Isolation of a laccase-coding gene from the lignin-degrading fungus Phlebia brevispora BAFC 633 and heterologous expression in Pichia pastoris

Aims: Isolate and characterize a laccase-encoding gene (lac I) of Phlebia brevispora BAFC 633, as well as cloning and expressing cDNA of lac I in Pichia pastoris. And to obtain a purified and characterized recombinant laccase to analyse the biotechnological application potential. Methods and Results: Lac I was cloned and sequenced, it contains 2447 pb obtained by PCR and long-distance inverse PCR. Upstream of the structural region of the laccase gene, response elements such as metals, antioxidants, copper, nitrogen and heat shock were found. The coding region consisted of a 1563-pb ORF encoding 521 amino acids. Lac I was functionally expressed in P. pastoris and it was shown that the gene cloned using the α-factor signal peptide was more efficient than the native signal sequence, in directing the secretion of the recombinant protein. Km and highest kcat/Km values towards ABTS, followed by 2,6-dimethylphenol, were similar to other laccases. Lac I showed tolerance to NaCl and solvents, and nine synthetic dyes could be degraded to different degrees. Conclusions: Lac I-encoding gene could be successfully sequenced having cis-acting elements located at the regulatory region. It was found that lac I cDNA expressed in P. pastoris using the α-factor signal peptide was more efficient than the native signal sequence. The purified Lac I exhibited high tolerance towards NaCl and various solvents and degraded some recalcitrant synthetic dyes. Significance and Impact of the Study: The cis-acting elements may be involved in the transcriptional regulation of laccase gene expression. These results may provide a further insight into potential ways of optimizing fermentation process and also open new frontiers for engineering strong promoters for laccase production. The Lac I stability in chloride and solvents and broad decolorization of synthetic dyes are important for its use in organic synthesis work and degradation of dyes from textile effluents respectively.Fil: Fonseca, Maria Isabel. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Molina, Melisa Antonella. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Winnik, D. L.. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; ArgentinaFil: Busi, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Fariña, Julia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Villalba, Laura. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Zapata, Pedro Dario. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Iron overload causes endolysosomal deficits modulated by NAADP-regulated two pore channels and RAB7A

Various neurodegenerative disorders are associated with increased brain iron content. Iron is known to cause oxidative stress, which concomitantly promotes cell death. Whereas endolysosomes are known to serve as intracellular iron storage organelles, the consequences of increased iron on endolysosomal functioning, and effects on cell viability upon modulation of endolysosomal iron release remain largely unknown. Here, we show that increasing intracellular iron causes endolysosomal alterations associated with impaired autophagic clearance of intracellular protein aggregates, increased cytosolic oxidative stress and increased cell death. These effects are subject to regulation by NAADP, a potent second messenger reported to target endolysosomal TPCNs (2-pore channels). Consistent with endolysosomal iron storage, cytosolic iron levels are modulated by NAADP, and increased cytosolic iron is detected when overexpressing active, but not inactive TPCNs, indicating that these channels can modulate endolysosomal iron release. Cell death triggered by altered intralysosomal iron handling is abrogated in the presence of an NAADP antagonist or when inhibiting RAB7A activity. Taken together, our results suggest that increased endolysosomal iron causes cell death associated with increased cytosolic oxidative stress as well as autophagic impairments, and these effects are subject to modulation by endolysosomal ion channel activity in a RAB7A-dependent manner. These data highlight alternative therapeutic strategies for neurodegenerative disorders associated with increased intracellular iron load

Metodología para el trabajo con géneros discursivos académicos

Este libro está destinado en primer lugar, a profesores, para que puedan aplicar las guías expuestas o elaborar otras similares con el mismo propósito; también a investigadores, en virtud de la «doble naturaleza» del discurso académico, por cuanto concierne a textos que responden a una situación comunicativa específica cuyos rasgos son, precisamente, el reflejo de los recursos expresivos que se pretenden enseñar mediante la práctica universitaria (Jiménez Calderón, 2015); y por último, a los estudiantes universitarios, que encontrarán en las guías expuestas una orientación muy específica para la elaboración de los géneros discursivos académicos a los que se refieren. INDICE: • Introducción: metodología para el trabajo con géneros discursivos académicos • Guía para la elaboración de abstracts • Guía para desenmarañar el comentario de textos lingüísticos • Pautas para el uso del aprendizaje colaborativo en una asignatura de lingüística • El comentario filológico de textos históricos • Elaboración de trabajos monográficos de primer curso • Guía para la elaboración de reseñas • Guía ortotipográfica para la presentación de trabajos de géneros académicos redactados en lengua francesa • Guía para la elaboración de trabajos académicos en el ámbito de la filología clásica • Guía para la elaboración de comentarios de textos poéticos en el área de literatura española • Guía para la elaboración de comentarios de textos narrativos en el área de literatura española • Guía para la elaboración de comentarios de textos literarios (dramática) • Guía para la elaboración de comentarios de textos literarios (ensayística) • El delicado asunto de las exposiciones orales… consejos de un neófito en la materia a un aprendiz de orador • Guía para la elaboración de comentarios de obras de historia del arte • Guía para la elaboración de trabajos monográficos en el área de historia del arte (nivel de grado) • Guía para la elaboración de trabajos prácticos en la fase inicial del grado en geografía y ordenación del territorio: «técnicas geográficas para el análisis territorial»This book is primarily intended for teachers, so that they can apply the guidelines or develop similar ones for the same purpose; Also to researchers, by virtue of the "double nature" of academic discourse, insofar as it concerns texts that respond to a specific communicative situation whose features are precisely the reflection of the expressive resources that are intended to be taught through university practice (Jiménez Calderón, 2015); And finally, to the university students, who will find in the guides exposed a very specific orientation for the elaboration of the academic discursive genres to which they refer

