Identification of Proteases and Protease Inhibitors in Seeds of the Recalcitrant Forest Tree Species Quercus ilex.

Proteases and protease inhibitors have been identified in the recalcitrant species Quercus ilex using in silico and wet methods, with focus on those present in seeds during germination. In silico analyses showed that the Q. ilex transcriptome database contained 2,240 and 97 transcripts annotated as proteases and protease inhibitors, respectively. They belonged to the different families according to MEROPS, being the serine and metallo ones the most represented. The data were compared with those previously reported for other Quercus species, including Q. suber, Q. lobata, and Q. robur. Changes in proteases and protease inhibitors alongside seed germination in cotyledon and embryo axis tissues were assessed using proteomics and in vitro and in gel activity assays. Shotgun (LC-MSMS) analysis of embryo axes and cotyledons in nonviable (NV), mature (T1) and germinated (T3) seeds allowed the identification of 177 proteases and 12 protease inhibitors, mostly represented by serine and metallo types. Total protease activity, as determined by in vitro assays using azocasein as substrate, was higher in cotyledons than in embryo axes. There were not differences in activity among cotyledon samples, while embryo axis peaked at germinated T4 stage. Gel assays revealed the presence of protease activities in at least 10 resolved bands, in the Mr range of 60-260 kDa, being some of them common to cotyledons and embryo axes in either nonviable, mature, and germinated seeds. Bands showing quantitative or qualitative changes upon germination were observed in embryo axes but not in cotyledons at Mr values of 60-140 kDa. Proteomics shotgun analysis of the 10 bands with protease activity supported the results obtained in the overall proteome analysis, with 227 proteases and 3 protease inhibitors identified mostly represented by the serine, cysteine, and metallo families. The combined use of shotgun proteomics and protease activity measurements allowed the identification of tissue-specific (e.g., cysteine protease inhibitors in embryo axes of mature acorns) and stage-specific proteins (e.g., those associated with mobilization of storage proteins accumulated in T3 stage). Those proteins showing differences between nonviable and viable seeds could be related to viability, and those variables between mature and germinated could be associated with the germination process. These differences are observed mostly in embryo axes but not in cotyledons. Among them, those implicated in mobilization of reserve proteins, such as the cathepsin H cysteine protease and Clp proteases, and also the large number of subunits of the CNS and 26S proteasome complex differentially identified in embryos of the several stages suggests that protein degradation via CNS/26S plays a major role early in germination. Conversely, aspartic proteases such as nepenthesins were exclusively identified in NV seeds, so their presence could be used as indicator of nonviability.This research was funded by the Spanish Ministry of Economy and Competitiveness in the framework of Projects BIO2015- 64737-R and PID2019-109038RB-I00.S

Desarrollo de la técnica de FICTION como nueva herramienta para el diagnóstico precoz de cáncer de pulmón

El cáncer de pulmón es una de las causas de muerte más frecuentes en el mundo occidental. La supervivencia global de los pacientes no supera el 15% a los 5 años, debido principalmente a que la mayor parte de los casos se diagnostican en estadios avanzados. Además de la prevención primaria, mediante la reducción del consumo de tabaco, son necesarias nuevas tecnologías para el diagnóstico precoz de la enfermedad. Estudios recientes han demostrado que el TAC helicoidal del tórax es efectivo en la detección de nódulos pulmonares malignos en estadios precoces. En la actualidad se está valorando su eficacia en series amplias de pacientes de alto riesgo. Recientemente se ha desarrollado una nueva técnica de citogenética molecular, el FICTION (Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for the Investigation of Neoplasms). Esta técnica permite el análisis simultáneo de marcadores inmunofenotípicos y alteraciones genéticas presentes en las células tumorales. El objetivo de nuestro proyecto es su puesta a punto para el estudio de muestras de esputo y lavado broncoalveolar de pacientes con cáncer de pulmón. El fin último es estudiar la posibilidad de que esta técnica pueda ser utilizada, junto con el TAC helicoidal, en programas de detección precoz de cáncer de pulmón, para pacientes de alto riesgo. En este trabajo presentamos una revisión de la contribución de las distintas técnicas de citogenética al estudio del cáncer de pulmón y la metodología de trabajo que vamos a llevar a cabo en nuestro proyecto

The black hole of the transition process : dropout of care before transition age in adolescents

