresas en organizaciones innovadoras que generen, aporten o adapten los conocimientos científicos y tecnológicos, obteniendo nuevos productos o mejoras de procesos con un alto contenido de valor agregado. Estos cambios avanzarán en la medida que se logren articular e integrar los actores que forman parte del Sistema Regional de Innovación (SRI). El presente proyecto se planteo como objetivo diseñar estrategias para desarrollar y articular las capacidades de los actores del SRI para el fortalecimiento de la I+D+i en el sector industrial de las cadenas siderúrgicas y del aluminio del estado Bolívar.

Diseño de una Metodología de Política Pública de I+D+i para el Desarrollo de la Capacidad de la Innovación en el Sector PYMIS del estado Bolívar, Venezuela

El estado Bolívar con una superficie de 238.000 Km2 se encuentra ubicado al SE de Venezuela y su capital es Ciudad Bolívar. Ocupa el 26,24% de la superficie del territorio nacional. Ciudad Guayana es la principal región del desarrollo económico del estado siendo sede de las empresas básicas de los sectores siderúrgicos y del aluminio que se encargan de la extracción, procesamiento y transformación del mineral de hierro y de la transformación de la bauxita en aluminio primario. Además, cuenta con un gran potencial hidroeléctrico, garantizando el suministro de energía eléctrica para el funcionamiento de las empresas básicas, para el parque industrial de la región, así como para el desarrollo industrial, económico y social de la nación. Con relación al sector de la industria del mineral de hierro y del aluminio, las empresas destinan más del 60 por ciento de su producción al mercado internacional. A pesar de que el sector de las Pequeñas y Medianas Industrias (PYMIS) del estado Bolívar cuenta con un mercado cercano y seguro, no se le ha propiciado un desarrollo integral en términos de orientar sus esfuerzos en innovar en nuevos productos o mejoras de procesos. Debido a la falta del personal de investigación calificado, la escasa vinculación con centros de investigación, la baja inversión en investigación, desarrollo tecnológico e innovación (I+D+i), la ausencia de la aplicación de una política pública de I+D+i y la desarticulación de los miembros del Sistema Regional de Innovación (SRI), constituyen los principales obstáculos para generar bienes y servicios con un alto valor agregado. Esta situación desequilibra y hace ineficiente el funcionamiento del SRI. La baja capacidad de las PYMIS del estado Bolívar en I+D+i, es una situación que impide generar por si sola nuevos productos o procesos para satisfacer las demandas del mercado regional. Por lo tanto, se requiere de la intervención y participación activa de la institución gubernamental responsable del diseño y aplicación de una política pública de I+D+i para dinamizar la capacidad de innovación en las PYMIS, en su articulación y vinculación con los miembros del SRI. xiii El presente proyecto se planteó como objetivo diseñar una metodología de política pública de I+D+i para liderar, coordinar y direccionar el SRI del estado Bolívar, para el desarrollo de la capacidad de innovación en el sector industrial y específicamente en las PYMIS. La presente tesis representa una investigación no experimental de tipo proyectivo que analiza la situación actual del Sistema Regional de Innovación del estado Bolívar. El análisis de los resultados se ha dividido en tres fases. En la primera se realizan diagnósticos por medio de encuestas de las PYMIS en materia de I+D+i, de los centros y laboratorios de investigación pertenecientes a las universidades de la región en el área de Materiales y de los sectores financieros público y privado. En dichas encuestas se evalúa el nivel de integración con los entes gubernamentales que definen y administran la política pública de I+D+i. En la segunda fase, con el diagnóstico y procesamiento de los resultados de la primera fase, se procede a desarrollar un análisis de las fortalezas, oportunidades, debilidades y amenazas (FODA) del Sistema Regional de Innovación, permitiendo comprender la situación actual de la relación y vinculación de las PYMIS con los centros de investigación, instituciones financieras y entes gubernamentales. Con la problemática detectada, resultó necesario el diseño de estrategias y un modelo de gestión de política pública de I+D+i para la articulación de los miembros del SRI, para el apoyo de las PYMIS. En la tercera fase se diseña la metodología de política pública de I+D+i para fortalecer la innovación en las PYMIS. La metodología se representa a través de un modelo propuesto que se relaciona con las teorías de los procesos de innovación, con los modelos de sistemas de innovación y con las reflexiones y recomendaciones hechas por diferentes investigadores e instituciones de cooperación internacional referentes a la aplicación de políticas públicas de I+D+i para dinamizar la capacidad de innovación en el sector industrial. La metodología diseñada es comparada con diferentes modelos de aplicación de política pública de I+D+i. Cada modelo se representa en una figura y se analiza su xiv situación presente y la función que desempeña el ente gubernamental en la aplicación del enfoque de política pública de I+D+i. El diseño de la metodología de política pública de I+D+i propuesta aportará nuevos conocimientos y podrá ser aplicado para apoyar el progreso de la I+D+i en las PYMIS de la