Um estudo sobre a função do objetivo instrucional na aprendizagem de competências contextuais

Neste estudo foi explorado o papel funcional que poderia ter o objetivo instrucional (OI) como evento de estímulo para os estudantes durante um episódio instrucional. O seu propósito foi estabelecer o efeito de apresentar objetivos instrucionais distintos para um mesmo discurso didático acerca da aprendizagem de competências contextuais de identificação. Participaram 26 estudantes de licenciatura distribuídos aleatoriamente em três grupos, dos quais duos receberam objetivos instrucionais distintos antes da leitura. O terceiro grupo não recebeu ninguém objetivo. Os resultados grupais nas tarefas de identificação não mostraram diferenças significativas entre os três grupos, mesmo se os melhores desempenhos de identificação corresponderam ao critério prescrito nos OI apresentados. Conclui-se que é necessário analisar a função de estímulo do OI em relação as interações com participantes individuais.This study examined the function of instructional objectives considered as stimuli events for students during instructional episodes. Its purpose was to explore the effect of presenting different instructional objectives for the same didactic discourse on the learning of contextual identification competences. Twenty six college students were distributed randomly in three groups, two of which received different pre-reading instructional objectives prescribing them to identify distinctive criteria in the same text. A third group did not receive any instructional objective. Results found in identification tasks did not show statistically significant differences between the three groups, even though the best performances were related to the criterion prescribed in the instructional objective. The authors conclude on the need to analyze the stimulus function of instructional objectives in terms of its interaction with individual participants.En este estudio se exploró el papel funcional que pudiera tener el objetivo instruccional (OI) como evento de estímulo para los estudiantes durante un episodio instruccional. Específicamente, su propósito fue determinar el efecto de presentar objetivos instruccionales diferentes para un mismo discurso didáctico sobre el aprendizaje de competencias contextuales de identificación. Participaron 26 estudiantes de licenciatura distribuidos al azar en tres grupos, dos de los cuales recibieron OI diferentes previos a la lectura, los que prescribían identificar distintos criterios en el mismo texto. El tercer grupo no recibió OI alguno. Los resultados grupales en las tareas de identificación no mostraron diferencias significativas entre los tres grupos, aunque los mejores desempeños de identificación correspondieron al criterio prescrito en los OI presentados. Se concluye en la necesidad de analizar la función de estímulo del OI en términos de las interacciones con participantes individuales

Copper effect on cytochrome b559 of photosystem II under photoinhibitory conditions

The definitive version is available at www.blackwell-synergy.comToxic Cu(II) effect on Cytochrome b559 under aerobic photoinhibitory conditions was examined in two different PSII membrane preparations active in oxygen evolution. The preparations differ in the content of Cytochrome b559 redox potential forms. Difference absorption spectra showed that the presence of Cu(II) induced the oxidation of the high-potential form of Cytochrome b559 in the dark. Addition of hydroquinone reduced the total oxidised high-potential form of Cytochrome b559 present in Cu(II)-treated PSII membranes indicating that no conversion to the low-potential form took place. Spectroscopic determinations of Cytochrome b559 during photoinhibitory treatment showed slower kinetics of Cu(II) effect on Cytochrome b559 as compared to the rapid loss of oxygen evolution activity in the same conditions. This result indicates that Cytochrome b559 is affected after PSII centers are photoinhibited. The high-potential form was more sensitive to toxic Cu(II) action than the low-potential form under illumination at pH 6.0. The content of the high-potential form of Cytochrome b559 was completely lost, however the low-potential content was unaffected in these conditions. This loss did not involve cytochrome protein degradation. Results are discussed in terms of different binding properties of the heme iron to the protonated or unprotonated histidine ligand in the high-potential and low-potential forms of Cytochrome b559, respectively.M. Bernal was recipient of an I3P Programme fellowship from Consejo Superior de Investigaciones Científicas. This work was supported by the Dirección General de Investigación (Grant BMC2002-00031) to R.P. and Gobierno de Aragón (Grant P015/2001) to I.Y., and it has been done within GC DGA 2002 Program of Gobierno de Aragón.Peer reviewe

