Video design adaptation to youtube advertising formats

Este trabajo en curso analiza cómo diferentes diseños publicitarios en términos de arousal (intensidad emocional) pueden contribuir a mejorar la efectividad de la video publicidad online. La literatura previa sugiere que un final con alto arousal puede incrementar la efectividad publicitaria. Para comprobar esta proposición, nuestra investigación combina diferentes metodologías: neurociencia aplicada al comportamiento del consumidor; un estudio de campo basado en una campaña publicitaria en YouTube y un estudio final. Los primeros resultados sugieren que el diseño publicitario ha de adaptarse a los nuevos formatos de video publicidad interactiva en función de los objetivos de los anunciantes.This work in progress analyzes how different arousal advertising design may enhance online video advertising effectiveness. Previous research suggests that arousing ending designs could increase ad effectiveness. To test this proposition, our research combines different methodologies: a consumer neuroscience pretest is used to identify high and low arousal sequences; a field study based on a YouTube ad campaign is used as an exploratory study which helps to determine the experimental scenarios to be used in the final study with a large sample size. Preliminary results suggest that ad design need to be adapted to new formats of interactive advertising depending on the advertisers’ goals

A bacterial acetyltransferase targets the protein kinase ZIP1, a positive regulator of plant immunity

Pseudomonas syringae is a model bacterial pathogen that penetrates the leaf to reach the plant apoplast, where it replicates causing disease. In order to do that, the pathogen must interfere and suppress a two-tiered plant defense response: PTI (PAMP-Triggered Immunity, or basal resistance) and ETI (Effector-Triggered Immunity). P. syringae uses a type III secretion system to directly deliver effector proteins inside the plant cell cytosol, many of which are known to suppress PTI, some of which are known to trigger ETI, and a handful of which are known to suppress ETI. Bacterial infection can also trigger a systemic plant defense response that protects the plant against additional pathogen attacks known as SAR (Systemic Acquired Resistance). We are particularly interested in the molecular and cellular mechanisms involved in effector-mediated defense evasion by P. syringae, in particular those involved in the suppression of ETI and SAR, and/or mediation of hormone signaling. Here we present data describing effector-mediated interference with plant immunity, by means of acetylation of a key positive regulator of local and systemic responses. Our work identifies a novel plant target for effector function, and characterizes its function. This work illustrates how analyzing the means by which a given effector interferes with its target can provide novel information regarding eukaryotic molecular mechanisms.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. MINECO BIO2015-64391R y FEDE

The bacterial effector HopZ1a acetylates ZIP1 kinase to suppress Arabidopsis defence responses

During the plant-pathogen interaction, disease or resistance are determined in the plant by a series of molecular events. The plant detects Pathogen-Associated Molecular Patterns (PAMPs), such as flagellin, triggering a defence response called PTI (PAMP-Triggered Immunity). Bacterial pathogens can in turn suppress such defence response through the translocation into the plant cell cytosol of virulence proteins, called effectors, via a Type Three Secretion System (T3SS). In resistant plants, intracellular receptors known as R proteins recognize these effectors, triggering a second line of defence, more specific and intense, called ETI (Effector-Triggered Immunity), which usually leads to programmed cell death known as HR (Hypersensitive Response). Pseudomonas syringae is a phytopathogenic bacterium whose virulence depends on a T3SS and its effector repertoire. Some strains include HopZ1a, an unusual effector which is able to suppress in Arabidopsis both local (PTI and ETI), and systemic (SAR, for Systemic Acquired Resistance) defences, by means of its acetyltransferase activity. In resistant Arabidopsis plants, HopZ1a acetylates the ZED1 pseudokinase, which is proposed to function as a decoy mimicking HopZ1a target in the plant: ZED1 modification activates an R-protein (ZAR1) to trigger HopZ1a-dependent ETI. None of the Arabidopsis proteins proposed to date as HopZ1a targets is a kinase, nor fully explains the effector´s defence suppression abilities. In this work we identify an Arabidopsis kinase that functions as a positive regulator of PTI, ETI and SAR, which interacts with HopZ1a and is acetylated by this effector in lysine residues essential for its kinase activity. Further, HopZ1a can specifically suppress the defence phenotypes resulting from ZIP1 expression in Arabidopsis. We propose that ZIP1 acetylation by HopZ1a interferes with its kinase activity, and consequently with positive defence signalling.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

