¿Qué opinan los estudiantes de ingeniería de nuestras formas de enseñar?

La motivación es un elemento fundamental en el aprendizaje, sin ella, difícilmente existirá el esfuerzo necesario para aprender. El objetivo de este trabajo es conocer, desde la perspectiva de los estudiantes de ingeniería, qué modos de enseñar les resultan motivadores para su aprendizaje y en qué medida este impacto motivacional puede estar afectado por sus diferencias individuales. Conocer la opinión de los alumnos es importante, ya que no es la situación de aprendizaje en sí misma la que resulta determinante, sino el significado que tenga para el estudiante. En este estudio participaron un total de 501 alumnos de ingeniería de la Universidad Politécnica de Madrid. Se les aplicó el cuestionario EMQ-B que permite medir el valor motivacional de las diferentes pautas docentes evaluadas. En general, los resultados del análisis descriptivo permiten clasificar las estrategias evaluadas en tres grupos. El primero incluye las estrategias consideradas como altamente motivadoras; el segundo, las claramente rechazadas y el tercero, las estrategias a las que los alumnos les atribuyen un cierto valor motivador, pero no muy alto. Este último grupo resulta relevante porque incluye algunas de las estrategias con las que más se viene trabajando en la actualidad –trabajo en grupo,realización de trabajos prácticos, etc. El análisis de varianza sugiere que el género puede estar modulando dichas valoraciones motivacionales

CHO/LY-B Cell Growth under Limiting Sphingolipid Supply: Correlation between Lipid Composition and Biophysical Properties of Sphingolipid-Restricted Cell Membranes

Sphingolipids (SL) are ubiquitous in mammalian cell membranes, yet there is little data on the behavior of cells under SL- restriction conditions. LY- B cells derive from a CHO linein whichserine palmitoyl transferase (SPT), thus de novo SL synthesis, is suppressed, while maintaining the capacity of taking up and metabolizing exogenous sphingoid bases from the culture medium. In this study, LY- B cells were adapted to grow in a fetal bovine serum (FBS)- deficient medium to avoid external uptake of li-pids. The lowest FBS concentration that allowed LY- B cell growth, though at a slow rate, under our conditions was 0.04%, that is, 250- fold less than the standard (10%) concentration. Cells grown under limiting SL concentrations remained viable for at least 72hours. Enriching with sphingomyelin the SL- deficient medium allowed the recovery of growth rates analogous to those of control LY- B cells. Studies in-cluding whole cells, plasma membrane preparations, and derived lipid vesicles were carried out. Laurdan fluorescence was recorded to measure membrane molecular order, showing a significant decrease in the rigidity of LY- B cells, not only in plasma membrane but also in whole cell lipid extract, as a result of SL limitation in the growth medium. Plasma membrane preparations and whole cell lipid extracts were also studied using atomic force microscopy in the force spectroscopy mode. Force measurements demonstrated that lower breakthrough forces were required to pen-etrate samples obtained from SL- poor LY- B cells than those obtained from control cells. Mass- spectroscopic analysis was also a helpful tool to understand the rear-rangement undergone by the LY- B cell lipid metabolism. The most abundant SL in LY- B cells, sphingomyelin, decreased by about 85% as a result of SL limitation in the medium, the bioactive lipid ceramide and the ganglioside precursor hexosylcera-mide decreased similarly, together with cholesterol. Quantitative SL analysis showed that a 250- fold reduction in sphingolipid supply to LY- B cells led only to a sixfold decrease in membrane sphingolipids, underlining the resistance to changes in com-position of these cells. Plasma membrane compositions exhibited similar changes, at least qualitatively, as the whole cells with SL restriction. A linear correlation was observed between the sphingomyelin concentration in the membranes, the degree of lipid order as measured by laurdan fluorescence, and membrane breakthrough forces assessed by atomic force microscopy. Smaller, though significant, changes were also detected in glycerophospholipids under SL- restriction conditions.This work was supported in part by grants from the Spanish Ministry of Economy (grant FEDER MINECO PGC2018-099857-B-I00) and the Basque Government (grants No. IT1264-19 and IT1270-19), as well as Fundación Biofísica Bizkaia and the Basque Excellence Research Centre (BERC) program of the Basque Government, and by the Swiss National Science Foundation (310030-184949

Patches and Blebs: A Comparative Study of the Composition and Biophysical Properties of Two Plasma Membrane Preparations from CHO Cells

