Relevance of p53 in the regulation of pro and antiapoptotic factors from the Bcl-2 family during the treatment with tirosine kinase inhibitors

Motivation: Liver cancer is the sixth most common cancer and the fourth most frequent cause of cancer-related death worldwide [1]. Nowadays, only one third of patients diagnosed with HCC are in the earliest BCLC 0-A stages, with a high proportion being in more advanced stages of the disease (BCLC B-C). Patients with the presence of poor prognostic factors such as vascular invasion, extrahepatic metastases and/or impaired hepatic function are considered in the advanced stage of the disease (BCLC C). Sorafenib is the standard of care for advanced HCC stage demonstrated in two large-scale trials [2]. Other drugs have been developed to increase the therapeutic arsenal of treatments. A phase III clinical trial comparing Lenvatinib and Sorafenib demonstrated that Lenvatinib was statistically non-inferior to Sorafenib in overall survival as a first-line treatment in patients with advanced HCC. Regorafenib and Cabozantinib have been shown to be effective as second-line therapy. [2]   Methods: Sorafenib, Regorafenib, Cabozantinib and Lenvatinib were obtained commercially from Carbosynth Limited (Berkshire, UK). HepG2, Hep3B and Huh7 cell lines were obtained from American Type Culture Collection (ATCC; LGC Standards, S.L.U., Barcelona, Spain). HCC cell lines were maintained in supplemented Minimum Essential Medium with Earle's Balanced Salts (MEM/EBSS) at 37°C in a humidified incubator with 5 % CO2. Cells were seeded at a density of 100,000 cells/cm2 in 2D culture. Different parameters related to cell death and proliferation were associated with the expression of proapoptotic (Bak, Bax, tBid and Bim) antiapoptotic (Bcl-2, Mcl-1 and Bcl-xL) and regulatory (Beclin-1) Bcl-2 family members assessed by Western-blot analysis.   Results: The administration of tyrosine kinase inhibitors induced cell death and reduced cell proliferation. This effect was associated with an upregulation of tBid and Bim expression in liver cancer cells. This effect was not observed in Hep3B and Huh7 which were less responsiveness to the proapoptotic and antiproliferative properties of tyrosine kinase inhibitors.   Conclusions: The induction of cell death and antiproliferative properties of tyrosine kinase inhibitors were associated with the increase of the expression of different proapoptotic Bcl-2 family members. This expression appeared to be regulated by p53

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

EFECTO DEL SULFATO DE AMONIO EN LA EFICIENCIA DEL HERBICIDA GLYPHOSATE PARA EL CONTROL DEL YSYPO´I

Con el objetivo de evaluar el efecto de sulfato de amonio en la eficiencia del herbicida Glyphosate para el control de Ysypo'i (/pomoea grandifo/ia L.), se realizó el presente trabajo en el Distrito La Paz, departamento de Itapúa.Se inició en Diciembre de 2007 y terminó en abril de 2008. Se utilizó el diseño completamente al azar con cuatro repeticiones. Las unidades experimentales estuvieron constituidas por cinco plantas de Ysypo'i (/pomoea grandifo/ia L.). Los tratamientos fueron conformados por el factorial de 2x3x3 dos épocas de aplicación en fase vegetativa, plantas con 4 a 6 hojas y fase reproductiva, tallo voluble desarrollado, tres dosis de glyphosate ( 2,0, 4,0 Y 6,0 I p.c./ha) y tres dosis de sulfato de amonio ( 0,2,0% Y 4,0%), Lo cual totaliza un número de 18 tratamientos.Este ensayo fue realizado en condiciones de campo. La pulverización fue realizada con mochila, en forma manual, con boquilla 90,03, temperatura 25aC, HR 70%. Las evaluaciones fueron realizadas a los 15, 30 Y 45 días después de la aplicación. Los resultados fueron expresados en porcentaje de control. La adición del sulfato de amonio al 2,0% (p/v) aumentó la eficiencia de la dosis más baja de g/yphosate 2,0 I p.c./ha. Que alcanzo un nivel excelente de control, 95%. Las dosis de 4,0 I.p.c. / ha de glyphosate sin adyuvante tuvo un excelente control ( 100%) de Ysypo'i en la fase vegetativa mas no en la fase reproductiva que sólo alcanzó 75%. La dosis de 6, 0 I p.c./há de g/yphosate sí tuvo un control excelente (100%), tanto en la etapa vegetativa como en la reproductiva. La adición de sulfato de amonio no aumentó la eficiencia de las dosis altas de g/yphosate, sobre todo en la fase reproductiva, que por el contrario, tuvo un efecto antagónico

Universal primers for the amplification and sequence analysis pf actin-1 from diverse mosquito species

We report the development of universal primers for the reverse-transcription polymerase chain reaction (RT-PCR) amplification and nucleotide sequence analysis of actin cDNAs from taxonomically diverse mosquito species. Primers specific to conserved regions of the invertebrate actin-1 gene were designed after actin cDNA sequences of Anopheles gambiae, Bombyx mori, Drosophila melanogaster, and Caenorhabditis elegans. The efficacy of these primers was determined by RT-PCR with the use of total RNA from mosquitoes belonging to 30 species and 8 genera (Aedes, Anopheles, Culex, Deinocerites, Mansonia, Psorophora, Toxorhynchites, and Wyeomyia). The RT-PCR products were sequenced, and sequence data were used to design additional primers. One primer pair, denoted as Act-2F (5′-ATGGTCGGYATGGGNCAGAAGGACTC-3′) and Act-8R (5′-GATTCCATACCCAGGAAG-GADGG-3′), successfully amplified an RT-PCR product of the expected size (683-nt) in all mosquito spp. tested. We propose that this primer pair can be used as an internal control to test the quality of RNA from mosquitoes collected in vector surveillance studies. These primers can also be used in molecular experiments in which the detection, amplification or silencing of a ubiquitously expressed mosquito housekeeping gene is necessary. Sequence and phylogenetic data are also presented in this report

Regulation of Fission Yeast Morphogenesis by PP2A Activator pta2

Cell polarization is key for the function of most eukaryotic cells, and regulates cell shape, migration and tissue architecture. Fission yeast, Schizosaccharomyces pombe cells are cylindrical and polarize cell growth to one or both cell tips dependent on the cell cycle stage. Whereas microtubule cytoskeleton contributes to the positioning of the growth sites by delivering polarity factors to the cell ends, the Cdc42 GTPase polarizes secretion via actin-dependent delivery and tethering of secretory vesicles to plasma membrane. How growth is restricted to cell tips and how re-initiation of tip growth is regulated in the cell cycle remains poorly understood. In this work we investigated the function of protein phosphatase type 2A (PP2A) in S. pombe morphogenesis by deleting the evolutionary conserved PTPA-type regulatory subunit that we named pta2. pta2-deleted cells showed morphological defects and altered growth pattern. Consistent with this, actin patches and active Cdc42 were mislocalized in the pta2 deletion. These defects were additive to the lack of Cdc42-GAP Rga4. pta2Δ cells show upregulated Cdc42 activity and pta2 interacts genetically with polarisome components Tea1, Tea4 and For3 leading to complete loss of cell polarity and rounded morphology. Thus, regulation of polarity by PP2A requires the polarisome and involves Pta2-dependent control of Cdc42 activity

A far-ultraviolet-driven photoevaporation flow observed in a protoplanetary disk.

Most low-mass stars form in stellar clusters that also contain massive stars, which are sources of far-ultraviolet (FUV) radiation. Theoretical models predict that this FUV radiation produces photodissociation regions (PDRs) on the surfaces of protoplanetary disks around low-mass stars, which affects planet formation within the disks. We report James Webb Space Telescope and Atacama Large Millimeter Array observations of a FUV-irradiated protoplanetary disk in the Orion Nebula. Emission lines are detected from the PDR; modeling their kinematics and excitation allowed us to constrain the physical conditions within the gas. We quantified the mass-loss rate induced by the FUV irradiation and found that it is sufficient to remove gas from the disk in less than a million years. This is rapid enough to affect giant planet formation in the disk

Promoción de la salud y entornos saludables

A forestar forestalAplicación de un programa educativo participativo en salud  bucal a una comunidad de adultos mayoresBiblioteca móvil y su implementación en el hospital Padre HurtadoConsumo de riesgo de alcohol en Chile: una propuesta innovadora de intervenciónDiseño de un programa interactivo de promoción de la salud vocal para NB1Encuentro formativo en promoción de salud y gestión de entornos saludables para TenoExperiencia docente: programa intersectorial de promoción/prevención en preescolares de comunas vulnerables, Región MetropolitanaFiltrado glomerular, método preventivo aparición de fibrosis sistémica nefrogénica por gadolinio en examen de RMImplementación de consejerías en vida sana en APS, Región de los RíosMedicina preventiva en feria libre de la población San Gregorio: Cecof San Gregorio, Contagiando SaludMetodología innovadora en la enseñanza de una ectoparasitosisPrevención de accidentes por monóxido de carbono en edificios, Providencia 2002-2009Programa de promoción y prevención en salud bucal para preescolaresPromoviendo hábitos saludables en los vecinos de Reñaca Alto, Viña del Mar, 2009Rol de la capacitación en la implementación de acciones para la prevención de la obesidadSatisfacción usuaria en el Cesfam Natales a un año de su funcionamientoTres estrategias publicitarias y de comunicación aplicadas al consumo de alcohol de bajo riesgoTropa de la salud: uso de los medios como forma de promover la salu

Global economic burden of unmet surgical need for appendicitis

Background: There is a substantial gap in provision of adequate surgical care in many low-and middle-income countries. This study aimed to identify the economic burden of unmet surgical need for the common condition of appendicitis. Methods: Data on the incidence of appendicitis from 170 countries and two different approaches were used to estimate numbers of patients who do not receive surgery: as a fixed proportion of the total unmet surgical need per country (approach 1); and based on country income status (approach 2). Indirect costs with current levels of access and local quality, and those if quality were at the standards of high-income countries, were estimated. A human capital approach was applied, focusing on the economic burden resulting from premature death and absenteeism. Results: Excess mortality was 4185 per 100 000 cases of appendicitis using approach 1 and 3448 per 100 000 using approach 2. The economic burden of continuing current levels of access and local quality was US $92 492 million using approach 1 and$73 141 million using approach 2. The economic burden of not providing surgical care to the standards of high-income countries was $95 004 million using approach 1 and$75 666 million using approach 2. The largest share of these costs resulted from premature death (97.7 per cent) and lack of access (97.0 per cent) in contrast to lack of quality. Conclusion: For a comparatively non-complex emergency condition such as appendicitis, increasing access to care should be prioritized. Although improving quality of care should not be neglected, increasing provision of care at current standards could reduce societal costs substantially

Measurement of hadronic event shapes in high-p T multijet final states at √s = 13 TeV with the ATLAS detector

A measurement of event-shape variables in proton-proton collisions at large momentum transfer is presented using data collected at s = 13 TeV with the ATLAS detector at the Large Hadron Collider. Six event-shape variables calculated using hadronic jets are studied in inclusive multijet events using data corresponding to an integrated luminosity of 139 fb−1. Measurements are performed in bins of jet multiplicity and in different ranges of the scalar sum of the transverse momenta of the two leading jets, reaching scales beyond 2 TeV. These measurements are compared with predictions from Monte Carlo event generators containing leading-order or next-to-leading order matrix elements matched to parton showers simulated to leading-logarithm accuracy. At low jet multiplicities, shape discrepancies between the measurements and the Monte Carlo predictions are observed. At high jet multiplicities, the shapes are better described but discrepancies in the normalisation are observed. [Figure not available: see fulltext.