The bacterial effector HopAF1 interacts with Arabidopsis MKK4/5

Pseudomonas syringae is a phytopathogenic bacterium whose virulence depends on a Type III Secretion System and its effector (T3E) repertoire, which is translocated into the host cell cytosol suppressing both basal and effector-triggered Immunity (ETI). HopAF1 is a T3E that suppresses several plant immunity phenotypes, including flg22-induced ethylene (ET) production. HopAF1 is mainly located in the plasma membrane (PM), where it interacts with plant proteins MTN1/2 that participate in ET biosynthesis. However, PM localization is not essential for HopAF1 ability to repress ET accumulation. While seeking for additional HopAF1 interactors in the plant, we have identified MAP kinase kinases MKK4/5. The MAP kinase module including MKK4/5 and MPK3/6 positively regulates 1-aminocyclopropane-1-carboxylic acid synthase (ACS) activity, a rate-limiting and major regulatory step in stress-induced ET production. MKK4/5-dependent phosphorylation directly stabilizes ACS2/6 and indirectly induces ACS transcription. We originally identified HopAF1-MKK4 interaction through an MS-based screening, and have now confirmed it by pull-down and BIFC, while obtaining variable results using FRET-FLIM. Moreover, we are monitoring ACS2/6 expression levels by qRT-PCR, in the presence and absence of HopAF1. We propose that HopAF1 interference with MKK4/5 is likely to contribute to the suppression of ET production, by altering MKK4/5-dependent regulation of ACS activity.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

El efector bacteriano HopZ1a acetila ZIP1 para suprimir las respuestas de defensa en Arabidopsis

Pseudomonas syringae es un patógeno bacteriano modelo capaz de infectar y desarrollar la enfermedad en una gran variedad de plantas. Para ello, P. syringae utiliza un sistema de secreción de tipo III para translocar proteínas efectoras dentro del citosol de la célula vegetal, muchas de las cuales suprimen la defensa basal PTI (PAMP-Triggered Immunity), disparada por el reconocimiento de patrones moleculares asociados a patógenos (PAMPs). Algunos de esos efectores suprimen la ETI (Effector-Triggered Immunity), respuesta de defensa más específica e intensa desencadenada por el reconocimiento de efectores y cuyo resultado final es la muerte celular programada llamada HR (Hypersensitive Response). Además de las respuestas locales, PTI y ETI, la planta puede activar una respuesta sistémica de defensa SAR (Systemic Acquire Resistance) que la protege contra ataques posteriores del patógeno. El efector HopZ1a es una acetiltransferasa perteneciente a la superfamilia de efectores YopJ, capaz de suprimir las respuestas de defensa PTI, ETI y SAR en Arabidopsis. De entre todas las diana descritas para este efector, ZED1 es una pseudokinasa cuya única función conocida es ser acetilada por HopZ1a, activando así a una proteína R llamada ZAR1, y desencadenando ETI. Esta función de decoy de ZED1, junto a la existencia de interacciones entre otros efectores de la familia YopJ con kinasas de animales, nos lleva a pensar que una kinasa pueda ser diana de HopZ1a. Nuestro trabajo identifica a la kinasa ZIP1, regulador positivo de defensa, como diana de HopZ1a. Nosotros caracterizamos su interacción y demostramos que HopZ1a acetila a ZIP1 en una lisina esencial para su función kinasa para suprimir las defensas de la planta.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

El regulador positivo de defensas ZIP1 es acetilado por el efector bacteriano HopZ1a para suprimir las respuestas de defensa en Arabidopsis

