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

The clerodane diterpene casearin J induces apoptosis of T-ALL cells through SERCA inhibition, oxidative stress, and interference with Notch1 signaling

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematologic malignancy that preferentially affects children and adolescents. Over 50% of human T-ALLs possess activating mutations of Notch1. The clerodane diterpene casearin J (CJ) is a natural product that inhibits the sarcoendoplasmatic reticulum calcium ATPase (SERCA) pump and induces cell death in leukemia cells, but the molecular mechanism of cytotoxicity remains poorly understood. Here we show that owing to SERCA pump inhibition, CJ induces depletion of the endoplasmic reticulum calcium pools, oxidative stress, and apoptosis via the intrinsic signaling pathway. Moreover, Notch1 signaling is reduced in T-ALL cells with auto-activating mutations in the HD-domain of Notch1, but not in cells that do not depend on Notch1 signaling. CJ also provoked a slight activation of NF-κB, and consistent with this notion a combined treatment of CJ and the NF-κB inhibitor parthenolide (Pt) led to a remarkable synergistic cell death in T-ALL cells. Altogether, our data support the concept that inhibition of the SERCA pump may be a novel strategy for the treatment of T-ALL with HD-domain-mutant Notch1 receptors and that additional treatment with the NF-κB inhibitor parthenolide may have further therapeutic benefits.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigaciones en Productos Naturales (CIPRONA)UCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Químic

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

Interventions and decision-making at the end of life: the effect of establishing the terminal illness situation

Background: Mobile learning (m-learning) has becoming very popular in education due to the rapidly advancing technology in our society. The potentials of the mobile applications should be used to enhance the education process. Few mobile applications have been designed to complement the study of physical therapy skills for physiotherapy students. The aim of this study was to investigate whether a mobile application, as a supplement to traditional learning, is useful for physiotherapy students in the acquisition of palpation and ultrasound skills in the shoulder area. Methods: Forty-nine students participated in this single-blinded, randomized controlled study. They were randomly distributed into two groups: experimental, with free access to the mobile application; and control, with access to traditional learning materials on the topic. Objective structured clinical evaluation (OSCE) and multiple-choice questionnaire (MCQ) were used to assess the educational intervention. Then, we also assessed the time taken to get a reliable ultrasound image and to localize a specific shoulder structure by palpation. Results: There was no significant intergroup difference in the acquisition of theoretical knowledge (p = .089). Scores were significantly higher in the experimental group than in the control group for the majority items in the ultrasound assessment; positioning of patient (p < .001), positioning of ultrasound probe (p = 0.007), handling of ultrasound probe (p = .013) and global OSCE (p < .001) and skills in palpation of the shoulder; position of patient (p = .009), direction of palpation contact (p = .021) and global OSCE (p = .034). There were no significant differences in the time required to perform the examination between groups in ultrasound (p = .944) and palpation (p = .393). The results from the post-program survey assessing the global satisfaction with the mobile application were high (8.200 ± .767), on an 11 numeric point rating scale. Conclusion: These results suggest the effectiveness of an m-learning program as a complement to traditional education for developing skills in ultrasound and palpation of the shoulder region in undergraduate physiotherapy students.The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This study was funded by a grant from the Educational Innovation Unit of the University of Granada, Spain (PID 13–86)

Vtil y breve institvtion para aprender los principios y fundamentos de la lengua Hespañola (1555)

