Studies of the thioredoxin system in redox signaling and oxidative stress

Reactive oxygen species exert reversible posttranslational modifications in proteins containing redox sensitive thiols, thereby affecting several cellular processes and protein functions. Reductive enzyme systems, such as the thioredoxin system, can reverse most of these cysteine modifications. Therefore, a tight control between oxidizing and reducing events is the central principle of redox signaling. The studies within this thesis have covered several aspects of the Trx system in the modulation of cellular signaling pathways. In Paper I, we evaluated how b-AP15, a small inhibitor of proteasome-associated DUB, exerts redox perturbations in tumor cells. It is known that b-AP15triggers an increase level of reactive oxygen species (ROS) and proteotoxic stress in can- cer cells. However, its efficacy inducing apoptosis diminished by antioxidants. To identify the precise mechanism by which b-AP15 induces redox perturbations, we generated cells deprived of mitochondrial DNA. We found that in cells lacking mitochondria, the oxidative stress generated by b-AP15 was completely abrogated. Furthermore, to exclude that the observed increase in the levels of oxidative stress were due to an inhibition of TrxR1, we evaluated a number of proteasome asso- ciated DUBs inhibitors that did not inhibit TrxR1. Similarly, to b-AP15 all the inhibitors tested induced oxidative stress and the expression of HO-1. In parallel, we observed mitochondrial dysfunction, measuring the levels of COX5b and TOMM34, in both cases their respective levels decreased in those cells treated with b-AP15. Based in all the results we could conclude that the source of ROS in cells treated with b-AP15 was of mitochondrial origin. In Paper II, we performed a drug-screen of compounds sharing a common enone motif with b-AP15 and many natural products with antineoplastic effect. Through biochemical and structural analyses, we could demonstrate the binding of the enone containing compounds to the proteasome-associated cysteine deubiquitinase, USP14, inhibiting its activity. Additionally, we further analyzed a subset of those compounds in a zebrafish embryo model where they showed antineoplastic activity. These findings suggest that DUB inhibition is a relatively common mode of action by cytotoxic compounds containing motifs and it helps to explain the antineoplastic effects of natural products containing such functional group. In Paper III, we identified HRI as a redox-regulated protein, which becomes oxidized when activated upon As(III) exposure. TrxR1 associates with HRI in cells and together with TRP14 and Trx1 reduces HRI in vitro. Moreover, several specific inhibitors of TrxR1 lead to HRI-dependent eIF2α phosphorylation, trans- lation suppression and stress granule formation. Based on our finding that HRI- mediated translation suppression is essential for cellular survival under conditions of high As(III), we revealed the Trx system as a regulator of the HRI dependent translational stress response. In Paper IV, we evaluated the role of TRP14 in the regulation of different redox- regulated transcriptional factors using our unique tool pTRAF (plasmid for tran- scription factor reporter activation based upon fluorescence). We discovered that using TRP14 knockdown HEK293 cells, NRF2 activation increased upon treat- ment with auranofin, we also uncovered that TRP14 is crucial for HIFactivation upon TNF-α stimulation in hypoxic conditions. Furthermore, endogenous TRP14 levels increased under hypoxia or TNF-α treatment, suggesting that TRP14 could itself be regulated by NFκB and HIF, which is compatible with the presence of the corresponding response elements in the proximal TXNDC17 promoter region. Surprisingly, using TRP14 knockout HEK293 cells we found that global protein translation was reduced, which could be reverted with methionine or N- acetylcysteine supplementation. TRP14 knockout cells were also, in contrast to controls, highly sensitive to PPG. We conclude that TRP14 has several roles in control of redox signaling pathways, and that TRP14 is the main intracellular reductase for liberation of cysteine from cystine. In Paper V, we de novo synthesized eight vinyl sulfone compounds and evalu- ated their capacity to activate NRF2, NFκB and HIF1 in comparison with DMF using our previously mention tool pTRAF. We selected a set of compounds that activate NRF2 more selectively than DMF and characterize their downstream effects using in vitro and in vivo models. Our selected compounds display a more selective oligodendrocyte associated effect which could be explore in the future as a regenerative drug in demyelinating disorders

