Randomized study of aprotinin and DDAVP to reduce postoperative bleeding after cardiopulmonary bypass surgery

BACKGROUND: Patients on cardiopulmonary bypass (CPB) have an increased susceptibility to postoperative bleeding. Previous reports using desmopressin acetate (DDAVP) for the prevention of postoperative bleeding have given contradictory results, whereas the protease inhibitor aprotinin has been shown to reduce blood loss after this type of surgery. This randomized study was performed to assess the efficacy of DDAVP versus aprotinin in the prevention of bleeding after CPB. METHODS AND RESULTS: One hundred nine of 122 eligible patients were randomized to four different groups: Group A (n = 28) received aprotinin starting with a bolus of 2 x 10(6) KIU followed by a continuous infusion of 0.5 x 10(6) KIU/h until the end of surgery; group B (n = 25) received of DDAVP 0.3 micrograms/kg i.v. on completion of CPB; group C (n = 28) received two doses of DDAVP, the first as in group B and an additional dose 6 hours after surgery; group D (n = 28) received no treatment. There was a marked reduction of postoperative blood loss either at 12 hours (P < .01) or 72 hours (P < .02) in the aprotinin group compared with all other groups, whereas no significant effect was observed in either of the two DDAVP regimens. A significant reduction in the amount of blood used was observed only in the aprotinin group (P < .01). Of the plasma fibrinolytic components assayed, there was a significant reduction of the fibrin degradation product generation in the aprotinin group (P < .001), whereas a significant systemic hyperfibrinolysis was observed in both DDAVP-treated groups and the control group. No side effects related to the study drugs were observed in any patient. CONCLUSIONS: Aprotinin inhibited fibrinolysis; this correlated with a significant reduction of postoperative blood loss and need for blood replacement after CPB. Neither one nor two doses of DDAVP had a beneficial effect. Aprotinin offers a better alternative than DDAVP in the prevention of bleeding after CPB

Shared apical sorting of anion exchanger isoforms AE2a, AE2b1, and AE2b2 in primary hepatocytes

AE2 (SLC4A2) is the member of the Na(+)-independent anion exchanger (AE) family putatively involved in the secretion of bicarbonate to bile. In humans, three variants of AE2 mRNA have been described: the full-length transcript AE2a (expressed from the upstream promoter in most tissues), and alternative transcripts AE2b(1) and AE2b(2) (driven from alternate promoter sequences in a tissue-restricted manner, mainly in liver and kidney). These transcripts would result in AE protein isoforms with short N-terminal differences. To ascertain their translation, functionality, and membrane sorting, we constructed expression vectors encoding each AE2 isoform fused to GFP at the C-terminus. Transfected HEK293 cells showed expression of functional GFP-tagged AE2 proteins, all three isoforms displaying comparable AE activities. Primary rat hepatocytes transfected with expression vectors and repolarized in a collagen-sandwich configuration showed a microtubule-dependent apical sorting of each AE2 isoform. This shared apical sorting is liver-cell specific, as sorting of AE2 isoforms was basolateral in control experiments on polarized kidney MDCK cells. Hepatocytic apical targeting of AE2 isoforms suggests that they all may participate in the canalicular secretion of bicarbonate to bile

Pancreatic cancer escape variants that evade immunogene therapy through loss of sensitivity to IFNgamma-induced apoptosis

Combined injections into experimental tumor nodules of adenovirus encoding IL-12 and certain chemokines are capable to induce immune-mediated complete regressions. In this study, we found that the combination of two adenoviruses, one encoding IL-12 and other MIP3alpha (AdCMVIL-12+AdCMVMIP3alpha) was very successful in treating CT-26-derived colon carcinomas. However, in experimental tumors generated from the pancreatic carcinoma cell line Panc02 such combined treatment induces 50% of macroscopic complete regressions, although local relapses within 1 week are almost constant. We derived cell lines from such relapsing tumors and found that experimental malignancies derived from their inoculum were not amenable to treatment in any case with AdCMVIL-12+AdCMVMIP-3alpha. Importantly, relapsing cell lines were insensitive to in vitro induction of apoptosis by IFNgamma, in clear contrast with the original Panc02 cells. Comparative analyses by cDNA arrays of relapsing cell lines versus wild-type Panc02 were performed revealing an important number of genes (383) whose expression levels were modified more than two-fold. These changes grouped in certain gene ontology categories should harbor the mechanistic explanations of the acquired selective resistance to IFNgamma

