Galectina-1: detección, caracterización, actividades biológicas e interacciones en plaquetas humanas

Las galectinas, previamente conocidas como lectinas de tipo S o S-Lac, son una familia filogenéticamente conservada que tienen homología de secuencias aminoacídicas y dominios de reconocimiento a carbohidratos (CRD). Todas las galectinas unen lactosa y otros oligosacáridos β-galactosídicos ejerciendo múltiples funciones. Se han identificado 15 galectinas de mamíferos, siendo designadas Gal-1 a la Gal-15. A través de estudios de localización de las galectinas se sabe que estas proteínas pueden secretarse en múltiples compartimentos celulares dependiendo del estatus celular. La Gal-1 pertenece a las galectinas prototipo, tiene un único CDR y puede formar homodímeros a través de interacciones no covalentes, lo que le confiere la habilidad de formar complejos multivalentes con glicoconjugados específicos en una amplia variedad de tejidos. La Gal-1 presenta una diversidad de comportamientos, presentando efectos estimulatorios o inhibitorios, de adhesividad o de anti-adhesividad, de proliferación o de inhibición de la proliferación, dependiendo del tipo celular, su estado de activación, expresión y su estado de glicosilación de receptores en su superficie, de la proporción monómero-dímero y de la distribución intra vs. extracelular. Las plaquetas son células sanguíneas anucleadas de forma discoide que circulan en una concentración de 150 a 450 x 109/L, formadas a partir de los megacariocitos como discos celulares pequeños que funcionan preservando la integridad del sistema vascular y sus acciones también están vinculadas con procesos complejos como inflamación y cáncer. Se sabe que aproximadamente el 15% del complejo GPIIb/IIIa plaquetario está compuesto por hidratos de carbono y que la interacción de la Gal-1 recombinante humana con plaquetas homólogas, provoca cambios conformacionales en los receptores GPIIb/IIIa los que favorecerían el fenómeno de activación. Debido a que la Gal-1 tiene un rol crucial en la homeostasis celular, inflamación y progresión tumoral, al igual que las plaquetas, y participa del funcionalismo plaquetario interaccionando a través de receptores/ligandos de la misma, nos propusimos purificar y caracterizar a la Gal-1 plaquetaria y plasmática humana, así como también establecer las interacciones con proteínas que participan en el funcionalismo de dichas células. En nuestro laboratorio, previamente demostramos que existía expresión de Gal-1endógena en plaquetas en reposo, tanto por IFI como por CF. Para el aislamiento de Gal-1 de las plaquetas y del plasma humano, realizamos cromatografía de intercambio iónico y cromatografía de afinidad en lactosa-agarosa. Encontramos que la Gal-1 plaquetaria co-purifica con actina, formando un complejo difícil de disociar. La presencia del complejo fue confirmada por western blot y por RP-HPLC-MS. Se estudió la actividad biológica de la Gal-1-actina mediante ensayos de hemoaglutinación utilizando glóbulos rojos de conejo tratados con neuraminidasa. A su vez, mediante microscopía confocal, se demostró que ambas proteínas co-localizaban tanto en plaquetas en reposo como en las activadas con Tr. Por otro lado, se realizaron estudios mediante IFI, para estudiar la interacción de Gal-1 con sus ligandos en plaquetas tanto en reposo como activadas con Tr y Gal-1, para dilucidar posibles interacciones con Ta y otras proteínas de la MEC. También purificamos la Gal-1 plasmática, pudiendo solo realizar la caracterización parcial de la misma. En base a nuestros resultados demostramos que la Gal- 1 se encuentra en muy baja concentración en plaquetas humanas, coincidentemente con los estudios de proteómica previos. Proponemos que la Gal-1 podría generar un complejo interactivo con la actina, la Ta y otras proteínas de la MEC en las plaquetas humanas.Facultad de Ciencias Exacta

Isolation of Galectin-1 from Human Platelets: Its Interaction with Actin

Galectins are a family of animal lectins defined by their β-galactoside-binding specificity and a consensus sequence in their carbohydrate-recognition domain. Galectin-1 (Gal-1) is expressed as a non-covalently linked homodimer present in a variety of tissues. Here we describe its isolation from human platelets by a procedure involving ionic exchange chromatography and affinity chromatography on lactose-agarose. Platelet Gal-1 co-purifies with actin, forming an actin-Gal-1 complex which does no dissociate even after treatment with sodium dodecyl sulfate. The presence of both proteins was confirmed by Western blot and by trypsin digestion followed by mass spectrometry identification. By hemagglutination assays we studied the response of recombinant Gal-1/actin, mixed and pre-incubated in different proportions, and then tested against neuraminidase treated rabbit red blood cells. The complex formation was confirmed by confocal microscopy, showing that both proteins co-localised in resting platelets as well as in thrombin-activated ones. These results suggest that endogenous Gal-1 forms an intracellular complex with monomeric actin and that, after platelet activation, Gal-1 could play a role in the polymerization-depolymerization process of actin, which concludes in platelet aggregation.Facultad de Ciencias Exacta

