Generation of thermo-sensitive allele of the TPR like protein Nup211by PCR mutagenesis

Motivation: TPR proteins are conserved large coiled-coil proteins that localize at the nucleoplasmic side of the nuclear pore complex and participate in multiple aspects of DNA metabolism. The protein Nup211, fission yeast homolog of Mlp1/Mlp2/Tpr, participate in the mRNA export and is essential for vegetative growth. The aim of this work is to create a collection of thermo-sensitive alleles of nup211.Methods: To create the collection, we have generated a new strain with the nup211 gen tagged with GFP at the amino terminal extreme and confirmed by fluorescent microscopy that the protein Nup211 localized in the nuclear envelop. Then, we have carried out a Taq PCR-based Random Mutagenesis with reduced concentration of dATP. The PCR products were transformed into a wild type strain to generate conditional mutants. The transformants obtained whose growth was impaired at 36ºC were preselected as thermo-sensitive mutants. To confirm the growth deficiency of these clones, a drop assay was performed and the best candidates were selected. These thermo-sensitive mutants were cultivated at 25ºC as well as 36ºC and both cultures were subjected to various experiments in order to study any changes in the localization of Nup211.Results: Up to now, we have demonstrated by fluorescent microscopy that the thermo-sensitive mutants show a modified nuclear distribution of Nup211 and different cellular phenotype, suggesting that the differents clones might represent differents nup211 thermo-sensitive alleles. These alleles are going to be subjected to various experiments to clarify the role of the protein in the mRNA export

Bub1 as a recruitment platform for Spindle Assembly Checkpoint components

Motivation: The Spindle Assembly Checkpoint (SAC) is a safeguard mechanism conserved in all the eukaryotes that ensures the correct chromosome segregation in mitosis by preventing the premature mitotic exit in condition of unattached kinetochores. SAC defects lead to chromosome mis-segregation provoking aneuploidies that has been widely associated with cancer. Kinase Bub1 is a key player in SAC function because it maintains a proper centromeric cohesion and serves as a platform for other SAC components such as Mad1, Mad2 and Mad3. In this study, we are characterizing a bub1 mutant allele which exhibits an impaired SAC function. The phenotype shown by this mutant has not been previously described. This mutant can provide new insights about the Bub1-dependent recruitment of SAC components to the kinetochores and the mechanism of mitosis arrest.Methods: We made use of live imaging techniques to study SAC function by using GFP-tagged alleles of the main SAC components (like Mad1, Mad2 and Mad3), comparing the wild type background versus the bub1 mutant allele. Furthermore, we have used a cold sensitive tubulin mutant that allows us to test SAC activity; in response to microtubules damage induced by this mutant, cells activate the SAC and arrest in metaphase. We have also tested Bub1 protein levels in a wild-type and in the Bub1-mutant by Western Blot. Finally, we are performing a two-hybrid screening using a S. pombe library-strain to detect differences in the interactome of the Bub1-mutant compared with the wild-type. Results: We have demonstrated that in the Bub1-mutant background, Mad1 correctly localizes at the kinetochores meanwhile Mad2 does not. Bub1 protein levels turned out to be quite similar in both strain. Additionally, we have observed that the SAC defects notice in the Bub1-mutant in the cold sensitive background partially phenocopies the one seen in the Bub1-deleted cells.Conclusions: Our Bub1-mutant is unable to maintain a proper metaphase block in the cold-sensitive tubulin background and exhibit a SAC failure. We are working in a model where Bub1 could be regulating SAC activity by promoting Mad2 recruitment to kinetochores

Propagation of Policies in Rich Data Flows

Governing the life cycle of data on the web is a challenging issue for organisations and users. Data is distributed under certain policies that determine what actions are allowed and in which circumstances. Assessing what policies propagate to the output of a process is one crucial problem. Having a description of policies and data flow steps implies a huge number of propagation rules to be specified and computed (number of policies times number of actions). In this paper we provide a method to obtain an abstraction that allows to reduce the number of rules significantly. We use the Datanode ontology, a hierarchical organisation of the possible relations between data objects, to compact the knowledge base to a set of more abstract rules. After giving a definition of Policy Propagation Rule, we show (1) a methodology to abstract policy propagation rules based on an ontology, (2) how effective this methodology is when using the Datanode ontology, (3) how this ontology can evolve in order to better represent the behaviour of policy propagation rules