Towards a dynamic checklist of lichen-forming, lichenicolous and allied fungi of Ecuador – using the <i>Consortium of Lichen Herbaria</i> to manage fungal biodiversity in a megadiverse country

peer reviewedA checklist of Lichen-forming, Lichenicolous and Allied Fungi of Ecuador is presented with a total of 2599 species, of which 39 are reported for the first time from the country. The names of three species, Hypotrachyna montufariensis, H. subpartita and Sticta hypoglabra, previously not validly published, are validated. Pertusaria oahuensis, originally introduced by Magnusson as ‘ad interim’, is validated as Lepra oahuensis. The form Leucodermia leucomelos f. albociliata is validated. Two new combinations, Fissurina tectigera and F. timida, are made, and Physcia mobergii is introduced as a replacement name for the illegitimate P. lobulata Moberg non (Flörke) Arnold. In an initial step, the checklist was compiled by reviewing literature records of Ecuadorian lichen biota spanning from the late 19th century to the present day. Subsequently, records were added based on vouchers from 56 collections participating in the Consortium of Lichen Herbaria, a Symbiota-based biodiversity platform with particular focus on, but not exclusive to, North and South America. Symbiota provides sophisticated tools to manage biodiversity data, such as occurrence records, a taxonomic thesaurus, and checklists. The thesaurus keeps track of frequently changing names, distinguishing taxa currently accepted from ones considered synonyms. The software also provides tools to create and manage checklists, with an emphasis on selecting vouchers based on occurrence records that can be verified for identification accuracy. Advantages and limitations of creating checklists in Symbiota versus traditional ways of compiling these lists are discussed. Traditional checklists are well suited to document current knowledge as a ‘snapshot in time’. They are important baselines, frequently used by ecologists and conservation scientists as an established naming convention for citing species reported from a country. Compiling these lists, however, requires an immense effort, only to inadequately address the dynamic nature of scientific discovery. Traditional checklists are thus quickly out of date, particularly in groups with rapidly changing taxonomy, such as lichenized fungi. Especially in megadiverse countries, where new species and new occurrences continue to be discovered, traditional checklists are not easily updated; these lists necessarily fall short of efficiently managing immense data sets, and they rely primarily on secondary evidence (i.e. literature records rather than specimens). Ideally, best practices make use of dynamic database platforms such as Symbiota to assess occurrence records based both on literature citations and voucher specimens. Using modern data management tools comes with a learning curve. Systems like Symbiota are not necessarily intuitive and their functionality can still be improved, especially when handling literature records. However, online biodiversity data platforms have much potential in more efficiently managing and assessing large biodiversity data sets, particularly when investigating the lichen biota of megadiverse countries such as Ecuador

Effectiveness of an intervention for improving drug prescription in primary care patients with multimorbidity and polypharmacy:Study protocol of a cluster randomized clinical trial (Multi-PAP project)