Eliminating a Region of Respiratory Syncytial Virus Attachment Protein Allows Induction of Protective Immunity without Vaccine-enhanced Lung Eosinophilia

In a murine model of respiratory syncytial virus disease, prior sensitization to the attachment glycoprotein (G) leads to pulmonary eosinophilia and enhanced illness. Three different approaches were taken to dissect the region of G responsible for enhanced disease and protection against challenge. First, mutant viruses, containing frameshifts that altered the COOH terminus of the G protein, were used to challenge mice sensitized by scarification with recombinant vaccinia virus (rVV) expressing wild-type G. Second, cDNA expressing these mutated G proteins were expressed by rVV and used to vaccinate mice before challenge with wild-type respiratory syncytial virus (RSV). These studies identified residues 193–205 to be responsible for G-induced weight loss and lung eosinophilia and showed that this region was not was not necessary for induction of protective immunity. Third, mice were sensitized using an rVV that expressed only amino acids 124–203 of the G protein. Upon RSV challenge, mice sensitized with this rVV developed enhanced weight loss and eosinophilia. This is the first time that a region within RSV (amino acids 193–203) has been shown to be responsible for induction of lung eosinophilia and disease enhancement. Moreover, we now show that it is possible to induce protective immunity with an altered G protein without inducing a pathological response

Flexible workflows for on-the-fly electronmicroscopy single-particle image processing using Scipion

Electron microscopy of macromolecular structures is an approach that is in increasing demand in the field of structural biology. The automation of image acquisition has greatly increased the potential throughput of electron microscopy. Here, the focus is on the possibilities in Scipion to implement flexible and robust image-processing workflows that allow the electron-microscope operator and the user to monitor the quality of image acquisition, assessing very simple acquisition measures or obtaining a first estimate of the initial volume, or the data resolution and heterogeneity, without any need for programming skills. These workflows can implement intelligent automatic decisions and they can warn the user of possible acquisition failures. These concepts are illustrated by analysis of the well known 2.2 Å resolution β-galactosidase data setThe authors would like to acknowledge financial support from The Spanish Ministry of Economy and Competitiveness through the BIO2016-76400-R (AEI/FEDER, UE) grant, the Comunidad Auto´noma de Madrid through grant S2017/BMD3817, the Instituto de Salud Carlos III (PT17/0009/0010), the European Union (EU) and Horizon 2020 through the CORBEL grant (INFRADEV-1-2014-1, Proposal 654248), the ‘la Caixa’ Foundation (ID 100010434, Fellow LCF/BQ/ IN18/11660021), Elixir–EXCELERATE (INFRADEV-3- 2015, Proposal 676559), iNEXT (INFRAIA-1-2014-2015, Proposal 653706), EOSCpilot (INFRADEV-04-2016, Proposal 739563) and INSTRUCT–ULTRA (INFRADEV03-2016-2017, Proposal 731005

Un proyecto europeo para la gestión integral de las especies vegetales invasoras en la Costa Brava

Póster presentado en el III Simposio Anual de Botánica Española celebrado en el Institut Botànic de Barcelona, Barcelona, 25-26 de noviembre de 2022LIFE medCLIFFS es un proyecto financiado por el programa LIFE de la Unión Europea, con una duración de cinco años y que se focaliza en la Costa Brava y en el Parque Natural de Cap de Creus, una de las zonas con acantilados litorales más afectadas en Cataluña por la problemática de las especies vegetales invasoras. Las actividades del proyecto incluyen desde acciones preventivas a acciones de erradicación y, con el mismo nivel de importancia, acciones de concienciación social y de difusión. En este sentido, involucra administraciones públicas, científicos, voluntarios y sector productivo entre sus socios, y busca la colaboración de personas individuales y entidades locales. Las acciones se centran básicamente en la detección temprana y erradicación de cinco táxones —Opuntia ficus-indica y O. stricta, Carpobrotus acinaciformis y C. edulis (y sus posibles híbridos) y Gazania rigens— lo que debería también servir para reducir su impacto en áreas sensibles que alberguen especies endémicas amenazadas como Limonium geronense, L. tremolsii y Seseli farrenyi, únicas en el mundo. En paralelo, se han creado redes participativas para la detección temprana y control de la flora alóctona invasora o potencialmente invasora, integradas por observadores y voluntarios que aportan una información clave para actualizar y calibrar un sistema automático de evaluación del riesgo de invasión (RISKMAPR). Además, puesto que la jardinería y el uso de plantas ornamentales son la principal causa del establecimiento de plantas invasoras, están previstas también las siguientes acciones específicas: redacción de una guía de buenas prácticas, incluyendo una lista con las especies inocuas y la flora ornamental a evitar y creación de una etiqueta de calidad para los proveedores y grandes usuarios, como los ayuntamientos, entre otros