高中教师的工作对大学生个人责任和关系责任的预测

Several studies have shown the importance of training students in responsibility. Responsibility implies taking responsibility for one’s own actions (personal responsibility) and, assuming that our actions affect others (relational responsibility). Teachers can and should help train students in this dimension. The aim of this research is to analyze and understand which teaching behaviors and actions favor the formation of personal and relational responsibility in university students. The research design was sequential explanatory (mixed methodology). There were 403 university students for the quantitative study and 200 for the qualitative study. The measurement instrument used was designed and validated ad hoc. It presented adequate reliability and validity indicators. In addition, the ERRP scale was used, which assesses the personal and relational responsibility of the university student. The opinion of the participants was collected from open questions included in the questionnaire used. The results show that 14.4% of the students’ personal and relational responsibility can be explained by the teaching actions that contribute to it. The qualitative study shows that relational responsibility learning is higher than personal responsibility learning. A reliable and valid scale is provided to assess teachers’ actions that shape students’ personal and relational responsibility as well as teachers’ actions and behaviors that can help to elaborate a formative ethical competence on it.Diversos estudios han mostrado la importancia de formar a los estudiantes en responsabilidad lo que implica, hacerse cargo de las propias acciones (responsabilidad personal) y asumir que nuestras acciones afectan a los demás (responsabilidad relacional). Los docentes pueden y deben ayudar a formar a los estudiantes en esta dimensión. La investigación tiene como objetivo analizar y comprender qué comportamientos y acciones docentes favorecen la formación de la responsabilidad personal y relacional en los universitarios. El diseño de investigación fue secuencial explicativo (metodología mixta). Se contó con 403 estudiantes universitarios para el estudio cuantitativo y con 200 para el estudio cualitativo. El instrumento de medida empleado fue diseñado y validado ad hoc. Este presentó indicadores de fiabilidad y validez adecuados. Además, se empleó la escala ERRP que valora la responsabilidad personal y relacional del estudiante universitario. La opinión de los participantes fue recogida a partir de preguntas abiertas incluidas en el cuestionario utilizado. Los resultados muestran que el 14.4% de la responsabilidad personal y relacional de los estudiantes se puede explicar por las acciones docentes que contribuyen a la misma. El estudio cualitativo muestra que los aprendizajes de la responsabilidad relacional son mayores que los de la responsabilidad personal. Se aporta una escala fiable y válida que permite valorar las acciones de los docentes que forman la responsabilidad personal y relacional de los estudiantes así como acciones y comportamientos docentes que pueden ayudar a elaborar una competencia ética formativa sobre la misma.Ряд исследований показывает важность обучения учащихся ответственности, что подразумевает принятие ответственности за свои действия (личная ответственность) и принятие того, что их действия влияют на других (коллективная ответственность). Учителя могут и должны помочь в подготовке учащихся в этом аспекте. Цель исследования - проанализировать и понять, какое поведение и действия преподавателя способствуют формированию личной и коллективной ответственности у студентов университета. Дизайн исследования - последовательный разъяснительный (смешанная методология). В количественном исследовании приняли участие 403 студента университета, в качественном - 200. Используемый инструмент измерения был разработан и валидирован ad hoc. Он имел адекватные показатели надежности и валидности. Кроме того, использовалась шкала ERRP, оценивающая личную и коллективную ответственность студента университета. Мнение участников было собрано с помощью открытых вопросов, включенных в анкету. Результаты показали, что 14,4% личной и реляционной ответственности студентов можно объяснить действиями преподавателей, которые способствуют ее развитию. Качественное исследование показывает, что обучение коллективной ответственности выше, чем обучение личной ответственности. Предлагается надежная и валидная шкала для оценки действий преподавателей, формирующих личную и коллективую ответственность учащихся, а также действий и поведения преподавателей, которые могут помочь в развитии формируемой этической компетентности в этой области.各种研究表明培养学生责任感的重要性，这意味着对自己的行为负责（个人责任）并假设我们的行为会影响他人（关系责任）。教师可以而且应该在这个维度上帮助培养学生。该研究的目的是分析和了解哪些行为和教学行为有利于大学生个人和关系责任的形成。 研究采用解释性顺序（混合方法）设计。对 403 名大学生进行了定量研究，200 名大学生进行了定性研究。使用的测量仪器是专门设计和验证的。这提供了足够的可靠性和有效性指标。此外，还使用了 ERRP 量表，该量表评估大学生的个人和关系责任。参与者的意见是从所用问卷中包含的开放性问题中收集的。结果表明，学生的个人和关系责任中有 14.4% 可以用促成它的教学行为来解释。定性研究表明，关系责任的学习大于个人责任的学习。 本研究提供了一个可靠和有效的量表，允许评估教师的行为，这些行为促使学生的个人和关系责任的形成，以及有助于发展形成性道德能力的行为和教师行为

Gestión de datos de investigación en las universidades españolas: políticas y servicios