A922 Sequential measurement of 1 hour creatinine clearance (1-CRCL) in critically ill patients at risk of acute kidney injury (AKI)

Meeting abstrac

Differential clinical characteristics and prognosis of intraventricular conduction defects in patients with chronic heart failure

Intraventricular conduction defects (IVCDs) can impair prognosis of heart failure (HF), but their specific impact is not well established. This study aimed to analyse the clinical profile and outcomes of HF patients with LBBB, right bundle branch block (RBBB), left anterior fascicular block (LAFB), and no IVCDs. Clinical variables and outcomes after a median follow-up of 21 months were analysed in 1762 patients with chronic HF and LBBB (n = 532), RBBB (n = 134), LAFB (n = 154), and no IVCDs (n = 942). LBBB was associated with more marked LV dilation, depressed LVEF, and mitral valve regurgitation. Patients with RBBB presented overt signs of congestive HF and depressed right ventricular motion. The LAFB group presented intermediate clinical characteristics, and patients with no IVCDs were more often women with less enlarged left ventricles and less depressed LVEF. Death occurred in 332 patients (interannual mortality = 10.8%): cardiovascular in 257, extravascular in 61, and of unknown origin in 14 patients. Cardiac death occurred in 230 (pump failure in 171 and sudden death in 59). An adjusted Cox model showed higher risk of cardiac death and pump failure death in the LBBB and RBBB than in the LAFB and the no IVCD groups. LBBB and RBBB are associated with different clinical profiles and both are independent predictors of increased risk of cardiac death in patients with HF. A more favourable prognosis was observed in patients with LAFB and in those free of IVCDs. Further research in HF patients with RBBB is warranted

Evaluation of appendicitis risk prediction models in adults with suspected appendicitis

Background Appendicitis is the most common general surgical emergency worldwide, but its diagnosis remains challenging. The aim of this study was to determine whether existing risk prediction models can reliably identify patients presenting to hospital in the UK with acute right iliac fossa (RIF) pain who are at low risk of appendicitis. Methods A systematic search was completed to identify all existing appendicitis risk prediction models. Models were validated using UK data from an international prospective cohort study that captured consecutive patients aged 16–45 years presenting to hospital with acute RIF in March to June 2017. The main outcome was best achievable model specificity (proportion of patients who did not have appendicitis correctly classified as low risk) whilst maintaining a failure rate below 5 per cent (proportion of patients identified as low risk who actually had appendicitis). Results Some 5345 patients across 154 UK hospitals were identified, of which two‐thirds (3613 of 5345, 67·6 per cent) were women. Women were more than twice as likely to undergo surgery with removal of a histologically normal appendix (272 of 964, 28·2 per cent) than men (120 of 993, 12·1 per cent) (relative risk 2·33, 95 per cent c.i. 1·92 to 2·84; P < 0·001). Of 15 validated risk prediction models, the Adult Appendicitis Score performed best (cut‐off score 8 or less, specificity 63·1 per cent, failure rate 3·7 per cent). The Appendicitis Inflammatory Response Score performed best for men (cut‐off score 2 or less, specificity 24·7 per cent, failure rate 2·4 per cent). Conclusion Women in the UK had a disproportionate risk of admission without surgical intervention and had high rates of normal appendicectomy. Risk prediction models to support shared decision‐making by identifying adults in the UK at low risk of appendicitis were identified

Tensiones entre lo público, lo privado y lo estatal en tramas educativas situadas : IV Seminario taller Red de Antropología y Educación, 2018

