Design, development and validation of a multi-step plasma-based strategy for the direct functionalization of L605 cobalt chromium alloy for the grafting of bioactive molecules and its application in cardiovascular devices

Les maladies cardio-vasculaires sont la principale cause de mortalité dans le monde. Parmi celles-ci, et une des plus importantes, se trouve l'athérosclérose. Cette maladie se traduit par la formation d'une plaque sur les parois artérielles réduisant alors le diamètre luminal. La plaque d'athérome entrave la circulation du sang et peut se compliquer par la formation d'un thrombus artérielle pouvant provoquer un infarctus du myocarde. Une intervention capable de rétablir le flux (recanalisation) est alors nécessaire. Dans le cas des artères coronaires, l'intervention coronaire est percutanée (ICP) et consiste à amener et déployer jusqu'au site malade, une endoprothèse, appelé aussi stent. Un stent est un petit treillis métallique tubulaire qui permet de rouvrir la lumière de l'artère et de rétablir la circulation sanguine. Il sert aussi de support à l'artère malade pour empêcher son affaissement. Cependant, après implantation, certaines complications sont induites, telle que la resténose intra-stent (ISR) qui se caractérise par la réduction de la lumière de l'artère, reconduisant les problèmes créés par la plaque d'athérome. Ce phénomène est essentiellement dû à une prolifération excessive des cellules musculaires lisses, et qui résulte d'une lésion de l'endothélium lors de l'implantation. Afin de limiter cette complication, la première approche a été de changer les matériaux utilisés pour ces endoprothèses. Les principaux alliages utilisés pour fabriquer des stents sont l'acier inoxydable, les alliages de nitinol et ceux de chrome cobalt, plus particulièrement le L605. Ce dernier, dû à ses propriétés mécaniques, permet la fabrication de dispositifs plus minces, donc moins de métaux présents dans le corps humain, et a démontré induire moins de complications cliniques. Néanmoins, malgré la diminution des complications par rapport aux autres alliages, les endoprothèses nues en L605 ne s'intègrent que peu ou pas du tout dans le tissu artériel de l'hôte. Pour répondre aux exigences biologiques et cliniques, l'idéal serait d'avoir un dispositif qui favoriserait le recouvrement du dispositif par l'endothélium, ou « endothélialisation » et qui aurait un faible potentiel thrombotique et inflammatoire. L'approche couramment utilisée pour répondre à ces critères est de recourir à des dispositifs qui libèrent des médicaments anti-inflammatoires. Pour ce faire, il faut recouvrir les dispositifs métalliques avec des revêtements à base de polymères, en tant que couche intermédiaire, fonctionnalisée ensuite par des molécules bioactives. Toutefois, les dépôts de ces couches polymériques impliquent l'utilisation de chimie en solution incluant des solvants organiques. En outre, ces dernières démontrent avoir une faible adhésion au substrat métallique, dû au procédé utilisé, mais aussi un manque de cohésion. Lors de la procédure d'implantation, les stents subissent une déformation plastique, comme ces revêtements manquent de résistance, ils ont tendance à fissurer ou délaminer. Ce projet de recherche s'insère donc dans cette problématique générale, et propose une nouvelle approche qui permettrait d'éviter ce revêtement polymérique, tout en apportant les propriétés biologiques recherchées. Pour ce faire, la modification de surface proposée implique la fonctionnalisation directe des surfaces métalliques par un procédé plasma. Ce procédé permet de ne pas modifier les propriétés de cœur du matériau, et de créer des groupes fonctionnels en surface, ici des groupes amine réactifs (NH₂), qui servent de points d'ancrage pour le greffage ultérieur des molécules bioactives d'intérêt. En résumé, ce procédé original peut être divisé en 3 parties principales : a) préparation de la surface, b) fonctionnalisation par plasma et c) greffage de molécules bioactives. Tout au long de ce projet de recherche, l'optimisation de chaque partie a été réalisée en vue d'obtenir les propriétés adéquates et nécessaires pour l'application cardiovasculaire visée. Concernant la partie a), c'est-à-dire la préparation de la surface, les traitements suivants ont été testés : électropolissage, traitements thermiques et implantation ionique par immersion dans un plasma. Ces modifications ont été optimisées en vue d'obtenir une couche d'oxyde stable sous déformation présentant la meilleure résistance possible à la corrosion, tout en démontrant la plus haute efficacité d'amination directe par plasma pour la partie b). Enfin, en ce qui concerne le bloc c), le greffage de molécule bioactive, deux bras de liaison différents ont été étudiés pour évaluer leur impact sur la conformation et la performance biologique. Cette étude a été effectuée avec un peptide bioactif dérivé de la molécule d'adhésion des cellules endothéliales et des plaquettes (PECAM-1 ou CD31), en raison de ses propriétés anti-inflammatoire, anti-thrombotique et pro-endothélialisation. Les 2 bras d'ancrage testés sont un à chaine courte, l'anhydride glutarique (GA), contenant seulement 5 atomes de carbone, et un à longue chaine (600 atomes), le polyéthylène glycol (PEG) choisi aussi pour ses propriétés anti-adhérentes. Tout d'abord, cette stratégie a été développée sur des échantillons plats, qui facilitaient grandement les analyses de surface, telles que XPS et ToF-SIMS, et donc les processus d'optimisation de chaque étape, comme la résistance à la déformation, corrosion et l'analyse des propriétés biologiques. Ceci a permis de démontrer que le prétraitement de surface optimal pour les substrats L605 était l'électropolissage, agissant sur sa couche d'oxyde passive pour une efficacité maximale lors de l'étape d'amination. Le bras de liaison qui a démontré le plus grand potentiel pour immobiliser le peptide d'intérêt est le PEG, avec une augmentation significative de la migration et viabilité des cellules endothéliales, par rapport au substrat métallique nu. De plus, le greffage du peptide sur le PEG ajoutait des propriétés anti-thrombotique et anti-inflammatoire par rapport aux échantillons électropolis. Ce procédé a été ensuite adapté à des stents, dont la configuration 3D est très complexe. Après optimisation, les stents pegylées + peptides biomimétiques (Plasma-P8RI) ont été testés in vivo par implantation dans les coronaires de porc porcin pendant 7 jours et 28 jours et leur