región, como caso de estudio, con el fin de impulsar una economía más competitiva y reducir el grado de dependencia tecnológica. La metodología una vez evaluada podrá ser empleada en el contexto de la gran industria y en otras regiones de Venezuela y además, puede aplicarse en otros países con características similares en su tejido industrial. En la tesis doctoral se concluye que el desarrollo de la capacidad de innovación en las PYMIS depende del diseño y aplicación de la política pública de I+D+i como elemento dinamizador y articulador del SRI del estado Bolívar. xv ABSTRACT The Bolivar state with an area of 238,000 km2 is located in the SE of Venezuela and its capital is Ciudad Bolivar. It occupies a surface which is 26.24% of the national territory. Ciudad Guayana is the main area of the state's economic development and the location of the corporate headquarters of the basic steel and aluminum sectors that are responsible for the extraction, processing and transformation of iron ore and bauxite processing for primary aluminum. It also has a great hydroelectric potential, ensuring the supply of electricity for the operation of enterprises, for the regional industrial park as well as for the industrial, economic and social development of the nation. With regard to the iron ore and aluminum industry, companies allocate more than 60 percent of their production to the international market. Although the sector of Small and Medium Industries (SMIs) of the Bolivar state has a secure market, it has not been led to an integral development in terms of targeting its efforts on innovating new products or improving processes. Due to the lack of qualified research staff, poor links with research centers, low investment in research, technological development and innovation (R & D & I), the absence of the implementation of a public policy for R & D & I and the dismantling of the members of the Regional Innovation System (RIS), are the main obstacles to generate goods and services with high added value. This situation makes the RIS unbalanced and inefficient. The low capacity of Bolivar state’s SMIs in R & D & I, is a situation that cannot generate by itself new products or processes to meet regional market demands. Therefore, it requires the active involvement and participation of the government institution responsible for the design and implementation of R & D & I public policy to boost the innovation capacity in SMIs, through the connection and integration with members of the RIS. This project is intended to design a methodology aimed at public policy for R & D & I to lead, coordinate and direct the RIS of Bolivar state, for the development of innovation capacity in the industrial sector and specifically in the SMIs. xvi This thesis is an experimental investigation of projective type which analyzes the current situation of the Regional Innovation System of the Bolivar state. The analysis of the results is divided into three phases. In the first one, a diagnosis is performed through surveys of SMIs in R & D & I centers and research laboratories belonging to the universities of the region in the area of materials and public and private financial sectors. In such surveys the level of integration with government agencies that define and manage the public policy of R & D & I is assessed. In the second phase, with the diagnosis and processing of the results of the first phase, we proceed to develop an analysis of the strengths, weaknesses, opportunities and threats (SWOT) of the Regional Innovation System, allowing the comprehension of the current status of the relationship of SMIs with research centers, financial institutions and government agencies. With the problems identified it was necessary to design strategies and a model of public policy management of R & D & I for the articulation of the members of the RIS, to support the SMIs. In the third phase a public policy methodology for R & D & I is designed in order to strengthen innovation in SMIs. The methodology is shown through a proposed model that relates to the theories of the innovation process, with models of innovation systems and with the discussions and recommendations made by different researchers and institutions of international cooperation concerning the implementation of policies public for R & D & I to boost innovation capacity in the industrial sector. The methodology designed is compared with different models of public policy implementation for R & D & I. Each model is represented in a figure and its current situation and the role of the government agency in the implementation of the public policy approach to R & D & I is analyzed. The design of the proposed public policy methodology for R & D & I will provide new knowledge and can be applied to support the progress of R & D & I in the region’s SMIs, as a case study, in order to boost a more competitive economy and reduce the degree of technological dependence. After being evaluated the methodology can be used in the context of big industry and in other regions of Venezuela and can also be applied in other countries with similar characteristics in their industrial structure. xvii The thesis concludes that the development of the innovation capacity in SMIs depends on the design and implementation of the public policy for R & D & I as a catalyst and coordination mechanism of the Regional Innovation System of the Bolivar state.