This study was aimed at preparing and characterizing plasma membranes (PM) from Chinese Hamster Ovary (CHO) cells. Two methods of PM preparation were applied, one based on adhering cells to a poly-lysine-coated surface, followed by hypotonic lysis and removal of intracellular components, so that PM patches remain adhered to each other, and a second one consisting of bleb induction in cells, followed by separation of giant plasma membrane vesicles (GPMV). Both methods gave rise to PM in sufficient amounts to allow biophysical and biochemical characterization. Laurdan generalized polarization was used to measure molecular order in membranes, PM preparations were clearly more ordered than the average cell membranes (GP ≈0.450 vs. ≈0.20 respectively). Atomic force microscopy was used in the force spectroscopy mode to measure breakthrough forces of PM, both PM preparations provided values in the 4–6 nN range, while the corresponding value for whole cell lipid extracts was ≈2 nN. Lipidomic analysis of the PM preparations revealed that, as compared to the average cell membranes, PM were enriched in phospholipids containing 30–32 C atoms in their acyl chains but were relatively poor in those containing 34–40 C atoms. PM contained more saturated and less polyunsaturated fatty acids than the average cell membranes. Blebs (GPMV) and patches were very similar in their lipid composition, except that blebs contained four-fold the amount of cholesterol of patches (≈23 vs. ≈6 mol% total membrane lipids) while the average cell lipids contained 3 mol%. The differences in lipid composition are in agreement with the observed variations in physical properties between PM and whole cell membranes.This work was supported in part by grant PGC2018-099857-B-I00 (MCI/AEI/FEDER, UE) and grants IT1264-19 and IT1270-19 from the Basque Government

Plasma membrane effects of sphingolipid-synthesis inhibition by myriocin in CHO cells: a biophysical and lipidomic study

[EN] Suppression of a specific gene effect can be achieved by genetic as well as chemical methods. Each approach may hide unexpected drawbacks, usually in the form of side effects. In the present study, the specific inhibitor myriocin was used to block serine palmitoyltransferase (SPT), the first enzyme in the sphingolipid synthetic pathway, in CHO cells. The subsequent biophysical changes in plasma membranes were measured and compared with results obtained with a genetically modified CHO cell line containing a defective SPT (the LY-B cell line). Similar effects were observed with both approaches: sphingomyelin values were markedly decreased in myriocin-treated CHO cells and, in consequence, their membrane molecular order (measured as laurdan general polarization) and mechanical resistance (AFM-measured breakthrough force values) became lower than in the native, non-treated cells. Cells treated with myriocin reacted homeostatically to maintain membrane order, synthesizing more fully saturated and less polyunsaturated GPL than the non-treated ones, although they achieved it only partially, their plasma membranes remaining slightly more fluid and more penetrable than those from the control cells. The good agreement between results obtained with very different tools, such as genetically modified and chemically treated cells, reinforces the use of both methods and demonstrates that both are adequate for their intended use, i.e. the complete and specific inhibition of sphingolipid synthesis in CHO cells, without apparent unexpected effects.The authors are grateful to Dr. X. Contreras for his critical reading of the manuscript. This work was supported in part by the Spanish Ministerio de Ciencia e Innovacion (MCI), Agencia Estatal de Investigacion (AEI) and Fondo Europeo de Desarrollo Regional (FEDER) (grant No. PGC2018-099857-B-I00), by the Basque Government (Grants No. IT1264-19, IT1196-19 and IT1270-19), by the Fundacion Biofisica Bizkaia and by the Basque Excellence Research Centre (BERC) program of the Basque Government, and by the Swiss National Science Foundation (310030-184949)

The role of the interactome in the maintenance of deleterious variability in human populations

Recent genomic projects have revealed the existence of an unexpectedly large amount of deleterious variability in the human genome. Several hypotheses have been proposed to explain such an apparently high mutational load. However, the mechanisms by which deleterious mutations in some genes cause a pathological effect but are apparently innocuous in other genes remain largely unknown. This study searched for deleterious variants in the 1,000 genomes populations, as well as in a newly sequenced population of 252 healthy Spanish indi-viduals. In addition, variants causative of monogenic diseases and somatic variants from 41 chronic lymphocytic leukaemia patients were analysed. The deleterious variants found were analysed in the context of the interactome to understand the role of network topology in the maintenance of the observe

Sphingosine induces the aggregation of imine-containing peroxidized vesicles

AbstractLipid peroxidation plays a central role in the pathogenesis of many diseases like atherosclerosis and multiple sclerosis. We have analyzed the interaction of sphingosine with peroxidized bilayers in model membranes. Cu2+ induced peroxidation was checked following UV absorbance at 245nm, and also using the novel Avanti snoopers®. Mass spectrometry confirms the oxidation of phospholipid unsaturated chains. Our results show that sphingosine causes aggregation of Cu2+-peroxidized vesicles. We observed that aggregation is facilitated by the presence of negatively-charged phospholipids in the membrane, and inhibited by anti-oxidants e.g. BHT. Interestingly, long-chain alkylamines (C18, C16) but not their short-chain analogues (C10, C6, C1) can substitute sphingosine as promoters of vesicle aggregation. Furthermore, sphinganine but not sphingosine-1-phosphate can mimic this effect. Formation of imines in the membrane upon peroxidation was detected by 1H-NMR and it appeared to be necessary for the aggregation effect. 31P-NMR spectroscopy reveals that sphingosine facilitates formation of non-lamellar phase in parallel with vesicle aggregation. The data might suggest a role for sphingosine in the pathogenesis of atherosclerosis

Follow-up of late-onset Pompe disease patients with muscle magnetic resonance imaging reveals increase in fat replacement in skeletal muscles

Altres ajuts: This investigation was sponsored by the following grants, one from Sanofi Genzyme and another from the Spanish Ministry of Health, Fondos FEDER-ISCIII. Isabel Illa has received speaker honorarium from Grifols and Sanofi-Genzyme. Jordi Díaz-Manera has received speaker honorarium from PTC Therapeutics and Sanofi-Genzyme. The authors of this manuscript certify that they comply with the ethical guidelines for authorship and publishing in the Journal of Cachexia, Sarcopenia, and Muscle.42Late-onset Pompe disease (LOPD) is a genetic disorder characterized by progressive degeneration of the skeletal muscles produced by a deficiency of the enzyme acid alpha-glucosidase. Enzymatic replacement therapy with recombinant human alpha-glucosidase seems to reduce the progression of the disease; although at the moment, it is not completely clear to what extent. Quantitative muscle magnetic resonance imaging (qMRI) is a good biomarker for the follow-up of fat replacement in neuromuscular disorders. The aim of this study was to describe the changes observed in fat replacement in skeletal muscles using qMRI in a cohort of LOPD patients followed prospectively. A total of 36 LOPD patients were seen once every year for 4 years. qMRI, several muscle function tests, spirometry, activities of daily living scales, and quality-of-life scales were performed on each visit. Muscle MRI consisted of two-point Dixon studies of the trunk and thigh muscles. Computer analysis of the images provided the percentage of muscle degenerated and replaced by fat in every muscle (known as fat fraction). Longitudinal analysis of the measures was performed using linear mixed models applying the Greenhouse-Geisser test. We detected a statistically significant and continuous increase in mean thigh fat fraction both in treated (+5.8% in 3 years) and in pre-symptomatic patients (+2.6% in 3years) (Greenhouse-Geisser p < 0.05). As an average, fat fraction increased by 1.9% per year in treated patients, compared with 0.8% in pre-symptomatic patients. Fat fraction significantly increased in every muscle of the thighs. We observed a significant correlation between changes observed in fat fraction in qMRI and changes observed in the results of the muscle function tests performed. Moreover, we identified that muscle performance and mean thigh fat fraction at baseline visit were independent parameters influencing fat fraction progression over 4 years (analysis of covariance, p < 0.05). Our study identifies that skeletal muscle fat fraction continues to increase in patients with LOPD despite the treatment with enzymatic replacement therapy. These results suggest that the process of muscle degeneration is not stopped by the treatment and could impact muscle function over the years. Hereby, we show that fat fraction along with muscle function tests can be considered a good outcome measures for clinical trials in LOPD patients

Schoolbag weight carriage in Portuguese children and adolescents: a cross-sectional study comparing possible influencing factors