PC 486; 8 Mb de memoria RAM; MS-DOS 6.0 y Microsoft Windows 3.1 o superior; Espacio libre superior a 10 Mb; Monitor VGA, resolución 800x600; Lector de CD-ROM local o en red.Tít. tomado de la etiquetaBello, Andrés: Gramática de la lengua castellana: destinada al uso de los americanos. Caracas: [s. n.] (V. Espinal), 1859. -- Benot, Eduardo: Arte de hablar: gramática filosófica de la lengua castellana. Madrid: [s. n.] (Librería de los Sucesores de Hernando), 1910. -- Calderón, Juan: Análisis lógica y gramatical de la lengua española. Madrid: [s. n.] (Imprenta de A. Vicente), 1852. -- Calleja, Juan Manuel: Elementos de gramática castellana. En Bilbao: por Don Pedro Antonio de Apraiz, 1818. -- Correas, Gonzalo: Arte grande de la lengua castellana.Madrid: [s. n.], 1903. -- Corro, Antonio De: The spanish grammer: with certeine rules teaching both the spanish and french tongues. Londres: por Iohn Wolse, 1590. -- Doergangk, Henrich: Institvtiones in lingvam hispanicam ad modvm faciles, qvales ante hac nunquam visae... Coloniae: Imprimebat Petrus à Brachel, 1614. -- Fernández Monje, Isidoro: Curso elemental de la lengua española. Madrid: [s. n.] (Librería de D. León Pablo Villaverde), 1854. -- Franciosini, Lorenzo: Grammatica spagnuola ed italiana. Genevra [Ginebra]: [s. n.], 1707. -- Gramática de la Lengua Vulgar de España. Lovaina: por Bartholomé Gravio, 1559. -- Gramática de la lengua castellana. Madrid: por la viuda de Don Joaquín Ibarra, 1796. -- Gramática de la lengua castellana. Madrid: [s. n.] (Imprenta Nacional), 1854. -- Gramática de la lengua castellana. Madrid: [s. n.] (Imprenta y Estereotipia de M. Rivadeneyra), 1870. -- Gramática de la lengua castellana.Madrid: Perlado, Páez y Compañía, 1920. -- Herrainz y De Heras, Gregorio: Tratado de gramática razonada: con aplicación decidida y constante al estudio del idioma español. Segovia: [s. n.] (Establecimiento Tipográfico de F. Santiuste), 1885. -- Jiménez Patón, Bartolomé: Institvciones de la gramatica española. Impresso en Baeza: por Pedro de la Cuesta, 1614. -- Jovellanos, Gaspar Melchor De: Curso de humanidades castellanas. [S.l.]: [s.n.], [s.d.]. -- Lancelot, Claude: Nouvelle methode pour apprendre facilement et en peu de temps la langue espagnole... París: Por Denys Thierry, 1681. -- Luna, Juan De: Arte breve, y conpendiossa para aprender a leer, escribir, pronunciar, y hablar la Lengua Española. Londres: por Iuan Guillermo, 1623. -- Martínez Gómez-Gayoso, Benito: Gramatica de la lengua castellana... En Madrid: en la imprenta de D. Gabriel Ramírez, 1769. -- Martínez López, Pedro: Principios de la lengua castellana ó prueba contra todos los que asienta D. Vicente Salvá en su gramática. Madrid: [s. n.] (librería de la señora viuda de Calleja é hijos), 1841. -- Meurier, Gabriel: Conjugaisons règles et instructions: mout propres et nécessairement requises pour ceux qui désirent apprendre françois, italien espagnol et flamen (1558). Anvers: Chez lan van Vvaesberghe, [1558]. -- Miranda, Juan De: Osservationi della lingva castigliana. In Vinegia: Gabriel Giolito de Ferrari, 1556. -- Nebrija, Elio Antonio De: Gramática de la lengua castellana. [S.l.]: [s.n.], [1492]. -- Noboa, A. M. De: Nueva gramática de la lengua castellana según los principios de la filosofía gramatical. Madrid: [s. n.] (Imprenta de Don Eusebio Aguado), 1839. -- Ovdin, Cesar: Grammaire espagnolle expliquée en françois. En Paris: Por Marc Orry, 1606. -- Perceval, Richard: Bibliotheca hispanica. En Londres: por Iohn Iackfon para Richard Watkins, 1591. -- Real Academia Española: Gramática de la lengua castellana. En Madrid: por Joachin de Ibarra, 1771. -- Real Academia Española: Gramática de la lengua castellana. Madrid: [s. n.] (Imprenta Nacional), 1858. -- Salleras, Matias: Gramática razonada de la lengua española. Segovia: [s. n.] (Imprenta de Pedro Ondero), 1876. -- Salvá, Vicente: Gramática de la lengua castellana según ahora se habla. París; Méjico: [s. n.], 1852. -- San Pedro, Benito De: Arte del romance castellano [2 tomos]. En Valencia: En la Imprenta de Benito Monfort, 1769. -- Sanford, John: An entrance to the spanish tongue. Londres: por Th. Haueland, 1611. -- Saqueniza, Jacobo: Gramática elemental de la lengua castellana: con un compendio de ortografía. Madrid: [s. n.] (Imprenta de Don Norberto Llorenci), 1832. -- Sobrino, Francisco: Gramatica nueva española y francesa. En Brusselas: Por Francisco Foppens, 1738. -- Stepney, William: The spanish schoole-master. Londres: por R. Field para Iohn Harison, 1591. -- Vtil y breve institvtion para aprender los principios y fundamentos de la lengua Hespañola. Lovanii: Ex officina Bartholomaei Graui, 1555. -- Villalón, Cristóbal De: Gramática Castellana: Arte breue y compendiosa para saber hablar y escreuir en la lengua Castellana congrua y decentemente. En Anvers: En casa de Guillermo Simon, 1558. -- Villar, Juan: Arte de la lengva española redvcida a reglas y preceptos de rigurosa gramatica. En Valencia: por Francisco Verengel, 1651

Novel genes and sex differences in COVID-19 severity.

Here we describe the results of a genome-wide study conducted in 11 939 COVID-19 positive cases with an extensive clinical information that were recruited from 34 hospitals across Spain (SCOURGE consortium). In sex-disaggregated genome-wide association studies for COVID-19 hospitalization, genome-wide significance (p &lt; 5x10-8) was crossed for variants in 3p21.31 and 21q22.11 loci only among males (p =&nbsp;1.3x10-22 and p =&nbsp;8.1x10-12, respectively), and for variants in 9q21.32 near TLE1 only among females (p =&nbsp;4.4x10-8). In a second phase, results were combined with an independent Spanish cohort (1598 COVID-19 cases and 1068 population controls), revealing in the overall analysis two novel risk loci in 9p13.3 and 19q13.12, with fine-mapping prioritized variants functionally associated with AQP3 (p =&nbsp;2.7x10-8) and ARHGAP33 (p =&nbsp;1.3x10-8), respectively. The meta-analysis of both phases with four European studies stratified by sex from the Host Genetics Initiative confirmed the association of the 3p21.31 and 21q22.11 loci predominantly in males and replicated a recently reported variant in 11p13 (ELF5, p = 4.1x10-8). Six of the COVID-19 HGI discovered loci were replicated and an HGI-based genetic risk score predicted the severity strata in SCOURGE. We also found more SNP-heritability and larger heritability differences by age (&lt;60 or ≥ 60&nbsp;years) among males than among females. Parallel genome-wide screening of inbreeding depression in SCOURGE also showed an effect of homozygosity in COVID-19 hospitalization and severity and this effect was stronger among older males. In summary, new candidate genes for COVID-19 severity and evidence supporting genetic disparities among sexes are provided