Characterisation of PAMP/PRR interactions in European eel (Anguilla anguilla) macrophage-like primary cell cultures

The eel (Anguilla anguilla) has been identified as a vulnerable species with stocks dramatically declining over the past decade. In an effort to support the species from overfishing of wild stocks increased interest in eel aquaculture has been notable. In order to expand the scarce knowledge concerning the biology of this species significant research efforts are required in several fields of biology. The development of cell culture systems to study the immune response is a key step towards an increased understanding of the immune response and to develop resources to support further study in this threatened species. Macrophages are one of the most important effector cells of the innate immune system. The capacity to engulf pathogens and orchestrate the immune response relies on the existence of different surface receptors, such as scavenger receptors and toll-like receptors. We have developed and described an eel macrophage-like in vitro model and studied its functional and transcriptomic responses. Macrophagelike cells from both head kidney and purified peripheral blood leukocytes were obtained and phagocytic activity measured for different whole bacteria and yeast. Moreover, based on PAMP-PRR association the innate immune response of both head kidney and PBL derived macrophage-like cells was evaluated against different pathogen-associated molecular patterns (PAMPs). Results highlight that peptidoglycan stimulation strongly induces inflammatory mRNA expression reflected in the up-regulation of proinflammatory genes IL1b and IL18 in PBL derived cells whereas IL8 is upregulated in head kidney derived cells. Furthermore TLR2 mRNA abundance is regulated by all stimuli supporting a multifunctional role for this pathogen recognition receptor (PRR) in eel macrophage-like cells

Eel immune response to Vibrio vulnificus infection. Host-pathogen relationship

The European eel (Anguilla anguilla), has experienced a dangerous decline in recruitment, yield and stock over the last 30 years and this decline is likely to continue into the future. Several major threats are responsible for this situation, including overfishing of glass eels for consumption, new infections by introduced pathogens, dams and blocking of migration routes. The lack of knowledge concerning the biology of this species represents a handicap to the conservation and recovery of the population. Nevertheless,the eel farming industry is increasing, therefore it could be considered one of the major risks for wild eel stock perpetuation, which is threatened principally by the lack of knowledge of the complete reproductive cycle of the European eel and by emerging bacterial, viral and parasite diseases. Vibrio vulnificus is the aetiological agent of warm-water vibriosis, a disease that constitutes the main threat to eels under culture conditions, provoking important outbreaks and can be an opportunistic pathogen for humans. V. vulnificus uses a novel characterized virulence and survival system namedMARTX (multifunctional repeat in toxin) that it is supposed to trigger a cytokine storm when it is produced in blood during the infection. It is also involved in the colonization of the gills, to invade internal organs and causes death by septicemia. We sequenced an eel immune-enriched transcriptome with Roche 454, and the data has been used to create new molecular tools for further research. We have also designed a custom eel-specific microarray (4x44K, Agilent). Our research has been focused on eel-vibrio interaction, how the host responds to the challenge and the importance of MARTX system of the bacteria for the colonization. Different challenges were performed using wild type strain (CECT4999) and Rtx double mutant strain (CT285) to evaluate mucosal immunity in the gills, the principal portal of entry.We also studied the early immune response of circulating leukocytes and the role of the erythrocytes, as one of the target cells of the lytic activity of this bacterium. Results obtained by RT-qPCR and ISH on gills demonstrate the immunocompetent function of this tissue, based on the expression and localization of immune-relates genes such as, TLRs, cytokines, chemokines and signalling molecules. Microarray studies describe key genes expressed after vibrio infection analyzed across 12h post-challenge on gills and blood (leukocytes and erythrocytes separately), specific host response against vibrio Rtx toxin, as well as, a possible function of erythrocytes in the immune response

Electric vehicle penetration, comparison between Norway, Japan and Spain.

The aim of the study is to analyze how the country with the highest percentage of electric vehicles as is Norway has managed to become it and analyze the case of Japan and Spain with the charact of each country. The batteries are evolving and increasing their performance, but it is necessary a safe network of charging stations for electric vehicles to reduce the "range anxiety". A discuss of the electric car penetration with different Policies and Legislacy to incentivize the electric vehicle will be made to see the Spanish situation respect to Norway and Japan

An Enriched European Eel Transcriptome Sheds Light upon Host-Pathogen Interactions with Vibrio vulnificus

Infectious diseases are one of the principal bottlenecks for the European eel recovery. The aim of this study was to develop a new molecular tool to be used in host-pathogen interaction experiments in the eel. To this end, we first stimulated adult eels with different pathogen-associated molecular patterns (PAMPs), extracted RNA from the immune-related tissues and sequenced the transcriptome. We obtained more than 2 x 10(6) reads that were assembled and annotated into 45,067 new descriptions with a notable representation of novel transcripts related with pathogen recognition, signal transduction and the immune response. Then, we designed a DNA-microarray that was used to analyze the early immune response against Vibrio vulnificus, a septicemic pathogen that uses the gills as the portal of entry into the blood, as well as the role of the main toxin of this species (RtxA13) on this early interaction. The gill transcriptomic profiles obtained after bath infecting eels with the wild type strain or with a mutant deficient in rtxA13 were analyzed and compared. Results demonstrate that eels react rapidly and locally against the pathogen and that this immune-response is rtxA13-dependent as transcripts related with cell destruction were highly up-regulated only in the gills from eels infected with the wild-type strain. Furthermore, significant differences in the immune response against the wild type and the mutant strain also suggest that host survival after V. vulnificus infection could depend on an efficient local phagocytic activity. Finally, we also found evidence of the presence of an interbranchial lymphoid tissue in European eel gills although further experiments will be necessary to identify such tissue

Metodologia para a formação da competência obstétrica em estudantes de Enfermaria

The purpose of this article was to develop a methodology for the formation of obstetric competence aimed at Nursing students, which allows them to perform efficiently at different levels of health. Analytical-synthetic, inductive-deductive methods were used to process the information, and systemic to develop the proposed methodology. This was submitted to the consideration of specialists in work sessions organized in the form of critical opinion and collective construction workshops. As a result, a methodology was developed for the formation of obstetric competence based on the relationship between knowledge and procedures for the use of the academic network as a form of organization of teaching, it contains four stages: diagnosis of the state of the formation of obstetric competence, preparation of the conditions for the operation of the Maternal Health Nursing Network, development of training activities in the network and evaluation of student performance. The objectives, procedures and actions to be developed are described. The methodology designed for the training of obstetric competence through the use of the academic network as a form of organization is characterized by its flexibility and is tempered to the different training contexts. Its functionality will allow students to appropriate knowledge for efficient performance in solving professional problems through proper management in the teaching-learning process of training activities in virtual modality for the formation of obstetric competence.O objetivo deste artigo foi desenvolver uma metodologia para a formação de competências obstétricas destinada a estudantes de Enfermagem, que lhes permita atuar com eficiência em diferentes níveis de saúde. Métodos analítico-sintético, indutivo-dedutivo foram utilizados para processar as informações e sistêmicos para desenvolver a metodologia proposta. Este foi submetido à apreciação de especialistas em sessões de trabalho organizadas sob a forma de oficinas de opinião crítica e de construção coletiva. Como resultado, desenvolveu-se uma metodologia para a formação da competência obstétrica baseada na relação entre saberes e procedimentos para utilização da rede acadêmica como forma de organização do ensino, contém quatro etapas: diagnóstico do estado da formação da competência obstétrica, preparação das condições para o funcionamento da Rede de Enfermagem de Saúde Materna, desenvolvimento de atividades formativas na rede e avaliação do desempenho dos estudantes. Descrevem-se os objetivos, procedimentos e ações a serem desenvolvidas. A metodologia desenhada para o treino da competência obstétrica através da utilização da rede académica como forma de organização caracteriza-se pela sua flexibilidade e ajusta-se aos diferentes contextos de formação. Sua funcionalidade permitirá aos estudantes apropriar-se do conhecimento para um desempenho eficiente na resolução de problemas profissionais por meio de um gerenciamento adequado no processo ensino-aprendizagem das atividades de treinamento na modalidade virtual para a formação da competência obstétrica.O objetivo deste artigo foi desenvolver uma metodologia para a formação de competências obstétricas destinada a estudantes de Enfermagem, que lhes permita atuar com eficiência em diferentes níveis de saúde. Métodos analítico-sintético, indutivo-dedutivo foram utilizados para processar as informações e sistêmicos para desenvolver a metodologia proposta. Este foi submetido à apreciação de especialistas em sessões de trabalho organizadas sob a forma de oficinas de opinião crítica e de construção coletiva. Como resultado, desenvolveu-se uma metodologia para a formação da competência obstétrica baseada na relação entre saberes e procedimentos para utilização da rede acadêmica como forma de organização do ensino, contém quatro etapas: diagnóstico do estado da formação da competência obstétrica, preparação das condições para o funcionamento da Rede de Enfermagem de Saúde Materna, desenvolvimento de atividades formativas na rede e avaliação do desempenho dos estudantes. Descrevem-se os objetivos, procedimentos e ações a serem desenvolvidas. A metodologia desenhada para o treino da competência obstétrica através da utilização da rede académica como forma de organização caracteriza-se pela sua flexibilidade e ajusta-se aos diferentes contextos de formação. Sua funcionalidade permitirá aos estudantes apropriar-se do conhecimento para um desempenho eficiente na resolução de problemas profissionais por meio de um gerenciamento adequado no processo ensino-aprendizagem das atividades de treinamento na modalidade virtual para a formação da competência obstétrica

Analysis of the Adherence and Safety of Second Oral Glucose-Lowering Therapy in Routine Practice From the Mediterranean Area : A Retrospective Cohort Study

Altres ajuts: AstraZeneca/ESR-16-12628Altres ajuts: Applied Research Collaboration East Midlands (ARC EM)Altres ajuts: National Institute for Health Research (NIHR)Altres ajuts: Imperial Biomedical Research Centre (NIHR)The aims of our study was compare adherence measured by the medical possession ratio (MPR), time until discontinuation and describe adverse events after adding a DPP-4i, SGLT-2i, or sulfonylureas (SU) to metformin in a primary care population with insufficient glycemic control. We used routinely-collected health data from the SIDIAP database. The included subjects were matched by propensity score. The follow-up period was up to 24 months or premature discontinuation. The primary outcomes were the percentage of subjects with good adherence, treatment discontinuation and adverse events among treatment groups. The proportion of patients with good adherence (MPR> 0.8) after the addition of DPP-4i, SGLT-2i or SU was 53.6%, 68.7%, and 43.0%, respectively. SGLT-2i users were 1.7 times more likely to achieve good adherence compared with DPP-4i users (odds ratio [OR]:1.72, 98% confidence interval [CI]:1.51, 1.96), and 2.8 times more likely compared with SU users (OR: 0.35, 98% CI: 0.07, 0.29). The discontinuation hazard ratios were 1.43 (98%CI: 1.26; 1.62) and 1.60 (98%CI: 1.42; 1.81) times higher among SGLT-2i and SU users than DPP-4i users during the follow-up period. No differences were observed for adverse events among the treatment groups. In conclusion, in our real-world setting, the combination of SGLT-2i with metformin was associated with better adherence. The mean time until discontinuation was longer in the SGLT-2i group in comparison with the DPP-4i or SU groups

Efectividad del tratamiento con mufer preconcepcional en mujeres del área de salud del policlínico Gustavo Aldereguía, 2010-2011

Se realizó un estudio prospectivo-observacional, con el objetivo de determinar la efectividad del tratamiento con mufer preconcepcional, en mujeres atendidas en el área de salud del policlínico docente universitario Dr. Gustavo Aldereguía, durante los años 2010 y 2011; debido a que en la actualidad la profilaxis de la anemia desde el estado pre-gravídico es una prioridad para garantizar la adecuada salud materna-perinatal. El universo estuvo constituido por las 1296 féminas seguidas desde el riesgo pre-concepcional con el tratamiento con mufer, que planificaron embarazarse y se escogió una muestra aleatoria simple de 816 mujeres que se embarazaron en el último trimestre del año 2010 y dieron su consentimiento. Los datos obtenidos de formularios y la revisión de carné obstétrico reflejaron las variables: edad materna, captación, paridad, estado nutricional, enfermedades asociadas al embarazo, perfiles hematológicos, edad gestacional al parto y complicaciones post-parto presentadas. Predominó la edad entre los 20 y 35 años, la captación precoz en la totalidad de los casos, las nulíparas (74,8%) y las normopeso (57,2%); no presentaron enfermedades asociadas el 56,8%. El perfil hematológico se comportó: en la captación, entre 12,6-13,9 g/L el 86,2%, en el segundo trimestre el 76,5% entre 12-12.6g/L y en el tercer trimestre el 67,0% entre 11-11,9 g/L. La edad gestacional predominante al parto fue entre 37 y 42 semanas (83,4%) y no presentaron complicaciones post parto el 79,1%. El adecuado seguimiento de las mujeres en el riesgo preconcepcional con el suplemento nutricional con mufer repercutió favorablemente en la prevención de la anemia gestaciona

Fermented wheat germ extract inhibits glycolysis/pentoses cycle enzymes and induces apoptosis through poly(ADP-ribose) polymerase activation in Jurkat T cell leukemia tumor cells

The fermented extract of wheat germ, trade name Avemar, is a complex mixture of biologically active molecules with potent anti-metastatic activities in various human malignancies. Here we report the effect of Avemar on Jurkat leukemia cell viability, proliferation, cell cycle distribution, apoptosis, and the activity of key glycolytic/pentose cycle enzymes that control carbon flow for nucleic acid synthesis. The cytotoxic IC(50) concentration of Avemar for Jurkat tumor cells is 0.2 mg/ml, and increasing doses of the crude powder inhibit Jurkat cell proliferation in a dose-dependent fashion. At concentrations higher than 0.2 mg/ml, Avemar inhibits cell growth by more than 50% (72 h of incubation), which is preceded by the appearance of a sub-G(1) peak on flow histograms at 48 h. Laser scanning cytometry of propidium iodide- and annexin V-stained cells indicated that the growth-inhibiting effect of Avemar was consistent with a strong induction of apoptosis. Inhibition by benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone of apoptosis but increased proteolysis of poly(ADP-ribose) indicate caspases mediate the cellular effects of Avemar. Activities of glucose-6-phosphate dehydrogenase and transketolase were inhibited in a dose-dependent fashion, which correlated with decreased (13)C incorporation and pentose cycle substrate flow into RNA ribose. This decrease in pentose cycle enzyme activities and carbon flow toward nucleic acid precursor synthesis provide the mechanistic understanding of the cell growth-controlling and apoptosis-inducing effects of fermented wheat germ. Avemar exhibits about a 50-fold higher IC(50) (10.02 mg/ml) for peripheral blood lymphocytes to induce a biological response, which provides the broad therapeutic window for this supplemental cancer treatment modality with no toxic effects

MARTX Toxin in the Zoonotic Serovar of Vibrio vulnificus Triggers an Early Cytokine Storm in Mice

Vibrio vulnificus biotype 2-serovar E is a zoonotic clonal complex that can cause death by sepsis in humans and fish. Unlike other biotypes, Bt2 produces a unique type of MARTXVv (Multifunctional-Autoprocessive-Repeats-in-Toxin; RtxA13), which is encoded by a gene duplicated in the pVvBt2 plasmid and chromosome II. In this work, we analyzed the activity of this toxin and its role in human sepsis by performing in vitro, ex vivo, and in vivo assays. First, we demonstrated that the ACD domain, present exclusively in this toxin variant, effectively has an actin-cross-linking activity. Second, we determined that the whole toxin caused death of human endotheliocytes and monocytes by lysis and apoptosis, respectively. Finally, we tested the hypothesis that RtxA13 contributes to human death caused by this zoonotic serovar by triggering an early cytokine storm in blood. To this end, we used a Bt2-SerE strain (R99) together with its rtxA13 deficient mutant, and a Bt1 strain (YJ016) producing RtxA11 (the most studied MARTXVv) together with its rtxA11 deficient mutant, as controls. Our results showed that RtxA13 was essential for virulence, as R99ΔΔrtxA13 was completely avirulent in our murine model of infection, and that R99, but not strain YJ016, induced an early, strong and dysregulated immune response involving the up-regulation of a high number of genes. This dysregulated immune response was directly linked to RtxA13. Based on these results and those obtained ex vivo (human blood), we propose a model of infection for the zoonotic serovar of V. vulnificus, in which RtxA13 would act as a sepsis-inducing toxin

Intervención educativa para prevenir la Anemia Ferropénica gestacional

Introducción: la anemia ferropénica gestacional se asocia a efectos indeseables maternos-perinatales.Objetivo: describir los resultados obtenidos en una intervención educativa sobre la anemia ferropénica en un grupo de gestantes pertenecientes al Policlínico Guillermo Tejas, en la provincia Las Tunas, durante los meses marzo a mayo del 2020. Métodos: se realizó un estudio cuasiexperimental. De un universo de 86 embarazadas del Grupo Básico de Trabajo #1, se escogió una muestra aleatoria simple de 37 gestantes del primer trimestre del embarazo con su consentimiento, sin diagnóstico de anemia. Los datos obtenidos de encuestas, formularios, test evaluativos validados por expertos reflejaron las variables necesarias para el estudio. Se realizaron 3 etapas: Diagnóstica se identificaron los factores de riesgo y necesidades de aprendizaje, intervención se aplicó un programa educativo utilizando técnicas participativas y evaluación se midió el nivel cognoscitivo alcanzado sobre la prevención de la anemia y control nutricional.Resultados: incidió la edad materna entre 20-34 años 27(73%), el preuniversitario terminado 18(48,6%), trabajaban 21(56,8%), el periodo intergenésico más de 2 años 21(56,8%), 23(62,2%) fueron normopeso, no consumían el suplemento prenatal adecuadamente 24(64,8%), no fue adecuado el patrón nutricional 22(59,4%) y el nivel de conocimientos era bajo 26(70.3%). Después de intervenir 33(89,9%) fue alto el nivel cognoscitivo y aceptable el control nutricional 32(86,5%).Conclusiones: con la intervención realizada se logró alto nivel cognoscitivo sobre la prevención de la anemia ferropénica gestacional evaluándose de aceptable el control nutricional con adecuada ganancia de peso materno y perfiles hematológicos en límites normales