Ponencia presentada por el Grupo de Trabajo Repositorios de REBIUN, Acción 4, en el XVII Workshop REBIUN de Proyectos Digitales celebrado en la Universidad Pablo de Olavide (Sevilla) del 25 al 26 de octubre de 2018

Biodiesel production as a model of approach to natural sciences

Contextualizar los laboratorios de química, relacionándolos con situaciones de la vida diaria y contenidos disciplinares, motiva a despertar vocaciones científicas. El objetivo del trabajo fuefomentar el interés por las ciencias exactas y el cuidado medioambiental, a través de la participación activa de los estudiantes en la reutilización del aceite vegetal como modelo de laboratorio en contexto. Durante los talleres se estudió el origen de los aceites vegetales, las implicancias sobre la salud de la reiterada reutilización del aceite vegetal, las posibles formas de transformación en nuevos productos, la transesterificación, y la manipulación de reactivos químicos. Los destinatarios fueron estudiantes del nivel medio de 6 instituciones diferentes de laprovincia de Misiones (Argentina), ascendiendo a un total de 45 estudiantes de entre 17 a 21 años. Los talleres estuvieron a cargo de estudiantes, graduados y las directoras del proyecto. La metodología de la actividad consistió en una experiencia práctica y evaluación de la intervención. El empleo de un contexto socioambiental para la enseñanza de la química, generó concientización acerca de los cuidados del medioambiente; y nuevo conocimiento en relación con la química, al hacer uso de la reacción de transesterificación para la obtención de biodiesel.Contextualizing chemistry laboratories, relating them to daily life situations and disciplinary contents, motivates to awaken scientific vocations. The objective of the work was to promote interest in exact sciences and environmental care, through the active participation of students in the reuse of vegetable oil as a laboratory model in context. The origin of vegetable oils, the health implications of the repeated reuse of vegetable oil, the possible forms of transformation into new products, transesterification, and the manipulation of chemical reagents were studied during the workshops. Recipients were high school students from 6 different institutions in the province of Misiones (Argentina), reaching a total of 45 students between 17 and 21 years old. Workshops were led by students, graduates and the directors of the project. The methodology of the activity consisted of a practical experience and evaluation of the intervention. The use of a socioenvironmental context for the teaching of chemistry, generated awareness about environmental care; and new knowledge in relation to chemistry, by making use of the transesterification reaction to obtain biodiesel.Fil: Rodríguez, María Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Espinosa, Teresa Genara. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Gallará, Belén. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Oneto, María Eugenia. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Fernández Erbes, Loreley. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Krieger, Andrea Elizabeth. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Miguel, Nicolás. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Morel, Camila. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Zapata, Pedro Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; Argentin

Anti-IL-6 Receptor Tocilizumab in Refractory Graves? Orbitopathy: National Multicenter Observational Study of 48 Patients

Graves’ orbitopathy (GO) is the most common extrathyroidal manifestation of Graves’ disease (GD). Our aim was to assess the e cacy and safety of Tocilizumab (TCZ) in GO refractory to conventional therapy. This was an open-label multicenter study of glucocorticoid-resistant GO treated with TCZ. The main outcomes were the best-corrected visual acuity (BVCA), Clinical Activity Score (CAS) and intraocular pressure (IOP). These outcome variables were assessed at baseline, 1st, 3rd, 6th and 12th month after TCZ therapy onset. The severity of GO was assessed according to the European Group on Graves’ Orbitopathy (EUGOGO). We studied 48 (38 women and 10 men) patients (95 eyes); mean age standard deviation 51 11.8 years. Before TCZ and besides oral glucocorticoids, they had received IV methylprednisolone (n = 43), or selenium (n = 11). GO disease was moderate (n =29) or severe (n = 19) and dysthyroid optic neuropathy (DON) (n = 7). TCZ was used in monotherapy (n = 45) or combined (n = 3) at a dose of 8 mg/kg IV every four weeks (n = 43) or 162 mg/s.c. every week (n = 5). TCZ yielded a significant improvement in all of the main outcomes at the 1st month that was maintained at one year. Comparing the baseline with data at 1 year all of the variables improved; BCVA (0.78 0.25 vs. 0.9 0.16; p = 0.0001), CAS (4.64 1.5 vs. 1.05 1.27; p = 0.0001) and intraocular pressure (IOP) (19.05 4.1 vs. 16.73 3.4 mmHg; p = 0.007). After a mean follow-up of 16.1 2.1 months, low disease activity (CAS 3), was achieved in 88 eyes (92.6%) and TCZ was withdrawn in 29 cases due to low disease activity (n = 25) or ine cacy (n = 4). No serious adverse events were observed. In conclusion, TCZ is a useful and safe therapeutic option in refractory GO treatment.This work was also partially supported by RETICS Programs, RD08/0075 (RIER) and RD12/0009/0013 from “Instituto de Salud Carlos III” (ISCIII) (Spain)