potentiel de ré-endothélialisation et anti-resténotique ont été évalués. Il a été constaté que la stratégie proposée dans ce projet de recherche favorisait la ré-endothélialisation après 7 jours par rapport au DES (Drug Eluting Stent) commercial, et limitait l'adhérence des leucocytes et des plaquettes lorsque comparé au BMS (Bare Metal Stent). Après 28 jours d'implantation le diamètre luminal des artères n'était pas réduit sur le stent Plasma-P8RI, ce qui signifie que ces stents modifiés ne présentaient pas de risque de resténose, contrairement aux BMS. Ce projet de recherche a permis de développer et de valider une stratégie prometteuse consistant à immobiliser directement des molécules bioactives sur des dispositifs cardiovasculaires en L605, alliage de chrome-cobalt. A notre connaissance, cette approche n'a jamais été rapportée dans la littérature à notre connaissance. Cette stratégie originale, dépourvue des limites associées à l'usage des polymères et basée sur un procédé plasma, présente des avantages évidents et ouvre la voie vers le développement de dispositifs cardiovasculaires innovants.Cardiovascular diseases represent the leading cause of death in the world. Among them is atherosclerosis that characterizes by the formation of a plaque on the arterial walls that narrows the lumen diameter. This atherosclerotic plaque disrupts the blood flow and can be complicated by thrombosis which can ultimately lead to myocardial infarction. Efficient revascularization is mandatory to treat this disease and a percutaneous coronary intervention (PCI) is performed complemented with the deployment of a stent. Stents are tiny wire mesh that reopens the artery, re-establishing the blood flow whilst supporting the artery avoiding its collapse. Nevertheless, complications after stent implantation exist and in-stent restenosis (ISR) is one of the major concerns. This complication is characterized by the reduction of the lumen diameter, similar to an atherosclerotic plaque, and it is associated to the wound caused on the endothelium by the stent implantation followed by the over-proliferation of smooth muscle cells. One of the first strategies to decrease ISR involved the manufacture of stents using different alloys such as stainless steel, nitinol and cobalt chromium alloys (L605). The latest alloy, L605, has generated significant interest because it allows the fabrication of thinner devices, which have decreased post-implantation clinical complications. Nonetheless, despite the decrease in ISR, when compared to other alloys, the integration of L605 bare metal stents in the host tissue is minimal or inexistent. Thus, enhanced biological properties, such as endothelialisation, low thrombosis activity and anti-inflammatory behaviour represent mandatory requirements for clinical applications. To confer these properties onto metallic devices, polymeric-based coatings, as an intermediate layer to further functionalize with bioactive molecules, are often deposited. Nonetheless, major techniques to deposit these polymeric coatings involve the use of wet-chemistry and do not ensure total resistance during the stent implantation procedure due to lack of cohesion and delamination of the polymeric layer. Thus, a novel approach that foregoes this previously mandatory coating step was developed in this research project. This novel approach involves the use of plasma-based techniques to create functional groups (reactive amine groups, -NH₂), directly onto the metallic surface without modifying the bulk properties, that can be used as anchor points for the further grafting of bioactive molecules of interest. Briefly, this novel approach can be divided in 3 blocks: a) Surface preparation, b) plasma functionalization and c) bioactive molecule grafting. Throughout this research project the optimization of these main blocks was performed aiming for the desired cardiovascular application. Concerning block a), surface preparation, electropolishing, thermal treatments and plasma immersion ion implantations were performed to obtain an oxide layer deformation and corrosion resistant whilst demonstrating the highest direct plasma amination efficiency, for block b). Finally, as regards block c), bioactive molecule grafting, two different linking arms were studied to assess their impact on conformation, and the biological performance of a bioactive peptide derived from the platelet endothelial cell adhesion molecule (PECAM-1 or CD31) due to its pro-endothelialization, anti-inflammatory and anti-thrombotic potential: Glutaric anhydride (GA), as a short chain spacer of 5 carbons, and polyethylene glycol (PEG), as a long chain spacer with antifouling properties. Initially, this strategy was developed on flat samples where using a combination of high-resolution surface characterizations techniques, such as XPS and ToF-SIMS, and corrosion, deformation and biological tests it was confirmed that the optimal surface pre-treatment for L605 was electropolishing, due to its passive oxide layer and that it further allowed to obtain the highest amination efficiency. Furthermore, the best linking arm to immobilize the peptide was PEG, which demonstrated a significantly increase on endothelial cell viability with a faster migration, when compared to the bare metallic substrate. Moreover, peptides immobilized by PEG demonstrated that endothelial cells attached to the surface presented an anti-thrombotic and anti-inflammatory phenotype, when compared to electropolished samples. Thus, this biomimetic surface was selected for an in vivo trial in porcine model to evaluate its potential re-endothelialization and anti-restenotic activity. It was found that by directly attaching a CD31 agonist onto the bare metal stent by this strategy improved re-endothelialization after 7 days when compared to commercial DES, with further, low adhesion of leukocytes and platelets when compared to BMS. Moreover, after 28 days of implantation, Plasma-P8RI did not present a significant decrease on the lumen diameter, which was not the case for BMS that presented in-stent restenosis after this period. Overall, this research project allowed the development and validation of a promising strategy to directly immobilize bioactive molecules onto L605 cobalt chromium cardiovascular devices, providing clear advantages of medical devices currently on the market. Furthermore, to the best of our knowledge, such plasma-based multi-step strategy has never been previously reported in literature