Import of DNA into mammalian nuclei by proteins originating from a plant pathogenic bacterium

Import of DNA into mammalian nuclei is generally inefficient. Therefore, one of the current challenges in human gene therapy is the development of efficient DNA delivery systems. Here we tested whether bacterial proteins could be used to target DNA to mammalian cells. Agrobacterium tumefaciens, a plant pathogen, efficiently transfers DNA as a nucleoprotein complex to plant cells. Agrobacterium-mediated T-DNA transfer to plant cells is the only known example for interkingdom DNA transfer and is widely used for plant transformation. Agrobacterium virulence proteins VirD2 and VirE2 perform important functions in this process. We reconstituted complexes consisting of the bacterial virulence proteins VirD2, VirE2, and single-stranded DNA (ssDNA) in vitro. These complexes were tested for import into HeLa cell nuclei. Import of ssDNA required both VirD2 and VirE2 proteins. A VirD2 mutant lacking its C-terminal nuclear localization signal was deficient in import of the ssDNA–protein complexes into nuclei. Import of VirD2–ssDNA–VirE2 complexes was fast and efficient, and was shown to depended on importin α, Ran, and an energy source. We report here that the bacterium-derived and plant-adapted protein–DNA complex, made in vitro, can be efficiently imported into mammalian nuclei following the classical importin-dependent nuclear import pathway. This demonstrates the potential of our approach to enhance gene transfer to animal cells.

Electrophysiological characterization of specific interactions between bacterial sensory rhodopsins and their transducers

The halobacterial phototaxis receptors sensory rhodopsin I and II (SRI, SRII) enable the bacteria to seek optimal light conditions for ion pumping by bacteriorhodopsin and/or halorhodopsin. The incoming signal is transferred across the plasma membrane by means of receptor-specific transducer proteins that bind tightly to their corresponding photoreceptors. To investigate the receptor/transducer interaction, advantage is taken of the observation that both SRI and SRII can function as proton pumps. SRI from Halobacterium salinarum, which triggers the positive phototaxis, the photophobic receptor SRII from Natronobacterium pharaonis (pSRII), as well as the mutant pSRII-F86D were expressed in Xenopus oocytes. Voltage-clamp studies confirm that SRI and pSRII function as light-driven, outwardly directed proton pumps with a much stronger voltage dependence than the ion pumps bacteriorhodopsin and halorhodopsin. Coexpression of SRI and pSRII-F86D with their corresponding transducers suppresses the proton transport, revealing a tight binding and specific interaction of the two proteins. These latter results may be exploited to further analyze the binding interaction of the photoreceptors with their downstream effectors.

The primary structures of the Archaeon Halobacterium salinarium blue light receptor sensory rhodopsin II and its transducer, a methyl-accepting protein.