Durante la interacción planta-patógeno, el éxito en la colonización y desarrollo de la enfermedad del patógeno viene determinado por su capacidad de evadir y/o suprimir las respuestas de defensas disparadas por la planta. La planta detecta patrones moleculares, como por ejemplo flagelina o EF-Tu, desencadenando una defensa llamada PTI , que algunas bacteriaspatógenas pueden suprimir mediante la proteínas de virulencia (efectores) . En plantas resistentes, la célula detecta dichos efectores mediante receptores intracelulares, proteínas R, desencadenando una segunda línea de defensa, llamada ETI , que conduce a una muerte celular programada conocida como HR. La activación de estas respuestas de defensa a nivel local inducen la Respuesta Sistémica Adquirida (SAR) en hojas distales. Pseudomonas syringae es una bacteria fitopatógena, capaz de infectar una gran variedad de plantas, incluyendo Arabidopsis. Su virulencia depende de su capacidad de translocar efectores Tipo III (T3Es), directamente al citosol de la célula hospedadora mediante un Sistema de Secreción Tipo III (T3SS). HopZ1a es un efector capaz de suprimir en Arabidopsis defensas locales (PTI y ETI) y sistémicas (SAR) por medio de su actividad acetiltransferasa. En bacterias patógenas de animales, los homólogos de HopZ1a inactivan por acetilación a quinasas del hospedador implicadas en señalización positiva de defensa. En plantas resistentes, HopZ1a acetila la pseudoquinasa ZED1, propuesta como señuelo que imita al objetivo HopZ1a en la planta, para desencadenar la ETI dependiente de HopZ1a. Estos datos sugieren una quinasa reguladora positiva de defensa, tanto a nivel local como sistémico, como probable diana de HopZ1a. En este trabajo identificamos a ZIP1, una que funciona como un regulador positivo de PTI, ETI y SAR, que interactúa con HopZ1a y es acetilada por este efector en una lisina esencial para su actividad quinasa.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

The bacterial effector HopZ1a acetylates MKK7 to suppress plant immunity

The Pseudomonas syringae type III secretion system translocates effector proteins into the host cell cytosol to suppress plant basal immunity. Effector HopZ1a suppresses local and sys- temic immunity triggered by pathogen-associated molecular patterns (PAMPs) and effectors, through target acetylation. HopZ1a has been shown to target several plant proteins, but none fully substantiates HopZ1a-associated immune suppression. Here, we investigate Arabidopsis thaliana mitogen-activated protein kinase kinases (MKKs) as potential targets, focusing on AtMKK7, a positive regulator of local and systemic immunity. We analyse HopZ1a interference with AtMKK7 by translocation of HopZ1a from bacteria inoculated into Arabidopsis expressing MKK7 from an inducible promoter. Reciprocal pheno- types are analysed on plants expressing a construct quenching MKK7 native expression. We analyse HopZ1a–MKK7 interaction by three independent methods, and the relevance of acetylation by in vitro kinase and in planta functional assays. We demonstrate the AtMKK7 contribution to immune signalling showing MKK7- dependent flg22-induced reactive oxygen species (ROS) burst, MAP kinas (MAPK) activation and callose deposition, plus AvrRpt2-triggered MKK7-dependent signalling. Furthermore, we demonstrate HopZ1a suppression of all MKK7-dependent responses, HopZ1a–MKK7 interac- tion in planta and HopZ1a acetylation of MKK7 with a lysine required for full kinase activity. We demonstrate that HopZ1a targets AtMKK7 to suppress local and systemic plant immunity.RTI2018- 095069-B-I00, BIO2015-64391-R, UMA18-FEDERJA-070, FPU14/04233, XDB27040204, Universidad de Málaga-CBU

SCORE and REGICOR function charts underestimate the cardiovascular risk in Spanish patients with rheumatoid arthritis

Introduction: Our objective was to determine which one of the two function charts available in Spain to calculate cardiovascular (CV) risk, Systematic COronary Risk Evaluation (SCORE) or Framingham-REgistre GIroní del COR (REGICOR), should be used in patients with rheumatoid arthritis (RA). Methods: A series of RA patients seen over a one-year period without history of CV events were assessed. SCORE, REGICOR, modified (m)SCORE and mREGICOR according to the European League Against Rheumatism (EULAR) recommendations were applied. Carotid ultrasonography (US) was performed. Carotid intima-media thickness (cIMT) > 0.90 mm and/or carotid plaques were used as the gold standard test for severe subclinical atherosclerosis and high CV risk (US+). The area under the receiver operating curves (AUC) for the predicted risk for mSCORE and mREGICOR were calculated according to the presence of severe carotid US findings (US+). Results: We included 370 patients (80% women; mean age 58.9 ± 13.7 years); 36% had disease duration of 10 years or more; rheumatoid factor (RF) and/or anticyclic citrullinated peptide (anti-CCP) were positive in 68%; and 17% had extra-articular manifestations. The EULAR multiplier factor was used in 122 (33%) of the patients. The mSCORE was 2.16 ± 2.49% and the mREGICOR 4.36 ± 3.46%. Regarding US results, 196 (53%) patients were US+. The AUC mSCORE was 0.798 (CI 95%: 0.752 to 0.844) and AUC mREGICOR 0.741 (95% CI; 0.691 to 0.792). However, mSCORE and mREGICOR failed to identify 88% and 91% of US+ patients. More than 50% of patients with mSCORE ≥1% or mREGICOR >1% were US+. Conclusions: Neither of these two function charts was useful in estimating CV risk in Spanish RA patients