La Red de Antropología y Educación (RIAE) surge por iniciativa de equipos de investigación radicados en el Área de Investigación Educativa del Centro de Estudios Avanzados de la Facultad de Ciencias Sociales y en el Centro de Investigaciones de la Facultad de Filosofía y Humanidades de la Facultad de Filosofía y Humanidades, Universidad Nacional de Córdoba, quienes en 2010 convocaron a sus pares de las Universidades Nacionales de Rosario y Buenos Aires al primer seminario – taller en Huerta Grande, Córdoba, con el objetivo de construir un espacio de intercambio de experiencias de investigación en el campo de la Antropología y Educación. Así, la RIAE permitió anudar múltiples trayectorias individuales y colectivas de investigadores formados y en formación interpelados por las profundas transformaciones sociales de las últimas décadas, como los procesos de desigualdad, los derroteros de las políticas públicas y siempre preocupados por reflexionar el lugar de “los sujetos” en dichos procesos y en el contexto de las propias investigaciones. En tal sentido, el Área de Investigación Educativa del CEA asumió un papel insustituible en la construcción de la RIAE y en la institucionalización del enfoque Socio-Antropológico Córdoba y en Argentina, a través de la Maestría en Investigación Educativa, el Postítulo en Investigación Educativa y el afianzamiento de línea Socio-Antropológica en el Doctorado en Estudios Sociales de América Latina del CEA. Es por ello que la publicación de los trabajos presentados en este IV Seminario-Taller de la RIAE en la Colección Actas de la Editorial del CEA significa una apuesta académica que busca impulsar una política de pesquisa sensible a las problemáticas sociales y educativas a la vez que portadora de una mirada crítica y fundada para atender nuestras responsabilidades como universitarios en los contextos actuales. Uno de los aspectos distintivos de la RIAE es que alienta la producción colectiva de conocimientos en el campo de Antropología y Educación. En tal sentido, promueve la presentación de trabajos de los equipos y no producciones individuales de sus miembros. Las producciones aquí reunidas respetan este criterio y presentan las reflexiones y debates de equipos de investigación radicados en Universidades Nacionales en distintos puntos del 8país que permiten explorar las formas que nos damos colectivamente para comprender lo social. Muchos esfuerzos personales e institucionales hicieron de la organización del IV Seminario-Taller un espacio de aprendizajes, debates y resistencias. Va nuestro agradecimiento al Área de Educación del Centro de Investigaciones, a la Escuela de Ciencias de la Educación, al Departamento de Antropología de la Facultad de Filosofía y Humanidades y al Área de Investigación Educativa del Centro de Estudios Avanzados, Facultad de Ciencias Sociales de la Universidad Nacional de Córdoba que procuraron las condiciones necesarias para el encuentro de tantas voluntades

Quinolone based chemosensor for the naked-eye and spectrophotometric detection of Cu2+in aqueous media

A new 4-hydroxy-2-quinolone based chemosensor (BPHTQ-1) was designed for the spectrophotometric and naked-eye detection of Cu2+. The chemosensor displayed a high selectivity and sensitivity towards Cu2+in the presence of other competitive metal cations in pure aqueous media. The Cu2+recognition furnished a distinguishable color change of BPHTQ-1 from colorless to yellow with a significant hyperchromic shift at 300 nm. The S and O atoms of BPHTQ-1 provided a 1:1 binding scaffold for the recognition of Cu2+ion with a high binding affinity of 19,338 M- 1and a detection limit of 1.39 μM, which is quite low compared with World Health Organization (WHO) reports.DST, New Delhi, India; Ministerio de Economia y Competitividad (Spain)/FEDER (EU) [BFU2011-30161-C02-02

Codon optimization of the adenoviral fiber negatively impacts structural protein expression and viral fitness

Codon usage adaptation of lytic viruses to their hosts is determinant for viral fitness. In this work, we analyzed the codon usage of adenoviral proteins by principal component analysis and assessed their codon adaptation to the host. We observed a general clustering of adenoviral proteins according to their function. However, there was a significant variation in the codon preference between the host-interacting fiber protein and the rest of structural late phase proteins, with a non-optimal codon usage of the fiber. To understand the impact of codon bias in the fiber, we optimized the Adenovirus-5 fiber to the codon usage of the hexon structural protein. The optimized fiber displayed increased expression in a non-viral context. However, infection with adenoviruses containing the optimized fiber resulted in decreased expression of the fiber and of wild-type structural proteins. Consequently, this led to a drastic reduction in viral release. The insertion of an exogenous optimized protein as a late gene in the adenovirus with the optimized fiber further interfered with viral fitness. These results highlight the importance of balancing codon usage in viral proteins to adequately exploit cellular resources for efficient infection and open new opportunities to regulate viral fitness for virotherapy and vaccine development.This work was supported by grants from the Spanish Ministry of Economia y Competitividad BIO2014-57716-C2-2-R and receives partial support from the Generalitat de Catalunya SGR14/248. CIBER de Enfermedades Raras is an initiative of the ISCIII. CF group is partially financed by the Instituto de Salud Carlos III (IIS10/00014) and co-financed by Fondo Europeo de Desarrollo Regional (FEDER). We also acknowledge the support of COST Action BM1204 EUPancreas. E.V. was supported by a fellowship from the Gobierno Vasco, Spain