LIFE medCLIFFS: un proyecto europeo para la gestión integral de las especies vegetales invasoras en el litoral del nororiente de Cataluña (España)

Póster presentado en el V Congreso Ecuatoriano de Mastozoología, III Congreso Nacional de Manejo de Vida Silvestre y I Congreso Ecuatoriano de Centros de Rescate, Zoológicos, Acuarios y Afines, efectuados entre el 19 y 21 de octubre de 2022, en la ciudad Samborondón, GuayasLIFE medCLIFFS es un proyecto financiado por el programa LIFE de la Unión Europea, con una duración de cinco años y que focalizará sus actuaciones en la Costa Brava y el Parque Natural del Cap de Creus, una de las zonas con acantilados litorales más afectadas en Cataluña por la problemática de las especies vegetales invasoras. Este proyecto es pionero en la región de actuación (SO de Europa) por su visión integradora de la gestión de las especies invasoras: las actividades del proyecto incluyen desde acciones preventivas a acciones de erradicación, pasando por la detección temprana de especies y, con el mismo nivel de importancia, acciones de concienciación social/difusión e incluso acciones legales. En este sentido, involucra administraciones públicas, científicos, voluntarios y sector productivo entre sus socios, y busca la colaboración de personas individuales y entidades locales. Las acciones para controlar y erradicar especies alóctonas ya establecidas se centrarán básicamente en cinco táxones —Opuntia ficus-indica y O. stricta, Carpobrotus acinaciformis y C. edulis (y sus posibles híbridos), y Gazania rigens— lo que debería también servir para reducir su impacto en áreas sensibles que alberguen especies endémicas amenazadas como Limonium geronense, L. tremolsii y Seseli farrenyi, únicas en el mundo. En paralelo, se crearán redes participativas para la detección temprana y control de la flora alóctona invasora o potencialmente invasora, integradas por observadores y voluntarios que aportarán una información clave para actualizar y calibrar un sistema automático de evaluación del riesgo de invasión (RISKMAPR o similar). El proyecto incidirá, además, a través de campañas de educación y divulgación en el propio Parque Natural y otros sitios turísticos costeros del NE de Cataluña (Costa Brava), en la concienciación social sobre los efectos nocivos de estas especies invasoras y la necesidad de reducir su propagación. Además, en esta línea, y puesto que la jardinería y el uso de plantas ornamentales son la principal causa de su establecimiento, están previstas también las siguientes acciones específicas (1) redacción de una guía de buenas prácticas, incluyendo una lista con las especies inocuas y la flora ornamental a evitar; (2) creación de una etiqueta de calidad para los proveedores (viveros, empresas de jardinería y cadenas de supermercados, entre otros) y (3) redacción de un reglamento para el uso de flora ornamental a nivel municipal y aprobación en los municipios costeros de la costa Brava

LIFE medCLIFFS: un proyecto europeo para la gestión integral de las especies vegetales invasoras en el litoral del NE de Cataluña

Comunicación oral presentada en el XI Congreso de Biología de la Conservación de Plantas celebrado en Gran Canaria del 17 al 21 de julio de 202

Strategies to design clinical studies to identify predictive biomarkers in cancer research

The discovery of reliable biomarkers to predict efficacy and toxicity of anticancer drugs remains one of the key challenges in cancer research. Despite its relevance, no efficient study designs to identify promising candidate biomarkers have been established. This has led to the proliferation of a myriad of exploratory studies using dissimilar strategies, most of which fail to identify any promising targets and are seldom validated. The lack of a proper methodology also determines that many anti-cancer drugs are developed below their potential, due to failure to identify predictive biomarkers. While some drugs will be systematically administered to many patients who will not benefit from them, leading to unnecessary toxicities and costs, others will never reach registration due to our inability to identify the specific patient population in which they are active. Despite these drawbacks, a limited number of outstanding predictive biomarkers have been successfully identified and validated, and have changed the standard practice of oncology. In this manuscript, a multidisciplinary panel reviews how those key biomarkers were identified and, based on those experiences, proposes a methodological framework—the DESIGN guidelines—to standardize the clinical design of biomarker identification studies and to develop future research in this pivotal field