Phenological Study of 53 Spanish Minority Grape Varieties to Search for Adaptation of Vitiviniculture to Climate Change Conditions

The main phenological stages (budburst, flowering, veraison, and ripeness) of 53 Spanish minority varieties were studied to determine their potential to help winegrowers adapt to climate change conditions. In total, 43 varieties were studied in the same location in Spain (Alcalá de Henares, in the Madrid region) and 10 varieties in 5 other regions (Galicia, Navarre, Catalonia, Extremadura, and Andalusia). Other traits of agronomic and oenological interest, such as yield and acidity, were also monitored. The results allow for the grouping of the varieties into several clusters according to the time of ripeness (very early—only for red varieties—and early, intermediate, and late, for both red and white varieties) and yield (high, medium, and low). The total acidity in the grape juice ranged from 3 to 11 g of tartaric acid/L. The average temperatures were higher (up to 3–4 °C during summer) compared to historical averages during the 1957–2021 time period. Advanced phenology phases and reduced acidity are regarded as negative effects of climate change for winegrowing practices. Since some minority varieties showed late or intermediate ripening, high acidity, and high (1 Kg/shoot) or medium (0.5 Kg/shoot) yield, our findings suggest that they may be cultivated in the coming years by winegrowers as an approach to mitigate climate change effects.Project RTI2018-101085-R-C31, “Valorization of Minority Grapevine Varieties for their Potential for Wine Diversification and Resilience to Climate Change (MINORVIN),” funded by MCIN/AEI/10.13039/501100011033, and by the ERDF, A Way to Make Europe.Peer reviewe

Cross-disease Meta-analysis of Genome-wide Association Studies for Systemic Sclerosis and Rheumatoid Arthritis Reveals IRF4 as a New Common Susceptibility Locus

Objectives: Systemic sclerosis (SSc) and rheumatoid arthritis (RA) are autoimmune diseases that share clinical and immunological characteristics. To date, several shared SSc- RA loci have been identified independently. In this study, we aimed to systematically search for new common SSc-RA loci through an inter-disease meta-GWAS strategy. Methods: We performed a meta-analysis combining GWAS datasets of SSc and RA using a strategy that allowed identification of loci with both same-direction and opposingdirection allelic effects. The top single-nucleotide polymorphisms (SNPs) were followed-up in independent SSc and RA case-control cohorts. This allowed us to increase the sample size to a total of 8,830 SSc patients, 16,870 RA patients and 43,393 controls. Results: The cross-disease meta-analysis of the GWAS datasets identified several loci with nominal association signals (P-value < 5 x 10-6), which also showed evidence of association in the disease-specific GWAS scan. These loci included several genomic regions not previously reported as shared loci, besides risk factors associated with both diseases in previous studies. The follow-up of the putatively new SSc-RA loci identified IRF4 as a shared risk factor for these two diseases (Pcombined = 3.29 x 10-12). In addition, the analysis of the biological relevance of the known SSc-RA shared loci pointed to the type I interferon and the interleukin 12 signaling pathways as the main common etiopathogenic factors. Conclusions: Our study has identified a novel shared locus, IRF4, for SSc and RA and highlighted the usefulness of cross-disease GWAS meta-analysis in the identification of common risk loci

Complement component C4 structural variation and quantitative traits contribute to sex-biased vulnerability in systemic sclerosis

Altres ajuts: Fondo Europeo de Desarrollo Regional (FEDER), "A way of making Europe".Copy number (CN) polymorphisms of complement C4 play distinct roles in many conditions, including immune-mediated diseases. We investigated the association of C4 CN with systemic sclerosis (SSc) risk. Imputed total C4, C4A, C4B, and HERV-K CN were analyzed in 26,633 individuals and validated in an independent cohort. Our results showed that higher C4 CN confers protection to SSc, and deviations from CN parity of C4A and C4B augmented risk. The protection contributed per copy of C4A and C4B differed by sex. Stronger protection was afforded by C4A in men and by C4B in women. C4 CN correlated well with its gene expression and serum protein levels, and less C4 was detected for both in SSc patients. Conditioned analysis suggests that C4 genetics strongly contributes to the SSc association within the major histocompatibility complex locus and highlights classical alleles and amino acid variants of HLA-DRB1 and HLA-DPB1 as C4-independent signals

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality