Met signaling in cardiomyocytes is required for normal cardiac function in adult mice

et al.Hepatocyte growth factor (HGF) and its receptor, Met, are key determinants of distinct developmental processes. Although HGF exerts cardio-protective effects in a number of cardiac pathologies, it remains unknown whether HGF/Met signaling is essential for myocardial development and/or physiological function in adulthood. We therefore investigated the requirement of HGF/Met signaling in cardiomyocyte for embryonic and postnatal heart development and function by conditional inactivation of the Met receptor in cardiomyocytes using the Cre-α-MHC mouse line (referred to as α-MHCMet-KO). Although α-MHCMet-KO mice showed normal heart development and were viable and fertile, by 6. months of age, males developed cardiomyocyte hypertrophy, associated with interstitial fibrosis. A significant upregulation in markers of myocardial damage, such as β-MHC and ANF, was also observed. By the age of 9. months, α-MHCMet-KO males displayed systolic cardiac dysfunction. Mechanistically, we provide evidence of a severe imbalance in the antioxidant defenses in α-MHCMet-KO hearts involving a reduced expression and activity of catalase and superoxide dismutase, with consequent reactive oxygen species accumulation. Similar anomalies were observed in females, although with a slower kinetics. We also found that Met signaling down-regulation leads to an increase in TGF-β production and a decrease in p38MAPK activation, which may contribute to phenotypic alterations displayed in α-MHCMet-KO mice. Consistently, we show that HGF acts through p38α to upregulate antioxidant enzymes in cardiomyocytes. Our results highlight that HGF/Met signaling in cardiomyocytes plays a physiological cardio-protective role in adult mice by acting as an endogenous regulator of heart function through oxidative stress control.This work was supported by grants: (AFM)-13683 from Association Française contre les myopathies, France, FIS-PI07/0071 and SAF-2010-20198-C02-01 from Ministry of Science and Innovation, Spain, and grants from Comunidad de Madrid/Universidad Complutense de Madrid: CAM/UCM 920384 and UCM-BSCH 920384, Spain to A.P.; BFU2011-25304 from Ministry of Science and Innovation, Spain, RD12/0019/0022 (TerCel network, ISCIII), P11-CTS-7564 (Junta de Andalucía) to R. M.-Ch.; FRM (Fondation pour la Recherche Médicale), Fondation Bettencourt-Schueller, and Association Française contre les myopathies (AFM) to F.M.; SAF2010-15881 from Ministry of Science and Innovation, Spain, and RD012/0021 (RedinRen network, ISCIII), and GR100 (Junta de Castilla y León) to J.M. L.-N. The cardiovascular phenotyping unit of the University of Salamanca, including the telemetry equipment, has been acquired with the support of the European Regional Development Funds (FEDER).Peer Reviewe

Chemical content and sensory changes of Oloroso Sherry wine when aged with four different wood types

Oloroso Sherry Wine is a fortified Sherry wine obtained by oxidation and ageing in American oak barrels of 500 L–600 L. In this work, the study of the suitability of other types of woods for the ageing of these wines was carried out. To compare the characteristics of the alternative woods, an oloroso wine was aged in four groups of 16 L barrels made of French oak, Spanish oak, chestnut, as well as American oak as control, with intense and medium toasting. Phenolic and furanic compounds, organic acids, volatile compounds, color characteristics, total polyphenol index and sensory analysis of wines aged for two months were analyzed. The results confirmed that the aged samples could be differentiated on the basis of their chemical composition, and that the use of alternative woods to age oloroso Sherry wines, and the level of wood toasting, had the potential to provide products with specific differences to the traditional aged in American oak. Furthermore, the organoleptic characteristics of these alternative wines were valued above a standard Sherry wine

Production yields at the distal fall-off of the β+ emitters 11C and 13N for in-vivo range verification in proton therapy