El tráfico de cobre en el cloroplasto de plantas superiores: los transportadores de membrana

Introducción: El cobre (Cu) es un elemento esencial en la mayoría de organismos, aunque también puede ser tóxico a elevadas concentraciones (Festa y Thiele, 2011). En las plantas, el Cu participa en procesos fisiológicos como la fotosíntesis, la respiración mitocondrial, la percepción del etileno, el metabolismo de la pared celular, la protección frente al estrés oxidativo, la síntesis del cofactor de molibdeno y la regulación del ciclo circadiano (Puig y col., 2007; Mendel, 2013; Perea-García y col., 2010). Esto es debido a su habilidad para encontrarse en dos estados de oxidación diferentes in vivo, Cu+ y Cu2+. Por esta razón las plantas han desarrollado mecanismos para mantener la homeostasis, que permiten un eficiente uso del metal, a pesar del potencial daño que puede constituir para la célula (Ellingsen y col., 2007). En ella intervienen, transportadores de Cu de alta afinidad, metalochaperonas y ATPasas de tipo P (Ravet y Pilon, 2013). Muchos de los componentes de esta red han sido identificados y caracterizados durante los últimos años. Sin embargo, es necesaria más investigación en este campo para comprender completamente estos mecanismos. Por todo ello, el objetivo de esta Tesis Doctoral es el estudio de la homeostasis de Cu en plantas de soja (Glycine (G.) max), concretamente se ha realizado la caracterización de PAA2 (P-type ATPase in Arabidopsis 2), la ATPasa de tipo P1B encargada de transportar el Cu a través de la membrana tilaicoidal (Abdel-Ghany y col., 2005), y el estudio de la chaperona CCS (Copper chaperone for the Cu/Zn superoxide dismutase), encargada de la entrega del Cu a la Cu/Zn superóxido dismutasa (Pilon y col., 2011). Desarrollo: 1. Caracterización de PAA2. En la especie G. max, existen dos genes PAA2 (Bernal, 2006). Dichos genes, GmPAA2-1 y GmPAA2-2, tienen alta identidad entre ellos y se ha detectado un mecanismo de procesamiento alternativo en el gen GmPAA2-1, lo que podría conducir a la producción de una proteína completa (GmPAA2-1) y una proteína truncada (GmPAA2-1T). A su vez, el gen GmPAA2-2 retiene un hipotético intrón que produciría un codón de STOP prematuro y, por tanto, una proteína truncada. Utilizando la técnica de 5’ RACE, se vio que este gen tiene diferentes inicios de la transcripción según las condiciones nutricionales de Cu in vivo. Se ha realizado una caracterización de las proteínas GmPAA2-1 y GmPAA2-2 mediante la complementación funcional de los mutantes de S. cerevisiae ccc2, donde parecía que las proteínas GmPAA2-1, GmPAA2-1T o GmPAA2-2 no son capaces de complementar el fenotipo mutante. A su vez, en las levaduras silvestres, tampoco se observó un fenotipo apreciable por hipersensibilidad a cobre a consecuencia de la expresión de las proteínas de soja. Posteriormente se realizó la complementación del mutante paa1-1 de A. thaliana, que carece del transportador de Cu de la envuelta del cloroplasto (Shikanai y col., 2003), y se observó que las proteínas GmPAA2-1 y GmPAA2-2 no complementan el mutante paa1-1, indicando que las proteínas PAA1 y PAA2 tienen funciones que no se solapan. A continuación se realizó la complementación funcional del mutante paa2-1 de A. thaliana (Abdel-Ghany y col., 2005) con las proteínas GmPAA2-1 y GmPAA2-2. En este caso, GmPAA2-2 parece no complementar el fenotipo mutante paa2-1, en cambio, GmPAA2-1 es capaz de complementar el mutante paa2-1, lo que indica que la proteína de G. max es capaz de realizar la función de transporte de Cu+ a través de la membrana tilacoidal como en el caso de AtPAA2. Además, se ha estudiado