Macroscale abundance patterns of hydromedusae in the temperate Southwestern Atlantic (27º-56º S)

Gelatinous organisms are crucial components of marine ecosystems and some species imply social and economic consequences. However, certain geographic areas, such as the temperate Southwestern Atlantic (SWA, 27 - 56 S), remain understudied in terms of jellyfish ecological data. We analyzed 3,727 plankton samples collected along ~6.7 million km2 over a 31-year period (1983–2014) to determine the occurrence, abundance, and diversity patterns of hydromedusae in the SWA. Analyses were made at both community and species levels. Two abundance hot spots of hydromedusae were identified, where values up to 2,480 ind. m-3 were recorded between 2003 and 2014. Liriope tetraphylla and Obelia spp. were the main responsible for recurrent peaks. Diversity indexes were in the range of those published for temperate areas worldwide, and some coastal zones showed values that can be considered moderate to high for a temperate neritic region. The community analysis yielded 10 groups following previously determined biogeographic schemes throughout the study area. This work enhances the knowledge of hydromedusae in the SWA and provides essential information about the current global warming context and the gelatinous zooplankton data necessity.Fil: Dutto, María Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto Argentino de Oceanografía. Universidad Nacional del Sur. Instituto Argentino de Oceanografía; ArgentinaFil: Chazarreta, Carlo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto Argentino de Oceanografía. Universidad Nacional del Sur. Instituto Argentino de Oceanografía; ArgentinaFil: Rodriguez, Carolina Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Schiariti, Agustin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina. Instituto Nacional de Investigaciones y Desarrollo Pesquero; ArgentinaFil: Diaz Briz, Luciana Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Genzano, Gabriel Nestor. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentin

Modelo de evaluación formativa para la práctica pedagógica en instituciones educativas públicas de primaria de José Leonardo Ortiz