In proton therapy, Positron Emission Tomography (PET) range verification relies on the comparison of the measured and estimated activity distributions from β+ emitters produced by the proton beam in the patient. The accuracy of the estimated activity distributions is basically that of the underlying reaction cross section data. In this context, we have developed a new method for measuring β+ production yields combining the multi-foil technique with a clinical PET scanner, resulting in energy differential cross sections from a single irradiation. The method has been applied to the production of (t1/2 = 20.36 min) and (t1/2 = 9.97 min), the main candidates for off-line PET range verification, in carbon, nitrogen and oxygen, the main elements of the human body. The energy range studied with the 18 MeV CNA cyclotron corresponds to the distal fall-off of the activity curve, i.e. near the Bragg peak.Ministerio de Economía y Competitividad RYC-2014-15271, FPA2016- 77689-C2-1-R, RTI2018-098117-B-C2

Risk factors for thrombotic microangiopathy in allogeneic hematopoietic stem cell recipients receiving GVHD prophylaxis with tacrolimus plus MTX or sirolimus

Post-transplant complications.-- et al.Transplantation-associated thrombotic microangiopathy (TA-TMA) is a feared complication of allogeneic hematopoietic SCT (HSCT) owing to its high mortality rate. The use of calcineurin inhibitors or sirolimus (SIR) for GVHD prophylaxis has been suggested as a potential risk factor. However, the impact of tacrolimus (TAC) and SIR combinations on the increased risk of TA-TMA is currently not well defined. We retrospectively analyzed the incidence of TA-TMA in 102 allogeneic HSCT recipients who consecutively received TAC plus SIR (TAC/SIR) (n=68) or plus MTX (TAC/MTX)±ATG (n=34) for GVHD prophylaxis. No significant differences were observed in the incidence of TA-TMA between patients receiving TAC/SIR vs TAC/MTX±ATG (7.4% vs 8.8%, P=0.8). Only grade III-IV acute GVHD, previous HSCT and serum levels of TAC >25 ng/mL were associated with a greater risk of TA-TMA. Patients developing TA-TMA have significantly poorer survival (P<0.001); however, TA-TMA ceased to be an independent prognostic factor when it was included in a multivariate model. In conclusion, the combination of TAC/SIR does not appear to pose a higher risk of TA-TMA. By contrast, we identified three different risk groups for developing TA-TMA.Peer Reviewe

Organization and characterization of the promoter elements of the rRNA pperons in the slow-growing pathogen mycobacterium kumamotonense

The slow-growing, nontuberculous mycobacterium Mycobacterium kumamotonense possesses two rRNA operons, rrnA and rrnB, located downstream from the murA and tyrS genes, respectively. Here, we report the sequence and organization of the promoter regions of these two rrn operons. In the rrnA operon, transcription can be initiated from the two promoters, named P1 rrnA and PCL1, while in rrnB, transcription can only start from one, called P1 rrnB. Both rrn operons show a similar organization to the one described in Mycobacterium celatum and Mycobacterium smegmatis. Furthermore, by qRT-PCR analyses of the products generated from each promoter, we report that stress conditions such as starvation, hypoxia, and cellular infection affect the contribution of each operon to the synthesis of pre-rRNA. It was found that the products from the PCL1 promoter of rrnA play a pivotal role in rRNA synthesis during all stress conditions. Interestingly, the main participation of the products of transcription from the P1 promoter of rrnB was found during hypoxic conditions at the NRP1 phase. These results provide novel insights into pre-rRNA synthesis in mycobacteria, as well as the potential ability of M. kumamotonense to produce latent infection

Analysis of incidence, risk factors and clinical outcome of thromboembolic and bleeding events in 431 allogeneic hematopoietic stem cell transplantation recipients

This is an open-access paper.-- et al.Allogeneic hematopoietic stem cell transplantation recipients have an increasing risk of both hemorrhagic and thrombotic complications. However, the competing risks of two of these life-threatening complications in these complex patients have still not been well defined. We retrospectively analyzed data from 431 allogeneic transplantation recipients to identify the incidence, risk factors and mortality due to thrombosis and bleeding. Significant clinical bleeding was more frequent than symptomatic thrombosis. The cumulative incidence of a bleeding episode was 30.2% at 14 years. The cumulative incidence of a venous or arterial thrombosis at 14 years was 11.8% and 4.1%, respectively. The analysis of competing factors for venous thrombosis revealed extensive chronic graft-versus-host disease to be the only independent prognostic risk factor. By contrast, six factors were associated with an increased risk of bleeding; advanced disease, ablative conditioning regimen, umbilical cord blood transplantation, anticoagulation, acute III-IV graft-versus-host disease, and transplant-associated microangiopathy. The development of thrombosis did not significantly affect overall survival (P=0.856). However, significant clinical bleeding was associated with inferior survival (P<0.001). In allogeneic hematopoietic stem cell transplantation, significant clinical bleeding is more common than thrombotic complications and affects survival.Peer Reviewe

Characterization of the platelet phenotype caused by a germline RUNX1 Variant in a CRISPR/Cas9-generated murine model

RUNX1-related disorder (RUNX1-RD) is caused by germline variants affecting the RUNX1 gene. This rare, heterogeneous disorder has no specific clinical or laboratory phenotype, making genetic diagnosis necessary. Although international recommendations have been established to classify the pathogenicity of variants, identifying the causative alteration remains a challenge in RUNX1-RD. Murine models may be useful not only for definitively settling the controversy about the pathogenicity of certain RUNX1 variants, but also for elucidating the mechanisms of molecular pathogenesis. Therefore, we developed a knock-in murine model, using the CRISPR/Cas9 system, carrying the RUNX1 p.Leu43Ser variant (mimicking human p.Leu56Ser) to study its pathogenic potential and mechanisms of platelet dysfunction. A total number of 75 mice were generated; 25 per genotype (RUNX1WT/WT, RUNX1WT/L43S, and RUNX1L43S/L43S). Platelet phenotype was assessed by flow cytometry and confocal microscopy. On average, RUNX1L43S/L43S and RUNX1WT/L43S mice had a significantly longer tail-bleeding time than RUNX1WT/WT mice, indicating the variant's involvement in hemostasis. However, only homozygous mice displayed mild thrombocytopenia. RUNX1L43S/L43S and RUNX1WT/L43S displayed impaired agonist-induced spreading and α-granule release, with no differences in δ-granule secretion. Levels of integrin αIIbβ3 activation, fibrinogen binding, and aggregation were significantly lower in platelets from RUNX1L43S/L43S and RUNX1WT/L43S using phorbol 12-myristate 13-acetate (PMA), adenosine diphosphate (ADP), and high thrombin doses. Lower levels of PKC phosphorylation in RUNX1L43S/L43S and RUNX1WT/L43S suggested that the PKC-signaling pathway was impaired. Overall, we demonstrated the deleterious effect of the RUNX1 p.Leu56Ser variant in mice via the impairment of integrin αIIbβ3 activation, aggregation, α-granule secretion, and platelet spreading, mimicking the phenotype associated with RUNX1 variants in the clinical setting.This work was partially supported by grants from Instituto de Salud Carlos III (ISCIII) and Feder (PI17/01311, PI17/01966, and CB15/00055), Fundación Séneca (19873/GERM/15), Gerencia Regional de Salud (GRS 2061A/19 and 1647/A/17), Fundación Mutua Madrileña (FMM, AP172142019), and Sociedad Española de Trombosis y Hemostasia (SETH-FETH; Premio López Borrasca 2019 and Ayuda a Grupos de Trabajo en Patología Hemorrágica 2019). The authors' research on IPDs is conducted in accordance with the aims of the Functional and Molecular Characterization of Patients with Inherited Platelet Disorders Project, which is supported by the Hemorrhagic Diathesis Working Group of the Spanish Society of Thrombosis and Haemostasis. A.M.-Q., C.F.-I., and L.H.-C. were supported by predoctoral grants from the Junta de Castilla y León, Spain. E.V. was supported by the predoctoral grant from the University of Salamanca, Spain. IG-T and RB were supported by "Contratos postdoctorales Programa II) from the University of Salamanca, Spain

Searching for glycosylated natural products in actinomycetes and identification of novel macrolactams and angucyclines

Many bioactive natural products are glycosylated compounds in which the sugar components usually participate in interaction and molecular recognition of the cellular target. Therefore, the presence of sugar moieties is important, in some cases essential, for bioactivity. Searching for novel glycosylated bioactive compounds is an important aim in the field of the research for natural products from actinomycetes. A great majority of these sugar moieties belong to the 6-deoxyhexoses and share two common biosynthetic steps catalyzed by a NDP-D-glucose synthase (GS) and a NDP-D-glucose 4,6-dehydratase (DH). Based on this fact, seventy one Streptomyces strains isolated from the integument of ants of the Tribe Attini were screened for the presence of biosynthetic gene clusters (BGCs) for glycosylated compounds. Total DNAs were analyzed by PCR amplification using oligo primers for GSs and DHs and also for a NDP-D-glucose-2,3-dehydratases. Amplicons were used in gene disruption experiments to generate non-producing mutants in the corresponding clusters. Eleven mutants were obtained and comparative dereplication analyses between the wild type strains and the corresponding mutants allowed in some cases the identification of the compound coded by the corresponding cluster (lobophorins, vicenistatin, chromomycins and benzanthrins) and that of two novel macrolactams (sipanmycin A and B). Several strains did not show UPLC differential peaks between the wild type strain and mutant profiles. However, after genome sequencing of these strains, the activation of the expression of two clusters was achieved by using nutritional and genetic approaches leading to the identification of compounds of the cervimycins family and two novel members of the warkmycins family. Our work defines a useful strategy for the identification new glycosylated compounds by a combination of genome mining, gene inactivation experiments and the activation of silent biosynthetic clusters in Streptomyces strains