El diagnóstico ambiental en obras constructivas de cayo Cruz: premisa para implementar acciones de capacitación / The environmental diagnosis in construction cayo Cruz: premise to implement training actions

El objetivo del artículo es proponer un diagnóstico ambiental en zonas de desarrollo constructivo de cayo Cruz para la capacitación ambiental de los técnicos de la Asociación Económica Internacional Unión de Construcciones Militares Bouygues Bâtiment International en acciones preventivas de conservación. Para ello, se aplicó el diagnóstico en el área de intervención, se emplearon métodos como el analítico-sintético y el enfoque de sistema en la elaboración de las acciones y la estructuración de sus fundamentos teóricos. El análisis documental, la observación y el diagnóstico ambiental permitieron corroborar el estado actual y dinámica de los componentes del medio, así como las necesidades de capacitación ambiental de los técnicos de la AEI UCM– BBI. Se capacitaron 30 técnicos en la conservación de los componentes naturales y paisajísticos afectados por las parcelas de desarrollo turístico.The objective of the article is to propose an environmental diagnosis in areas of constructive development of Cruz Bay for the environmental training of technicians of the International Economic Association Union of Military Constructions Bouygues Bâtiment International in preventive conservation actions. To do this, the diagnosis was applied in the intervention area, using methods such as analytical-synthetic and system approach in the development of actions and the structuring of their theoretical foundations. The documentary analysis, observation and environmental diagnosis made it possible to corroborate the current and dynamic state of the environmental components, as well as the environmental training needs of the AEI UCM-BBI technicians. 30 technicians were trained in the conservation of the natural and landscape components affected by the plots of tourist development

Competence in Spiritual and Emotional Care: Learning Outcomes for the Evaluation of Nursing Students.

Spiritual and emotional care is an important part of the person, especially in situations such as changes in health or a community coping with a pandemic. However, nurses report scarce university training in this area of care. The aim of the study is to define a catalogue of learning outcomes for spiritual and emotional care for undergraduate nurses. The design used a mixed method for the development and validation of learning outcomes. The first phase designs the catalogue of learning outcomes through a coordinating group and uses a bibliographic search and nursing legislation. The second phase validates the proposal through a group of experts, with a questionnaire using the modified Delphi technique in two rounds. The initial proposal was 75 learning outcomes, of which 17 were eliminated, 36 changed their wording and the experts proposed 7 new ones. The experts validated 65 learning outcomes: 14 for Assessment and diagnosis; 5 for Planning; 17 for Intervention; 4 for Evaluation and quality; 8 for Communication and interpersonal relationship and 17 for Knowledge and intrapersonal development. In conclusion, the academic curriculum can include these learning outcomes to help undergraduate nurses in the process of acquiring knowledge, skills and attitudes in spiritual and emotional care.post-print641 K

Imbalance in the expression of genes associated with purinergic signalling in the lung and systemic arteries of COPDpatients.

Growing evidence indicates that purinergic signalling is involved in the pathogenesis of chronic obstructive pulmonary disease (COPD) and in the vascular remodelling that occurs in other disorders; however, its role in initial vascular changes of COPD is not entirely known. We hypothesised that expression of genes regulating extracellular ATP and adenosine levels would be altered in the lung and systemic arteries of COPD patients. Quantitative real-time PCR was performed to analyse the relative expression of 17 genes associated with purinergic signalling and inflammation in lungs and intercostal arteries of never smokers (NS) (n = 16), non-obstructed smokers (NOS) (n = 17) and COPD patients (n = 21). Gene expression of ATP-degrading enzymes was decreased in both tissues of NOS and COPD patients compared to NS. NT5E expression (gene transcribing for an AMP hydrolyzing ectonucleotidase) was increased in both tissues in NOS compared to the other groups. P1 and P2 receptors did not show changes in expression. Expression of genes associated with inflammation (interleukin-13) was upregulated only in lung tissues of COPD. These findings suggest that the expression of different extracellular ATP-degrading enzymes is altered in smokers (NOS and COPD patients), promoting inflammation. However, the high NT5E expression found only in NOS could compensate this inflammatory environment