Recent evidence confirms the risks of discontinuity of care when young people make a transition from child and adolescent mental health services (CAMHS) to adult mental health services (AMHS), although robust data are still sparse. We aimed to identify when and how patients get lost to care during transition by tracking care pathways and identifying factors which influence dropping out of care during transition. This is a retrospective observational study of 760 patients who reached the transition age boundary within 12 months before transition time and being treated at CAMHS for at least during preceding 18 months. Data were collected at two time points: last visit to CAHMS and first visit to AHMS. Socio-demographic, clinical and service utilization variables on CAMHS treatment were collected. In the 12 months leading up to the transition boundary, 46.8% of subjects (n = 356) withdrew from CAHMS without further contact with AHMS, 9.3% withdrew from CAHMS but were referred to AHMS by other services, 29% were transferred from CAHMS to AHMS, 10% remained at CAHMS and 5% patients were transferred to alternative services. Fifty-six percent of subjects experience cessation of care before the transition age. The risk of dropout increases with shorter contact time in CAMHS, is greater in subjects without pharmacological treatment, and decreases in subjects with psychosis, bipolar disorder, eating disorders, mental retardation, and neurodevelopmental disorders. This study confirms that a large number of people drop out of care as they approach the CAMHS transition and experience discontinuity of care during this critical period

Allogeneic Hematopoietic Stem Cell Transplantation in Transformed Follicular Lymphoma (tFL): Results of a Retrospective Multicenter Study from GELTAMO/GETH-TC Spanish Groups

Simple Summary Follicular lymphoma (FL) is the most prevalent subtype of indolent lymphoma, accounting for 70% of all cases. The estimated risk of histological transformation (tFL), such as diffuse large B cell lymphoma (DLBCL), varies from 2-3% per year to 7-8% at 10 years in different series. Treatment after transformation is not clearly established. Allogeneic hematopoietic stem cell transplantation (alloSCT) could be an option for these patients, but it has not been widely explored. We analyze the efficacy and toxicity of alloSCT in 43 patients from 14 Spanish centers. We observed long-term survival in around one third of the patients, especially those who developed chronic graft versus host disease, indicating that alloSCT continues to be a potentially curative option for patients with tFL, mainly due to the graft versus lymphoma effect. Background: Transformation of follicular lymphoma into an aggressive lymphoma (tFL) worsens the prognosis and the standard treatment is not completely defined. Allogeneic hematopoietic stem cell transplantation (alloSCT) could be a potentially curative option for these patients, but it has not been widely explored. Methods: We designed a retrospective multicenter study to analyze the efficacy and toxicity of alloSCT in tFL patients and potential prognostic factors of survival. Results: A total of 43 patients diagnosed with tFL who underwent alloSCT in 14 Spanish centers between January 2000 and January 2019 were included. Median age was 44 (31-67) years. After a median follow-up of 58 months, estimated 5-year overall survival (OS) and progression-free survival (PFS) were both 35%. Estimated 100-day and 1-year non-relapse mortality (NRM) were 20% and 34%, respectively. The type of conditioning regimen (3-year OS of 52% vs. 20%, respectively, for reduced-intensity vs. myeloablative conditioning) and development of chronic graft versus host disease (cGVHD) (3-year OS of 75% vs. 40%) were the only factors significantly associated with OS. The only variable with an independent association with OS was cGVHD (HR, 3.4; 95% CI, 1.2-9.6). Conclusions: Our results indicate that alloSCT continues to be a potentially curative option for patients with tFL

Sociedad de la información e inclusión digital en España. Antecedentes. Primera parte