la localización de GmPAA2-1 y GmPAA2-2 mediante expresión transitoria y estable de las proteínas fusionadas a YFP. Los datos indican que ambas proteínas tienen una localización cloroplástica. También se estudió la expresión de los genes mediante la técnica de qRT-PCR, y se vio que la expresión del gen GmPAA2-1 aumenta significativamente en algunas de las hojas según el aporte de Cu en la planta. En cambio, el gen GmPAA2-2 no parece mostrar una regulación por Cu. La localización in vivo de las diferentes proteínas GmPAA2 de soja se estudió mediante western blot con anticuerpos específicos. Los resultados mostraron bandas que podrían corresponder con las proteínas GmPAA2-1 y GmPAA2-1T, aunque no se observó la proteína GmPAA2-2. A pesar de ello, serán necesarios más estudios para conocer la función y caracterizar las nuevas posibles proteínas adicionales de soja. 2. Caracterización de la chaperona CCS de soja. La proteína GmCCS recombinante se sobreexpresó en E. coli y se purificó por cromatografía IMAC-Co2+ mayoritariamente en forma de apoproteína. Mediante estudios de interacción con su ligando por calorimetría isotérmica de titulación, se observó que la proteína GmCCS parece tener dos sitios de unión a Cu+ independientes con diferente afinidad y capacidad calorífica. La apoproteína GmCCS se encuentra en forma monomérica y dimérica, siendo más abundante la forma monomérica como indicaron los experimentos de cromatografía de exclusión molecular. La adición de Cu+ parece tener un efecto en el estado oligomérico, favoreciendo la dimerización de la proteína. Conclusiones: 1. La duplicación génica y los eventos de procesamiento alternativo pueden constituir un escenario favorable para la especiación, adaptación y evolución de las especies. En la especie G. max se han encontrado dos genes PAA2, con procesamiento alternativo en al menos uno de ellos, que podrían producir proteínas con funciones diferentes. Esto podría traducirse en una característica diferencial y, posiblemente, favorable para las plantas de esta especie. 2. Homólogos del gen PAA2 se han encontrado en todas las especies analizadas, lo que sugiere que su función es de gran importancia para las plantas. En soja, la proteína GmPAA2-1 parece realizar la función de transporte de Cu+ a través del tilacoide de manera similar a la proteína AtPAA2. A su vez, GmPAA2-1 tiene una función específica que no se superpone con la de PAA1 a pesar de compartir una gran similitud estructural con ésta. 3. La función de las proteínas adicionales GmPAA2 que podrían existir en la especie G. max es desconocida todavía. Es posible que realicen funciones alternativas al transporte de Cu+ a través de la membrana tilacoidal, pudiendo representar una característica positiva para las plantas que cuentan con ellas. Sin embargo, es necesaria más investigación para determinar estas cuestiones. 4. El mecanismo de actuación de la proteína CCS, así como su función de transferencia de Cu a su diana, Cu/ZnSOD, todavía cuentan con múltiples cuestiones por resolver. La proteína GmCCS parece encontrarse en dos estados oligoméricos diferentes (monómero y dímero), viéndose el dímero favorecido por la presencia de su ligando, Cu+. Esto podría tener una implicación en la interacción con la Cu/ZnSOD, mayoritariamente dimérica, en la que la transferencia del Cu podría producirse desde el sitio de unión a metal en el dominio III de GmCCS, con menor afinidad y mayor capacidad calorífica que el sitio de unión a metal en el dominio I

Transcriptome sequencing identifies SPL7-regulated copper acquisition genes FRO4/FRO5 and the copper dependence of iron homeostasis in Arabidopsis