La presente investigación titulada Modelo de evaluación formativa para la práctica pedagógica en instituciones educativas públicas de primaria de José Leonardo Ortiz es de tipo descriptiva propositiva, tuvo como objetivo proponer un modelo de evaluación formativa para la mejora de la práctica pedagógica de los docentes en las instituciones educativas públicas de primaria de José Leonardo Ortiz. Cuya población estuvo conformada 451 docentes de aula de ambos sexos que laboran en las 20 instituciones públicas de educación primaria existentes en el distrito de José Leonardo Ortiz, la variable dependiente para conocer el estado en que se encuentra la práctica pedagógica, para la cual se elaboró un instrumento tipo cuestionario para tal fin el mismo que será validado a criterio de juicio de expertos la presente investigación se utilizó el procedimiento de la metodología cuantitativa basada en primer lugar en analizar la problemática de la investigación relacionada a la práctica pedagógica de los docentes de educación primaria del distrito de José Leonardo Ortiz

Aperfeiçoamentos em uma framework para análise de folgas em sistemas sócio-técnicos complexos : aplicação em um laboratório químico

Medidas para gerenciamento de saúde e segurança em laboratórios são de extrema importância em laboratórios químicos. As pessoas que realizam qualquer atividade em um ambiente de laboratório estão expostas a diversos perigos e, consequentemente, existe o risco de ocorrência de eventos adversos para a saúde e segurança. Este trabalho foi desenvolvido em um laboratório químico de uma universidade federal tem como principal objetivo o aperfeiçoamento de uma framework que permite a realização de uma análise sistemática qualitativa e quantitativa das folgas presentes em um sistema sócio-técnico complexo. Ferramentas da Engenharia de Resiliência foram utilizadas para estudar o laboratório, o qual foi considerado como um sistema sócio-técnico complexo. Uma das características de um sistema resiliente é a capacidade de lidar com a variabilidade, o que pode ser obtido por meio de recursos de folgas (slack) no sistema. O uso da framework permitiu obter dados importantes para a análise do sistema e sugestões de melhorias. Os aperfeiçoamentos propostos na framework mostraram-se eficazes, principalmente na quantificação das folgas e variabilidades, em função da utilização do método AHP (Analytical Hierarchy Process) para a análise de dados. O método AHP tornou possível substituir o uso de questionários para toda a equipe por uma avaliação direcionada a especialistas. Ao utilizar o AHP, os dados podem ser adquiridos com maior rapidez. Outro ganho obtido com o uso do método AHP foi a possibilidade de redução de uma etapa da framework, tornando-a mais concisa.Measures for health and safety management are of paramount importance in chemical laboratories. People who perform any activity in a laboratory environment are exposed to a variety of hazards and consequently there is a risk of adverse health and safety events. This work was developed in a chemical laboratory of a federal university, and has as main objective the improvement of a framework that allows the accomplishment of a systematic qualitative and quantitative analysis of the slack present in a complex socio-technical system. Tools of Resilience Engineering were used for studying a chemical laboratory, which was considered as a complex socio-technical system. One of the characteristics of a resilient system is the ability to deal with variability, which can be obtained through slack resources in the system. This work was developed in a chemical laboratory of a federal university and consists in the improvement of a framework that allows the accomplishment of a systematic qualitative and quantitative analysis of the slack present in the system. The use of the framework allowed to obtain data important for the analysis of the system and suggestions for improvements. The improvements proposed in the framework proved to be effective, especially in the quantification of slack and variability, as a function of the AHP (Analytical Hierarchy Process) method for data collection. The AHP method made it possible to replace the use of questionnaires for the entire team by an expert team assessment. By using AHP, data can be acquired more quickly. Another gain obtained with the use of the AHP method was the possibility of reducing one stage of the framework, making it more concise

Análisis empírico del papel de las competencias generales en el marco de los estudios superiores

En plena implantación de los nuevos planes de estudio de acuerdo al EEES, las universidades se enfrentan a un nuevo modelo educativo basado en competencias: competencias específicas y competencias generales. Las competencias específicas están asociadas a la adquisición y desarrollo de conocimientos de un área en particular, mientras que las competencias generales son transversales al plan de estudios y definen capacidades, habilidades y/o aptitudes que el alumno debe desarrollar para aplicarlas a lo largo de su carrera profesional. El objetivo de este trabajo es proporcionar una guía al docente sobre las posibles mejoras para tratar el mayor número de competencias generales satisfactoriamente. Concretamente, se ha analizado la manera en la que los docentes están promoviendo y desarrollando las competencias generales con el objetivo de detectar carencias, mejoras y necesidades. El análisis se ha realizado sobre el profesorado de la Titulación de Graduado en Ingeniería del Software de la Universidad Politécnica de Madrid.Peer Reviewe