Se inicia con el cuestionamiento sobre el beneficio directo y tangible de las TIC en los países del "Sur", derivado de un modelo de desarrollo económico hecho en el "Norte", como elemento más constituyente de la brecha digital. El concepto brecha se aborda de acuerdo con los indicadores de la OCDE, por lo que se aportan datos estadísticos referidos a la proyección social de Internet y las potencialidades hacia la inclusión digital. Una de las medidas más eficaces en el esfuerzo inclusivo se considera la Educación y, particularmente, el e-learning, como su más eficaz instrumento para la alfabetización digital, . Las acciones necesitan modelos de actuación, por lo que se analizan las iniciativas europeas sobre la brecha digital, tanto en su ámbito interior (eEurope 2002 y 2005), como externo (CumbreMundial de la Sociedad de la información de Ginebra, 2003), así como también se registran las normas vigentes, basadas en desarrollo de experiencias, como el proyecto EL4EI, y orientadas hacia sectores en riesgo de exclusión, como es el caso de la mujer. El análisis del panorama europeo da paso al escenario español del desarrollo de la sociedad digital en sus antecedentes y en sus principales líneas, para llegar así al desarrollo del e-learning en España, cuyo concepto desde la óptica documentalista y sus beneficios"infonómicos" se destacan, para poder enfatizar sobre la implantación del e-learning en España en sectores como los contenidos, tecnologías y servicios, como también las tendencias del elearning, en el modelo de desarrollo económico y social español.The work begins with the issue about the direct and tangible benefit of the information and communication technologies in the so called countries of the “south ”, which is based on a economic development model elaborated in the “North”, as an element of the digital breach. Herein the concept breach is given according to de the indicators of the Organization for the Cooperation and Economical Development (OCDE in Spanish), for this reason are given statistical data about Internet’s social projection and potentialities for digital inclusion. Education, and particularly e-learning, is considered one of the most efficient steps towards digital inclusion, as the most efficient tool for digital literacy. Taking in account that actions need performance patterns, are analyzed European initiatives about the digital breach, internally (Europe 2002 and 2005) and externally (World Information Society Summit, held in Geneva, 2003), also are registered the existing standards, based in experiences, such as EL4EI, and oriented to sectors in risk of exclusion, such as women. After the analysis of the European countries, is reached the Spanish scenario regarding the development of the digital society and then reach the e-learning development in Spain, here are highlighted its “infonomic” benefits, so it could be possible emphasize e-learning implementation in Spain in areas like the contents, technologies and services, also are given the e-learning trends for the economic and social Spanish development pattern

Evolución histórica da implantación dos videoxogos na sociedade ourensá dende os anos oitenta ata a actualidade

Os últimos cincuenta anos veñen marcados por cambios importantes no ocio dos e das adolescentes, definidos fundamentalmente pola incorporación da tecnoloxía nas súas prácticas, e en particular pola introdución dos videoxogos. Estímase que en España o 80 % dos nenos/as de entre 6 e 10 anos e o 78 % de entre 11 e 14 anos xogan a videoxogos habitualmente (Asociación Española do Videoxogo –AEVI–, 2019). O obxectivo desta investigación foi identificar e describir os cambios nos patróns (tempo e actividades) do uso do tempo libre e do lecer dos e das adolescentes de 12-18 anos ourensáns, dende a introdución dos videoxogos a partir dos anos oitenta ata a actualidade. Para dar resposta formulouse un estudo dende un enfoque metodolóxico cualitativo, recorrendo á entrevista como principal instrumento de recollida de datos. Os resultados evidencian un incremento na cantidade de tempo dedicada aos videoxogos, que pasou de ser unha actividade complementaria a outras, a estar moi presente, así como un cambio no tipo de videoxogos empregados, que pasan de ser xogos máis sociais, onde se xoga en persoa, a xogo en liña. Ponse en valor algunhas das virtualidades do uso dos videoxogos (desenvemento de habilidades cognitivas e motoras), aínda que sobre todo destacan os perigos asociados: adicción, illamento social, competitividade e consumismo. Finalmente, tamén se aprecia unha marcada división sexo/xenérica en relación tanto coa práctica coma coa representación que mostran os videoxogos. Estes resultados son de gran relevancia para implementar políticas socioeducativas

Addressing challenges and opportunities of the European seafood sector under a circular economy framework

The European seafood and aquaculture sectors are facing important challenges in terms of environmental threats (climate change, marine debris, resources depletion), social development (worker rights, consumer's awareness) or economic growth (market and nonmarket goods and services, global competitiveness). These issues are forcing all stakeholders, from policy-makers to citizens and industries, to move to more sustainable policies, practices and processes. Consequently, an improvement in collaborations among different parties and beyond borders is required to create more efficient networks along the supply chain of seafood and aquaculture sectors. To achieve this, a "nexus thinking" approach (i.e. the analysis of actions in connected systems) combined with a life cycle thinking appears as an excellent opportunity to facilitate the transition to a circular economy.This work was supported by the EAPA_576/2018 NEPTUNUS project. The authors would like to acknowledge the financial support of Interreg Atlantic Area. A.C. Dias and P. Quinteiro acknowledge FCT/MCTES for the contracts CEECIND/02174/2017 and CEECIND/00143/2017, respectively, and for the financial support to CESAM (UID/AMB/50017/2019), through national funds. The Portuguese Foundation for Science and Technology (FCT) for supporting the contract of A. Marques in the framework of the IF2014 program (IF/00253/2014)

Lipoprotein(A) Concentrations In Rheumatoid Arthritis On Biologic Therapy: Results From The Cardiovascular In Rheumatology [Carma] Study Project