This study was funded by the Fondo de Investigaciones Sanitarias ISCIII (Grant Numbers PI15/00276, PI15/00572, PI15/00996), REDISSEC (Project Numbers RD12/0001/0012, RD16/0001/0005), and the European Regional Development Fund ("A way to build Europe").Background: Multimorbidity is associated with negative effects both on people's health and on healthcare systems. A key problem linked to multimorbidity is polypharmacy, which in turn is associated with increased risk of partly preventable adverse effects, including mortality. The Ariadne principles describe a model of care based on a thorough assessment of diseases, treatments (and potential interactions), clinical status, context and preferences of patients with multimorbidity, with the aim of prioritizing and sharing realistic treatment goals that guide an individualized management. The aim of this study is to evaluate the effectiveness of a complex intervention that implements the Ariadne principles in a population of young-old patients with multimorbidity and polypharmacy. The intervention seeks to improve the appropriateness of prescribing in primary care (PC), as measured by the medication appropriateness index (MAI) score at 6 and 12months, as compared with usual care. Methods/Design: Design:pragmatic cluster randomized clinical trial. Unit of randomization: family physician (FP). Unit of analysis: patient. Scope: PC health centres in three autonomous communities: Aragon, Madrid, and Andalusia (Spain). Population: patients aged 65-74years with multimorbidity (≥3 chronic diseases) and polypharmacy (≥5 drugs prescribed in ≥3months). Sample size: n=400 (200 per study arm). Intervention: complex intervention based on the implementation of the Ariadne principles with two components: (1) FP training and (2) FP-patient interview. Outcomes: MAI score, health services use, quality of life (Euroqol 5D-5L), pharmacotherapy and adherence to treatment (Morisky-Green, Haynes-Sackett), and clinical and socio-demographic variables. Statistical analysis: primary outcome is the difference in MAI score between T0 and T1 and corresponding 95% confidence interval. Adjustment for confounding factors will be performed by multilevel analysis. All analyses will be carried out in accordance with the intention-to-treat principle. Discussion: It is essential to provide evidence concerning interventions on PC patients with polypharmacy and multimorbidity, conducted in the context of routine clinical practice, and involving young-old patients with significant potential for preventing negative health outcomes. Trial registration: Clinicaltrials.gov, NCT02866799Publisher PDFPeer reviewe

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio

5to. Congreso Internacional de Ciencia, Tecnología e Innovación para la Sociedad. Memoria académica

El V Congreso Internacional de Ciencia, Tecnología e Innovación para la Sociedad, CITIS 2019, realizado del 6 al 8 de febrero de 2019 y organizado por la Universidad Politécnica Salesiana, ofreció a la comunidad académica nacional e internacional una plataforma de comunicación unificada, dirigida a cubrir los problemas teóricos y prácticos de mayor impacto en la sociedad moderna desde la ingeniería. En esta edición, dedicada a los 25 años de vida de la UPS, los ejes temáticos estuvieron relacionados con la aplicación de la ciencia, el desarrollo tecnológico y la innovación en cinco pilares fundamentales de nuestra sociedad: la industria, la movilidad, la sostenibilidad ambiental, la información y las telecomunicaciones. El comité científico estuvo conformado formado por 48 investigadores procedentes de diez países: España, Reino Unido, Italia, Bélgica, México, Venezuela, Colombia, Brasil, Estados Unidos y Ecuador. Fueron recibidas un centenar de contribuciones, de las cuales 39 fueron aprobadas en forma de ponencias y 15 en formato poster. Estas contribuciones fueron presentadas de forma oral ante toda la comunidad académica que se dio cita en el Congreso, quienes desde el aula magna, el auditorio y la sala de usos múltiples de la Universidad Politécnica Salesiana, cumplieron respetuosamente la responsabilidad de representar a toda la sociedad en la revisión, aceptación y validación del conocimiento nuevo que fue presentado en cada exposición por los investigadores. Paralelo a las sesiones técnicas, el Congreso contó con espacios de presentación de posters científicos y cinco workshops en temáticas de vanguardia que cautivaron la atención de nuestros docentes y estudiantes. También en el marco del evento se impartieron un total de ocho conferencias magistrales en temas tan actuales como la gestión del conocimiento en la universidad-ecosistema, los retos y oportunidades de la industria 4.0, los avances de la investigación básica y aplicada en mecatrónica para el estudio de robots de nueva generación, la optimización en ingeniería con técnicas multi-objetivo, el desarrollo de las redes avanzadas en Latinoamérica y los mundos, la contaminación del aire debido al tránsito vehicular, el radón y los riesgos que representa este gas radiactivo para la salud humana, entre otros

Gestión del conocimiento. Perspectiva multidisciplinaria. Volumen 9

El libro “Gestión del Conocimiento. Perspectiva Multidisciplinaria”, volumen 9, de la Colección Unión Global, es resultado de investigaciones. Los capítulos del libro, son resultados de investigaciones desarrolladas por sus autores. El libro es una publicación internacional, seriada, continua, arbitrada de acceso abierto a todas las áreas del conocimiento, que cuenta con el esfuerzo de investigadores de varios países del mundo, orientada a contribuir con procesos de gestión del conocimiento científico, tecnológico y humanístico que consoliden la transformación del conocimiento en diferentes escenarios, tanto organizacionales como universitarios, para el desarrollo de habilidades cognitivas del quehacer diario. La gestión del conocimiento es un camino para consolidar una plataforma en las empresas públicas o privadas, entidades educativas, organizaciones no gubernamentales, ya sea generando políticas para todas las jerarquías o un modelo de gestión para la administración, donde es fundamental articular el conocimiento, los trabajadores, directivos, el espacio de trabajo, hacia la creación de ambientes propicios para el desarrollo integral de las instituciones