The Use of Virtual Reality Facilitates Dialectical Behavior Therapy® “Observing Sounds and Visuals” Mindfulness Skills Training Exercises for a Latino Patient with Severe Burns: A Case Study

Sustaining a burn injury increases an individual’s risk of developing psychological problems such as generalized anxiety, negative emotions, depression, acute stress disorder, or post-traumatic stress disorder. Despite the growing use of Dialectical Behavioral Therapy® (DBT®) by clinical psychologists, to date, there are no published studies using standard DBT® or DBT® skills learning for severe burn patients. The current study explored the feasibility and clinical potential of using Immersive Virtual Reality (VR) enhanced DBT® mindfulness skills training to reduce negative emotions and increase positive emotions of a patient with severe burn injuries. The participant was a hospitalized (in house) 21-year-old Spanish speaking Latino male patient being treated for a large (>35% TBSA) severe flame burn injury. Methods: The patient looked into a pair of Oculus Rift DK2 virtual reality goggles to perceive the computer-generated virtual reality illusion of floating down a river, with rocks, boulders, trees, mountains, and clouds, while listening to DBT® mindfulness training audios during 4 VR sessions over a 1 month period. Study measures were administered before and after each VR session. Results: As predicted, the patient reported increased positive emotions and decreased negative emotions. The patient also accepted the VR mindfulness treatment technique. He reported the sessions helped him become more comfortable with his emotions and he wanted to keep using mindfulness after returning home. Conclusions: Dialectical Behavioral Therapy is an empirically validated treatment approach that has proved effective with non-burn patient populations for treating many of the psychological problems experienced by severe burn patients. The current case study explored for the first time, the use of immersive virtual reality enhanced DBT® mindfulness skills training with a burn patient. The patient reported reductions in negative emotions and increases in positive emotions, after VR DBT® mindfulness skills training. Immersive Virtual Reality is becoming widely available to mainstream consumers, and thus has the potential to make this treatment available to a much wider number of patient populations, including severe burn patients. Additional development, and controlled studies are needed

Increase in bloodstream infection due to vancomycin-susceptible enterococcus faecium in cancer patients: risk factors, molecular epidemiology and outcomes

We conducted a prospective study to assess the risk factors, molecular epidemiology and outcome of bloodstream infection (BSI) due to Enterococcus faecium in hospitalized cancer patients. Between 2006 and 2012, a significant increase in vancomycin-susceptible E. faecium BSI was observed among cancer patients. Comparison of 54 episodes of BSI due to E. faecium with 38 episodes of BSI due to E. faecalis showed that previous use of carbapenems was the only independent risk factor for E. faecium acquisition (OR 10.24; 95% CI, 1.35-77.66). All E. faecium isolates were susceptible to glycopeptides, whereas 97% showed high-level resistance to ampicillin and ciprofloxacin. All 30 isolates available for genotyping belonged to the hospital-associated E. faecium lineages 17, 18 and 78. After 2009, most of the isolates belonged to ST117 (lineage 78). Patients with E. faecium BSI were more likely to receive inadequate initial empirical antibiotic therapy than patients with E. faecalis BSI, and time to adequate empirical antibiotic therapy was also longer in the former group. No significant differences were found between the two groups regarding early and overall case-fatality rates. Independent risk factors for overall case-fatality were current corticosteroids (OR 4.18; 95% CI, 1.34-13.01) and intensive care unit admission (OR 9.97; 95% CI,1.96-50.63). The emergence of E. faecium among cancer patients is a concern since there are limited treatment options and it may presage the emergence of vancomycin-resistant enterococci. A rationale approach that combines infection control with antimicrobial stewardship

ECO D2.5 Learning analytics requirements and metrics report

In MOOCs, learning analytics have to be addressed to the various types of learners that participate. This deliverable describes indicators that enable both teachers and learner to monitor the progress and performance as well as identify whether there are learners at risk of dropping out. How these indicators should be computed and displayed to end users by means of dashboards is also explained. Furthermore a proposal based on xAPI statements for storing relevant data and events is provided.Part of the work carried out has been funded with support from the European Commission, under the ICT Policy Support Programme, as part of the Competitiveness and Innovation Framework Programme (CIP) in the ECO project under grant agreement n° 21127

Elearning, Communication and Open-data: Massive Mobile, Ubiquitous and Open Learning

ABSTRACT: In MOOCs, learning analytics have to be addressed to the various types of learners that participate. This deliverable describes indicators that enable both teachers and learner to monitor the progress and performance as well as identify whether there are learners at risk of dropping out. How these indicators should be computed and displayed to end users by means of dashboards is also explained. Furthermore a proposal based on xAPI statements for storing relevant data and events is provided