Suppression of HopZ Effector-Triggered Plant Immunity in a Natural Pathosystem

Many type III-secreted effectors suppress plant defenses, but can also activate effector-triggered immunity (ETI) in resistant backgrounds. ETI suppression has been shown for a number of type III effectors (T3Es) and ETI-suppressing effectors are considered part of the arms race model for the co-evolution of bacterial virulence and plant defense. However, ETI suppression activities have been shown mostly between effectors not being naturally expressed within the same strain. Furthermore, evolution of effector families is rarely explained taking into account that selective pressure against ETI-triggering effectors may be compensated by ETI-suppressing effector(s) translocated by the same strain. The HopZ effector family is one of the most diverse, displaying a high rate of loss and gain of alleles, which reflects opposing selective pressures. HopZ effectors trigger defense responses in a variety of crops and some have been shown to suppress different plant defenses. Mutational changes in the sequence of ETI-triggering effectors have been proposed to result in the avoidance of detection by their respective hosts, in a process called pathoadaptation. We analyze how deleting or overexpressing HopZ1a and HopZ3 affects virulence of HopZ-encoding and non-encoding strains. We find that both effectors trigger immunity in their plant hosts only when delivered from heterologous strains, while immunity is suppressed when delivered from their native strains. We carried out screens aimed at identifying the determinant(s) suppressing HopZ1a-triggered and HopZ3-triggered immunity within their native strains, and identified several effectors displaying suppression of HopZ3-triggered immunity. We propose effector-mediated cross-suppression of ETI as an additional force driving evolution of the HopZ family

Clinical and molecular characterization of steatotic liver disease in the setting of immune-mediated inflammatory diseases

Background & Aims: Growing evidence suggests an increased prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) in the context of immune-mediated inflammatory diseases (IMIDs). We aimed to clinically and mechanistically characterize steatotic liver disease (SLD) in a prospective cohort of patients with IMID compared to controls. Methods: Cross-sectional, case-control study including a subset of patients with IMID. Controls from the general population were age-, sex-, type 2 diabetes-, and BMI-matched at a 1:2 ratio. SLD was established using controlled attenuation parameter. Liver biopsies were obtained when significant liver fibrosis was suspected. Total RNA was extracted from freshly frozen cases and analyzed by RNA-seq. Differential gene expression was performed with ‘limma-voom’. Gene set-enrichment analysis was per formed using the fgsea R package with a preranked “limma t-statistic” gene list. Results: A total of 1,456 patients with IMID and 2,945 controls were included. Advanced SLD (liver stiffness measurement ≥9.7 kPa) (13.46% vs. 3.79%; p <0.001) and advanced MASLD (12.8% vs. 2.8%; p <0.001) prevalence were significantly higher among patients with IMID than controls. In multivariate analysis, concomitant IMID was an independent, and the strongest, predictor of advanced SLD (adjusted odds ratio 3.318; 95% CI 2.225-4.947; p <0.001). Transcriptomic data was obtained in 109 patients and showed 87 significant genes differentially expressed between IMID-MASLD and control-MASLD. IMID-MASLD cases displayed an enriched expression of genes implicated in pro-tumoral activities or the control of the cell cycle concomitant with a negative expression of genes related to metabolism. Conclusions: The prevalence of advanced SLD and MASLD is disproportionately elevated in IMID cohorts. Our findings suggest that IMIDs may catalyze a distinct MASLD pathway, divergent from classical metabolic routes, highlighting the need for tailored clinical management strategies.Acknowledgments: This work was fully suported by grants from the Spanish Instituto de Salud Carlos III (ISCIII-FEDER). Grant numbers: PI18/01304, PI20/01279, and PI19/00204This work was fully suported by grants from the Spanish Instituto de Salud Carlos III (ISCIII-FEDER). Grant numbers: PI18/01304, PI20/01279, and PI19/00204