Background Schoolbags and the consequences of carrying them, particularly those associated with overload, are often studied as a health concern. Modifications in gait and posture were reported when children carried loads that corresponded to more than 10% of their body weight (BW). The aims of this study were to verify the load that is carried by Portuguese students and how it is influenced by factors such as school grade, school schedule, lunch site, physical education, sex and body mass index (BMI). Acquiring a more specific knowledge of the Portuguese context and understanding the influence of these factors may allow us to generate proposals to control them in ways that benefit students.MethodsThe load carried by students in the 5th grade (10.60.4years) and 9th grade (14.70.6years) were weighed with a luggage scale on all days of the week, resulting in 680 evaluations. Data related to the school day were also collected, such as the student's lunch site, how he or she got to school and his or her school schedule for that day. Individual height and weight were also assessed. Results The 5th grade students carried greater loads than the 9th grade students, resulting in a substantial difference relative to their BW. The school loads of the 5th grade students were mostly greater than 10% of their BWs. Girls tended to carry heavier loads than boys, and overweight students also tended to carry heavier loads. Students who could eat lunch at home carried less weight, and on physical education days, the total load carried increased, but the backpacks of the 5th grade students were lighter.Conclusions The results of the current study describe excessive schoolbag weight among Portuguese students and expound on some of the factors that influence it, which can help researchers and professionals design a solution to decrease children's schoolbag loads.Funds through FCT - Portuguese Foundation for Science and Technology (UID/DTP/04045/2019) - and the European Fund for regional development (FEDER) allocated by European Union through the COMPETE 2020 Programme (POCI-01-0145-FEDER-006969), and through the Project NanoSTIMA: Macro-to-Nano Human Sensing, Towards Integrated Multimodal Health Monitoring and Analytics (NORTE-01-0145-FEDER-000016), co-financed by Fundo Europeu de Desenvolvimento Regional (FEDER) -NORTE 2020

Quantitative muscle MRI to follow up late onset Pompe patients : a prospective study

Late onset Pompe disease (LOPD) is a slow, progressive disorder characterized by skeletal and respiratory muscle weakness. Enzyme replacement therapy (ERT) slows down the progression of muscle symptoms. Reliable biomarkers are needed to follow up ERT-treated and asymptomatic LOPD patients in clinical practice. In this study, 32 LOPD patients (22 symptomatic and 10 asymptomatic) underwent muscle MRI using 3-point Dixon and were evaluated at the time of the MRI with several motor function tests and patient-reported outcome measures, and again after one year. Muscle MRI showed a significant increase of 1.7% in the fat content of the thigh muscles in symptomatic LOPD patients. In contrast, there were no noteworthy differences between muscle function tests in the same period of time. We did not observe any significant changes either in muscle MRI or in muscle function tests in asymptomatic patients over the year. We conclude that 3-point Dixon muscle MRI is a useful tool for detecting changes in muscle structure in symptomatic LOPD patients and could become part of the current follow-up protocol in daily clinics

Thrombospondin-1 mediates muscle damage in brachio-cervical inflammatory myopathy and systemic sclerosis

To describe the clinical, serologic and histologic features of a cohort of patients with brachio-cervical inflammatory myopathy (BCIM) associated with systemic sclerosis (SSc) and unravel disease-specific pathophysiologic mechanisms occurring in these patients. We reviewed clinical, immunologic, muscle MRI, nailfold videocapillaroscopy, muscle biopsy, and response to treatment data from 8 patients with BCIM-SSc. We compared cytokine profiles between patients with BCIM-SSc and SSc without muscle involvement and controls. We analyzed the effect of the deregulated cytokines in vitro (fibroblasts, endothelial cells, and muscle cells) and in vivo. All patients with BCIM-SSc presented with muscle weakness involving cervical and proximal muscles of the upper limbs plus Raynaud syndrome, telangiectasia and/or sclerodactilia, hypotonia of the esophagus, and interstitial lung disease. Immunosuppressive treatment stopped the progression of the disease. Muscle biopsy showed pathologic changes including the presence of necrotic fibers, fibrosis, and reduced capillary number and size. Cytokines involved in inflammation, angiogenesis, and fibrosis were deregulated. Thrombospondin-1 (TSP-1), which participates in all these 3 processes, was upregulated in patients with BCIM-SSc. In vitro, TSP-1 and serum of patients with BCIM-SSc promoted proliferation and upregulation of collagen, fibronectin, and transforming growth factor beta in fibroblasts. TSP-1 disrupted vascular network, decreased muscle differentiation, and promoted hypotrophic myotubes. In vivo, TSP-1 increased fibrotic tissue and profibrotic macrophage infiltration in the muscle. Patients with SSc may present with a clinically and pathologically distinct myopathy. A prompt and correct diagnosis has important implications for treatment. Finally, TSP-1 may participate in the pathologic changes observed in muscle