Effect of the Fe Nanoparticles Generated by Pulsed Plasma in Liquid in the Catalyzed Ozone Removal of Phenolphthalein

We have synthesized, in this work, zero valent iron (ZVI) nanoparticles to improve the efficiency of degradation of phenolphthalein catalyzed by ozone in aqueous solution. The Fe nanoparticles were obtained using the pulsed plasma in liquid (PPL) method with water as the liquid medium. Such nanoparticles have a mean size of 12 nm and are composed of ~80% Fe0, while the rest are a mixture of Fe+2 and Fe+3 oxides. The degradation of phenolphthalein was carried on a glass reactor injecting a constant amount of ozone and introducing different concentrations of Fe nanoparticles to the system. When using pure ozone, the percentage of degradation of phenolphthalein measured by colorimetry after one hour of reaction was 84%. However, when Fe nanoparticles are used, such percentage can be as high as 98% in 50 minutes of reaction. Furthermore, the degradation rate constant was 0.0334 min−1 with only ozone and it can be as high as 0.0733 min−1 with Fe nanoparticles. Finally, the total mineralization of phenolphthalein was obtained by total organic carbon (TOC) determinations. It is shown that when using only ozone, we obtained a percentage of mineralization of 49% and 96% when using the highest concentration of Fe nanoparticles

Schneiderian Membrane Thickness and Clinical Implications for Sinus Augmentation: A Systematic Review and Metaâ Regression Analyses

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141205/1/jper0888-sup-0001.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/141205/2/jper0888.pd

Evolution, and functional analysis of Natural Resistance-Associated Macrophage proteins (NRAMPs) from Theobroma cacao and their role in cadmium accumulation

The presence of the toxic metal cadmium (Cd2+) in certain foodstuffs is recognised as a global problem, and there is increasing legislative pressure to reduce the content of Cd in food. The present study was conducted on cacao (Theobroma cacao), the source of chocolate, and one of the crops known to accumulate Cd in certain conditions. There are a range of possible genetic and agronomic methods being tested as a route to such reduction. As part of a gene-based approach, we focused on the Natural Resistance-Associated Macrophage Proteins (NRAMPS), a family of proton/metal transporter proteins that are evolutionarily conserved across all species from bacteria to humans. The plant NRAMP gene family are of particular importance as they are responsible for uptake of the nutritionally vital divalent cations Fe2+, Mn2+, Zn2+, as well as Cd2+. We identified the five NRAMP genes in cacao, sequenced these genes and studied their expression in various organs. We then confirmed the expression patterns in response to variation in nutrient cation availability and addition of Cd2+. Functional analysis by expression in yeast provided evidence that NRAMP5 encoded a protein capable of Cd2+ transport, and suggested this gene as a target for genetic selection/modification

Increase of Circulating Monocyte–Platelet Conjugates in Rheumatoid Arthritis Responders to IL-6 Blockage

Monocytes; Rheumatoid arthritis; TocilizumabMonocitos; Artritis reumatoide; TocilizumabMonòcits; Artritis reumatoide; TocilizumabPlatelets (PLT) bind to a significant percentage of circulating monocytes and this immunomodulatory interaction is increased in several inflammatory and autoimmune conditions. The therapeutic blockage of IL-6 with Tocilizumab (TCZ) alters PLT and the phenotype and function of monocytes in rheumatoid arthritis (RA). However, the relationship between monocyte–PLT conjugates (CD14+PLT+) and clinical and immunological variables and the regulation of this interaction by IL-6 blockage are still unknown. Here, we compared the presence of monocyte–PLT conjugates (CD14+PLT+) and membrane CD162 expression using flow cytometry, and, by ELISA, the markers of PLT activation (sCD62P and sCD40L) in healthy donors (HD) and patients with long-standing RA before TCZ (baseline). We found higher percentages and absolute counts of CD14+PLT+, and higher plasmatic levels of sCD62P and sCD40L but lower CD162 expression on monocytes from RA patients than those from HD. Additionally, the levels of CD14+PLT+ inversely correlated with inflammatory parameters. Interestingly, 95% of patients with lower percentages of CD14+PLT+ and only 63% of patients with higher percentages of CD14+PLT+ achieved a EULAR-defined response at four weeks (p = 0.036). After TCZ, the percentage of CD14+PLT+ increased in 92% of RA patients who achieved 12 w-remission (p < 0.001). Our results suggest that the binding of PLTs has a modulatory effect, accentuated by the increased binding of PLTs to monocytes in response to the therapeutic blockage of IL-6.This study is supported by Instituto de Salud Carlos III and Fondos FEDER (PI17/00072 and PI20/00184)