Didactic handbook in visual format fot the practice of specialized translation

La experiencia docente de los participantes en este proyecto ha puesto de relieve la importancia de orientar sus principios didácticos a fomentar la incentivación y el compromiso de los alumnos en el proceso de aprendizaje, así como la explicitación y ordenación de metodologías y materiales, adecuándolos a las necesidades y capacidades del alumno de traducción. De ahí, surge la iniciativa de proporcionar al alumno una metodología práctica bien definida, de carácter progresivo, y sustentada en modelos de análisis de discurso y de análisis traductológico. Según esto, este proyecto se orienta a la creación de material docente para la práctica de la traducción especializada. En concreto se ha diseñado un manual virtual cuyo contenido se presenta mediante vídeos explicativos. Este material audiovisual se ha alojado, junto a los textos, recursos bibliográficos, modelos de análisis, traducciones comentadas, etc., en la plataforma virtual EXA ENOA2, de acceso abierto a los alumnos de la Universidad de Córdoba y con formato html5, para poder acceder a ella desde cualquier dispositivo móvil. El manual, estructurado en cuatro itinerarios (jurídico-económico-administrativo, científico-técnico, humanístico-literario y audiovisual), ofrece un modelo de análisis discursivo específico que permite al alumno detectar y priorizar las especificidades de cada género textual y sus particularidades lingüísticas. Estos análisis se aplican a un corpus bilingüe de textos (FR>ES / EN>ES) seleccionados y explicados por los docentes, de los que se aporta además una traducción al español y su correspondiente análisis traductológico. Este manual proporciona además, a modo de anexo, una selección de recursos documentales, en formato electrónico, de carácter especializado (diccionarios, bases terminológicas, manuales de traducción especializada, etc.) que el alumno puede consultar de modo inmediato o descargarse en su ordenador para futuras consultas. Este material en soporte virtual de acceso abierto pretende configurarse como manual de autoaprendizaje de técnicas de traducción especializada y se constituye como complemento didáctico para el perfeccionamiento de las destrezas de traducción en áreas especializadas, así como la asimilación de una metodología práctica y funcional que permita al alumno optimizar las fases que configuran el proceso traslativo.The teaching experience of the members of this project emphasises the importance of directing teaching principles at encouraging students’ motivation and commitment within the learning process, as well as specifying and classifying methodologies and materials in order to adapt them to the necessities and capabilities of the translation students. This fact gives rise to the initiative to provide students a fully-designed, progressive and practical methodology, which is based on the models for discourse analysis and translation analysis. In this regard, the present project targets the elaboration of teaching materials for the practice of specialised translation. In particular, we have designed a virtual manual whose contents are presented through explanatory videos. These audiovisual contents, together with texts, bibliographical resources, analysis models and commented translations, have been hosted in the EXA ENOA2 virtual platform – with open-access to all students at the University of Córdoba – in html5 format, so it can be accessed from any mobile device. The manual includes four itineraries (legal-economical, scientific-technical, humanistic-literary, and audiovisual). It offers a specific discourse analysis model for each field, helping the student to detect and prioritize the characteristics of each textual genre and their linguistic features. These analysis models are applied to a bilingual corpora (FR/EN) of texts selected and explained by the teachers. Moreover, a Spanish translation and the corresponding translation analysis are provided. The manual also procures an annex including a selection of documentary resources in electronic format – such as dictionaries, termbases, specialized translation guides, etc. – that students can check immediately or download for future consults. This open-access virtual material aims to be a self-learning tool for specialised translation and to be constituted as a teaching complement for further acquisition of translation skills in specialised fields, as well as a practical and functional methodology that enables students to optimise every phase of the translation process

Análisis Del Efecto De La Adición De Fibra Cítrica Del Bagazo De La Naranja En Las Propiedades Nutrimentales Y Sensoriales De Un Embutido Y Determinación De La Calidad Microbiológica

The orange bagasse (Citrus sinensis) is a residue with a significant content of citrus fiber (FC); it is used as a functional ingredient in the development of new products. We consider it essential to know the effect of the nutritional and sensorial properties of FC. The objective of this research was to evaluate the effect of powdered the orange bagasse on the nutritional and sensorial properties of pork and beef sausage. The sausage was prepared by mixing the proportions of minced pork and beef with the corresponding FC, adding spices, orange juice and a solution of natural dye Bixa Orellana, after 24 hours of maturation was stuffed into cellulose casings. A randomized design with three repetitions was used. The treatments were formulations with 0, 5, 10 and 15% of FC respectively, named treatments E0, E5, E10 and E15. The chemical variables evaluated were: moisture content, ash, fat, protein (analyzed by the AOAC method, 1990), and total dietary fiber (by the methodology described by Asp et al., 1983 and Prosky et al, 1988); the microbiological variables: aerobic mesophiles, total and fecal coliforms, Salmonella and Staphylococcus aureus, detected and quantified with the methods of the corresponding Official Mexican Standards. The sensory variable was the level of liking which was evaluated in 128 consumers, and a commercial sausage called "EC" was used. As a comparison product. In E5, E10 and E15 the fat levels decreased (p &lt;0.05). The ash increased as a function of the addition of FC. Regarding protein, no differences were observed between E0 and E5. In the treatments, E10 and E 15 the protein decreased significantly. Total dietary fiber at E5, E10, and E15 ranged from 4.38 to 10.50%. The consumer test favored E0, E5, and EC. We conclude that the orange bagasse exerts a positive effect by decreasing fat and increasing total dietary fiber. Aleson-Carbonell et al. (2003) report similar behavior. It is concluded that it is feasible to add FC no higher than 5% as an alternative to enrich sausages