Recently, a large family of transducer proteins in the Archaeon Halobacterium salinarium was identified. On the basis of the comparison of the predicted structural domains of these transducers, three distinct subfamilies of transducers were proposed. Here we report isolation, complete gene sequences, and analysis of the encoded primary structures of transducer gene htrII, a member of family B, and its blue light receptor gene (sopII) of sensory rhodopsin II (SRII). The start codon ATG of the 714-bp sopII gene is one nucleotide beyond the termination codon TGA of the 2298-bp htrII gene. The deduced protein sequence of HtrII predicts a eubacterial chemotaxis transducer type with two hydrophobic membrane-spanning segments connecting sizable domains in the periplasm and cytoplasm. HtrII has a common feature with HtrI, the sensory rhodopsin I transducer; like HtrI, HtrII possesses a hydrophilic loop structure just after the second transmembrane segment. The C-terminal 299 residues (765 amino acid residues total) of HtrII show strong homology to the signaling and methylation domain of eubacterial transducer Tsr. The hydropathy plot of the primary structure of SRII indicates seven membrane-spanning alpha-helical segments, a characteristic feature of retinylidene proteins ("rhodopsins") from a widespread family of photoactive pigments. SRII shows high identity with SRI (42%), bacteriorhodopsin (BR) (32%), and halorhodopsin (24%). The crucial positions for retinal binding sites in these proteins are nearly identical, with the exception of Met-118 (numbering according to the mature BR sequence), which is replaced by Val in SRII. In BR, residues Asp-85 and Asp-96 are crucial in proton pumping. In SRII, the position corresponding to Asp-85 in BR is conserved, but the corresponding position of Asp-96 is replaced by an aromatic Tyr. Coexpression of the htrII and sopII genes restores SRII phototaxis to a mutant (Pho81) that contains a deletion in the htrI/sopI and insertion in htrII/sopII regions. This paper describes the first example that both HtrI and HtrII exist in the same halobacterial cell, confirming that different sensory rhodopsins SRI and SRII in the same organism have their own distinct transducers.

Protonatable residues at the cytoplasmic end of transmembrane helix-2 in the signal transducer HtrI control photochemistry and function of sensory rhodopsin I.

Neutral residue replacements were made of 21 acidic and basic residues within the N-terminal half of the Halobacterium salinarium signal transducer HtrI [the halobacterial transducer for sensory rhodopsin I (SRI)] by site-specific mutagenesis. The replacements are all within the region of HtrI that we previously concluded from deletion analysis to contain sites of interaction with the phototaxis receptor SRI. Immunoblotting shows plasmid expression of the htrI-sopI operon containing the mutations produces SRI and mutant HtrI in cells at near wild-type levels. Six of the HtrI mutations perturb photochemical kinetics of SRI and one reverses the phototaxis response. Substitution with neutral amino acids of Asp-86, Glu-87, and Glu-108 accelerate, and of Arg-70, Arg-84, and Arg-99 retard, the SRI photocycle. Opposite effects on photocycle rate cancel in double mutants containing one replaced acidic and one replaced basic residue. Laser flash spectroscopy shows the kinetic perturbations are due to alteration of the rate of reprotonation of the retinylidene Schiff base. All of these mutations permit normal attractant and repellent signaling. On the other hand, the substitution of Glu-56 with the isosteric glutamine converts the normally attractant effect of orange light to a repellent signal in vivo at neutral pH (inverted signaling). Low pH corrects the inversion due to Glu-56 -> Gln and the apparent pK of the inversion is increased when arginine is substituted at position 56. The results indicate that the cytoplasmic end of transmembrane helix-2 and the initial part of the cytoplasmic domain contain interaction sites with SRI. To explain these and previous results, we propose a model in which (i) the HtrI region identified here forms part of an electrostatic bonding network that extends through the SRI protein and includes its photoactive site; (ii) alteration of this network by photoisomerization-induced Schiff base deprotonation and reprotonation shifts HtrI between attractant and repellent conformations; and (iii) HtrI mutations and extracellular pH alter the equilibrium ratios of these conformations.