24 Pags., 9 Figs., 2 Tabls., with Supplemental Data (15 Figs., 3 Tabls., 1 Method, 1 Data Set).The transition metal copper (Cu) is essential for all living organisms but is toxic when present in excess. To identify Cu deficiency responses comprehensively, we conducted genome-wide sequencing-based transcript profiling of Arabidopsis thaliana wild-type plants and of a mutant defective in the gene encoding SQUAMOSA PROMOTER BINDING PROTEIN-LIKE7 (SPL7), which acts as a transcriptional regulator of Cu deficiency responses. In response to Cu deficiency, FERRIC REDUCTASE OXIDASE5 (FRO5) and FRO4 transcript levels increased strongly, in an SPL7-dependent manner. Biochemical assays and confocal imaging of a Cu-specific fluorophore showed that high-affinity root Cu uptake requires prior FRO5/FRO4-dependent Cu(II)-specific reduction to Cu(I) and SPL7 function. Plant iron (Fe) deficiency markers were activated in Cu-deficient media, in which reduced growth of the spl7 mutant was partially rescued by Fe supplementation. Cultivation in Cu-deficient media caused a defect in root-to-shoot Fe translocation, which was exacerbated in spl7 and associated with a lack of ferroxidase activity. This is consistent with a possible role for a multicopper oxidase in Arabidopsis Fe homeostasis, as previously described in yeast, humans, and green algae. These insights into root Cu uptake and the interaction between Cu and Fe homeostasis will advance plant nutrition, crop breeding, and biogeochemical research.We acknowledge postdoctoral fellowships to M.B. from the Alexander von Humboldt Foundation and the Spanish Ministry of Science and Innovation; a Deutsche Forschungsgemeinshaft Heisenberg fellowship and funding from the FRONTIERS program at the University of Heidelberg, Germany, and the European Union InP Public Health Impact of Long-Term, Low-Level Mixed Element Exposure in Susceptible Population Strata (FOOD-CT-2006-016253) to U.K.; a grant from the National Science Foundation (IOS-0919739) to E.L.C.; a postdoctoral fellowship from the Spanish Foundation of Science and Technology (MEC-FECYT) to D.C.; National Institutes of Health Grant GM42143 to S.S.M.; and support from the University of California, Los Angeles–Department of Energy Institute for Genomics and Proteomics under Contract DE-FC02-02ER63421 to M.P.Peer reviewe

Preventive Conservation and Restoration Monitoring of Heritage Buildings Based on Fuzzy Logic

This article discusses the usability of the Art-Risk 3.0 software for research on the conservation of heritage buildings. It is a new and free software based on fuzzy logic, which enables the assessment of preventive conservation and surveillance of the restoration of heritage buildings over a period of time. This artificial intelligence-based tool considers the vulnerability of buildings, their environ ments, and their management to evaluate the necessity of their restoration or preventive con servation. To validate the Art-Risk 3.0, 500 theoretical case studies were analyzed, and a 14th century Mudejar-Gothic-style Church in Seville, Spain was studied both before and after its restora tion to identify post-restoration changes. This proof of concept demonstrates the capability of the Art-Risk 3.0 software to analyze environmental impacts on the vulnerability, risk, and functional service life of buildings, and assess the effectiveness of restoration activities. Additionally, this software identifies the most problematic factors and the necessity of restoration.Ministerio de Economía y Competitividad BIA2015- 64878-R (RETOS)Ministerio de Ciencia e Innovación PID2019-107257RB-I00 (FENIX)Ministerio de Ciencia, Innovación y Universidades EQC2019-005780-P (Ambulab-LAB)Junta de Andalucía PYC20 RE 034 UPO RESILIENT-TOURISMMinisterio de Ciencia e Innovación PTA2019-01688

Hydroxychloroquine is associated with a lower risk of polyautoimmunity: data from the RELESSER Registry