Background Plasma concentrations of lipoprotein (a) (Lp(a)), a lipoprotein with atherogenic and thrombogenic properties, have a strong genetic basis, although high concentrations of Lp(a) have also been reported in the context of inflammation, as in rheumatoid arthritis (RA). Few studies evaluate the impact of biologic therapies (BT) on Lp(a) in RA, taking into account that with these new therapies a better control of inflammation is achieved. Objective The aim of the study was to evaluate the plasma concentrations of Lp(a) in Spanish RA patients on BT attending rheumatology outpatient clinics. Methods Baseline analysis of the CARdiovascular in rheuMAtology project, a 10-year prospective study, evaluating the risk of cardiovascular events in RA and other forms of inflammatory arthritis. RA patients were classified according to treatment: no biologic, anti-tumor necrosis factor, anti-interleukin-6 receptor tocilizumab (TCZ), and other biologic (rituximab or abatacept). A model of linear multivariate regression was built in which the dependent variable was Lp(a) concentration and the explanatory variable was BT. The model was adjusted for confounding factors. Results Seven hundred and seventy-five RA patients were analyzed. Plasma concentrations of total cholesterol and triglyceride were significantly higher in TCZ-treated patients. Nevertheless, no significant difference in the atherogenic index between TCZ-treated patients and patients without BT was found. After adjusting for confounding factors, patients with BT had lower concentrations of Lp(a) than those without BT; however, only TCZ-treated patients achieved statistically significant differences (?: ?0.303, 95% confidence interval: ?0.558 to ?0.047; P = .02). Conclusions RA patients treated with TCZ show lower plasma concentrations of Lp(a) compared with patients without BT.This project has been supported by an unrestricted grant from Abbvie, Spain. The design, analysis, interpretation of results, and preparation of the article have been done independently of Abbvie. Dr González-Gay's studies have been supported by grants from “Fondo de Investigaciones Sanitarias” PI06/0024, PS09/00748, and PI12/00060 and RD12/0009/0013 (RIER) from “Instituto de Salud Carlos III” (ISCIII) (Spain)

Subtelomeric assembly of a multi-gene pathway for antimicrobial defense compounds in cereals

Non-random gene organization in eukaryotes plays a significant role in genome evolution. Here, we investigate the origin of a biosynthetic gene cluster for production of defence compounds in oat—the avenacin cluster. We elucidate the structure and organisation of this 12-gene cluster, characterise the last two missing pathway steps, and reconstitute the entire pathway in tobacco by transient expression. We show that the cluster has formed de novo since the divergence of oats in a subtelomeric region of the genome that lacks homology with other grasses, and that gene order is approximately colinear with the biosynthetic pathway. We speculate that the positioning of the late pathway genes furthest away from the telomere may mitigate against a ‘self-poisoning’ scenario in which toxic intermediates accumulate as a result of telomeric gene deletions. Our investigations reveal a striking example of adaptive evolution underpinned by remarkable genome plasticity

Long-Term Real-World Effectiveness and Safety of Ustekinumab in Crohn’s Disease Patients: The SUSTAIN Study

Background Large real-world-evidence studies are required to confirm the durability of response, effectiveness, and safety of ustekinumab in Crohn’s disease (CD) patients in real-world clinical practice. Methods A retrospective, multicentre study was conducted in Spain in patients with active CD who had received ≥1 intravenous dose of ustekinumab for ≥6 months. Primary outcome was ustekinumab retention rate; secondary outcomes were to identify predictive factors for drug retention, short-term remission (week 16), loss of response and predictive factors for short-term efficacy and loss of response, and ustekinumab safety. Results A total of 463 patients were included. Mean baseline Harvey-Bradshaw Index was 8.4. A total of 447 (96.5%) patients had received prior biologic therapy, 141 (30.5%) of whom had received ≥3 agents. In addition, 35.2% received concomitant immunosuppressants, and 47.1% had ≥1 abdominal surgery. At week 16, 56% had remission, 70% had response, and 26.1% required dose escalation or intensification; of these, 24.8% did not subsequently reduce dose. After a median follow-up of 15 months, 356 (77%) patients continued treatment. The incidence rate of ustekinumab discontinuation was 18% per patient-year of follow-up. Previous intestinal surgery and concomitant steroid treatment were associated with higher risk of ustekinumab discontinuation, while a maintenance schedule every 12 weeks had a lower risk; neither concomitant immunosuppressants nor the number of previous biologics were associated with ustekinumab discontinuation risk. Fifty adverse events were reported in 39 (8.4%) patients; 4 of them were severe (2 infections, 1 malignancy, and 1 fever). Conclusions Ustekinumab is effective and safe as short- and long-term treatment in a refractory cohort of CD patients in real-world clinical practice