Objectives. This article estimates the frequency of polyautoimmunity and associated factors in a large retrospective cohort of patients with SLE. Methods. RELESSER (Spanish Society of Rheumatology Lupus Registry) is a nationwide multicentre, hospital-based registry of SLE patients. This is a cross-sectional study. The main variable was polyautoimmunity, which was defined as the co-occurrence of SLE and another autoimmune disease, such as autoimmune thyroiditis, RA, scleroderma, inflammatory myopathy and MCTD. We also recorded the presence of multiple autoimmune syndrome, secondary SS, secondary APS and a family history of autoimmune disease. Multiple logistic regression analysis was performed to investigate possible risk factors for polyautoimmunity. Results. Of the 3679 patients who fulfilled the criteria for SLE, 502 (13.6%) had polyautoimmunity. The most frequent types were autoimmune thyroiditis (7.9%), other systemic autoimmune diseases (6.2%), secondary SS (14.1%) and secondary APS (13.7%). Multiple autoimmune syndrome accounted for 10.2% of all cases of polyautoimmunity. A family history was recorded in 11.8%. According to the multivariate analysis, the factors associated with polyautoimmunity were female sex [odds ratio (95% CI), 1.72 (1.07, 2.72)], RP [1.63 (1.29, 2.05)], interstitial lung disease [3.35 (1.84, 6.01)], Jaccoud arthropathy [1.92 (1.40, 2.63)], anti-Ro/SSA and/or anti-La/SSB autoantibodies [2.03 (1.55, 2.67)], anti-RNP antibodies [1.48 (1.16, 1.90)], MTX [1.67 (1.26, 2.18)] and antimalarial drugs [0.50 (0.38, 0.67)]. Conclusion. Patients with SLE frequently present polyautoimmunity. We observed clinical and analytical characteristics associated with polyautoimmunity. Our finding that antimalarial drugs protected against polyautoimmunity should be verified in future studies

Cardiovascular events in Systemic Lupus Erythematosus: a nationwide study in Spain from the RELESSER Registry

This article estimates the frequency of cardiovascular (CV) events that occurred after diagnosis in a large Spanish cohort of patients with systemic lupus erythematosus (SLE) and investigates the main risk factors for atherosclerosis. RELESSER is a nationwide multicenter, hospital-based registry of SLE patients. This is a cross-sectional study. Demographic and clinical variables, the presence of traditional risk factors, and CV events were collected. A CV event was defined as a myocardial infarction, angina, stroke, and/or peripheral artery disease. Multiple logistic regression analysis was performed to investigate the possible risk factors for atherosclerosis. From 2011 to 2012, 3658 SLE patients were enrolled. Of these, 374 (10.9%) patients suffered at least a CV event. In 269 (7.4%) patients, the CV events occurred after SLE diagnosis (86.2% women, median [interquartile range] age 54.9 years [43.2-66.1], and SLE duration of 212.0 months [120.8-289.0]). Strokes (5.7%) were the most frequent CV event, followed by ischemic heart disease (3.8%) and peripheral artery disease (2.2%). Multivariate analysis identified age (odds ratio [95% confidence interval], 1.03 [1.02-1.04]), hypertension (1.71 [1.20-2.44]), smoking (1.48 [1.06-2.07]), diabetes (2.2 [1.32-3.74]), dyslipidemia (2.18 [1.54-3.09]), neurolupus (2.42 [1.56-3.75]), valvulopathy (2.44 [1.34-4.26]), serositis (1.54 [1.09-2.18]), antiphospholipid antibodies (1.57 [1.13-2.17]), low complement (1.81 [1.12-2.93]), and azathioprine (1.47 [1.04-2.07]) as risk factors for CV events. We have confirmed that SLE patients suffer a high prevalence of premature CV disease. Both traditional and nontraditional risk factors contribute to this higher prevalence. Although it needs to be verified with future studies, our study also shows-for the first time-an association between diabetes and CV events in SLE patients

Contribution of cardio-vascular risk factors to depressive status in the PREDIMED-PLUS Trial. A cross-sectional and a 2-year longitudinal study

Background Cardio-vascular disease and depression are thought to be closely related, due to shared risk factors. The aim of the study was to determine the association between cardio-vascular risk (CVR) factors and depressive status in a population (55-75 years) with metabolic syndrome (MetS) from the PREDIMED-Plus trial. Methods and findings Participants were classified into three groups of CVR according to the Framingham-based REGICOR function: (1) low (LR), (2) medium (MR) or (3) high/very high (HR). The Beck Depression Inventory-II (BDI-II) was used to assess depressive symptoms at baseline and after 2 years. The association between CVR and depressive status at baseline (n = 6545), and their changes after 2 years (n = 4566) were evaluated through multivariable regression models (logistic and linear models). HR women showed higher odds of depressive status than LR [OR (95% CI) = 1.78 (1.26, 2.50)]. MR and HR participants with total cholesterol <160 mg/mL showed higher odds of depression than LR [OR (95% CI) = 1.77 (1.13, 2.77) and 2.83 (1.25, 6.42) respectively)] but those with total cholesterol ¿280 mg/mL showed lower odds of depression than LR [OR (95% CI) = 0.26 (0.07, 0.98) and 0.23 (0.05, 0.95), respectively]. All participants decreased their BDI-II score after 2 years, being the decrease smaller in MR and HR diabetic compared to LR [adjusted mean±SE = -0.52±0.20, -0.41 ±0.27 and -1.25±0.31 respectively). MR and HR participants with total cholesterol between 240-279 mg/mL showed greater decreases in the BDI-II score compared to LR (adjusted mean±SE = -0.83±0.37, -0.77±0.64 and 0.97±0.52 respectively). Conclusions Improving cardiovascular health could prevent the onset of depression in the elderly. Diabetes and total cholesterol in individuals at high CVR, may play a specific role in the precise response.The PREDIMED-Plus trial was supported by the European Research Council through a grant to MAM (Advanced Research Grant 2013-2018; 340918). The project was also supported by the official funding agency for biomedical research of the Spanish Government (ISCIII) through the Fondo de Investigación para la Salud (FIS), which is co-funded by the European Regional Development Fund (four coordinated FIS projects), who awarded grants to JS and JV (PI13/00673, PI13/00492, PI13/00272, PI13/01123, PI13/00462, PI13/00233, PI13/02184, PI13/00728, PI13/01090, PI13/01056, PI14/01722, PI14/00636, PI14/00618, PI14/00696, PI14/01206, PI14/01919, PI14/00853, PI14/01374, PI16/00473, PI16/00662, PI16/01873, PI16/01094, PI16/00501, PI16/00533, PI16/00381, PI16/00366, PI16/01522, PI16/01120, PI17/00764, PI17/01183, PI17/00855, PI17/01347, PI17/00525, PI17/01827, PI17/00532, PI17/00215, PI17/01441, PI17/00508, PI17/01732 and PI17/00926). The International Nut&Dried Fruit Council-FESNAD also provided funding through a grant to MAM (201302), and Recercaixa also awarded a grant to JS (2013ACUP00194). The Department of Health, Generalitat de Cataluña by the calls 'Acció instrumental de programes de recerca orientats en lámbit de la recercaila innovació en salut' and 'Pla estrategic de recerca i innovació en salut (PERIS),' also awarded a grant to FF (SLT006/17/00246). This research was also partially funded by: Consejería de Salud de la Junta de Andalucía (PI0458/2013, PS0358/2016, PI0137/2018); Generalitat Valenciana (PROMETEO/2017/017); SEMERGEN, CIBEROBN, FEDER and ISCIII (CB06/03); EU-H2020 Grants (Eat2beNICE/h2020-sfs-2016-2, ref.728018; PRIME/h2020-SC1-BHC-2018-2020, ref: 847879)