Constraints to virus infection in Nicotiana benthamiana plants transformed with a potyvirus amplicon

<p>Abstract</p> <p>Background</p> <p>Plant genomes have been transformed with full-length cDNA copies of viral genomes, giving rise to what has been called 'amplicon' systems, trying to combine the genetic stability of transgenic plants with the elevated replication rate of plant viruses. However, amplicons' performance has been very variable regardless of the virus on which they are based. This has boosted further interest in understanding the underlying mechanisms that cause this behavior differences, and in developing strategies to control amplicon expression.</p> <p>Results</p> <p><it>Nicotiana benthamiana </it>plants were transformed with an amplicon consisting of a full-length cDNA of the potyvirus <it>Plum pox virus </it>(PPV) genome modified to include a GFP reporter gene. Amplicon expression exhibited a great variability among different transgenic lines and even among different plants of the same line. Plants of the line 10.6 initially developed without signs of amplicon expression, but at different times some of them started to display sporadic infection foci in leaves approaching maturity. The infection progressed systemically, but at later times the infected plants recovered and returned to an amplicon-inactive state. The failure to detect virus-specific siRNAs in 10.6 plants before amplicon induction and after recovery suggested that a strong amplicon-specific RNA silencing is not established in these plants. However, the coexpression of extra viral silencing suppressors caused some amplicon activation, suggesting that a low level of RNA silencing could be contributing to maintain amplicon repression in the 10.6 plants. The resistance mechanisms that prevent amplicon-derived virus infection were also active against exogenous PPV introduced by mechanical inoculation or grafting, but did not affect other viruses. Amplicon-derived PPV was able to spread into wild type scions grafted in 10.6 rootstocks that did not display signs of amplicon expression, suggesting that resistance has little effect on virus movement.</p> <p>Conclusions</p> <p>Our results suggest that amplicon-derived virus infection is limited in this particular transgenic line by a combination of factors, including the presumed low efficiency of the conversion from the transgene transcript to replicable viral RNA, and also by the activation of RNA silencing and other defensive responses of the plant, which are not completely neutralized by viral suppressors.</p

Argan pulp as a novel functional ingredient with beneficial effects on multiple metabolism biomarkers

Argan tree is mainly used to obtain the oil from its seeds in a process that generates by-products such as the pulp of the fruit. The aim of this study was to assess the in vitro antioxidant activity and profile of bioactive compounds from an argan fruit extract, and to assay the metabolic effects of the argan pulp intake in a mouse experimental model. Our results showed a high in vitro antioxidant activity in ethanolic extracts of argan juice and pulp measured as total polyphenol content, reducing capacity, ABTS, DPPH, and lipid peroxidation inhibition capacity. In addition, several bioactive compounds with antioxidant, hypoglycemic, and anti-inflammatory activity were identified (vaccihein A, bergapten, methyl chloragenate, 2-O-caffeoyl glucaric acid, avicularin, quercetin-3- O-sophoroside, prodelphinidin B4 and mirificin). Dietary intake of argan pulp induced a remarkable hypoglycemic activity and decreased the gene expression of inflammatory markers. Gene expression and enzyme activity of liver antioxidant and detoxifying enzymes were differentially affected by argan pulp intake, whereas gut microbiota was modulated decreasing the Firmicutes/Bacteroidetes ratio. In conclusion, argan pulp can be considered a functional food or ingredient with beneficial effects on several metabolism biomarkers.University of Granada, Junta de Andalucía, Spanish Ministry of Science, Innovation, and UniversitiesEuropean Union through projects PTQ-17-09172RTI2018- 100934-B-I00RTC2019-006870-1B-AGR-662-UGR20FEDER programAGR145CTS164CTS107 (Andalusian Government

Performance of Leishmania PFR1 recombinant antigen in serological diagnosis of asymptomatic canine leishmaniosis by ELISA

Abstract Background Leishmania infantum is a protozoan parasite transmitted by phlebotomine sand flies that causes life-threatening disease in humans and dogs. The dog is the primary reservoir of the parasite and early diagnosis of canine leishmaniosis is crucial at the clinical and epidemiological level. The currently available serological tests for CanL diagnostic show limitations therefore the aim of the present study was to investigate the diagnostic performance of an indirect antibody ELISA based on the Leishmania infantum recombinant antigen PFR1 in asymptomatically infected dogs. One hundred fifty-six dogs including Leishmania-free experimental Beagles and pet dogs from England, Scotland and Leishmania-endemic Murcia in Spain, were tested with the assay. The later were also tested with two commercial L. infantum crude antigen ELISAs (INgezim and Civtest, respectively) and a real-time kinetoplast PCR test. Results Anti-PFR1 antibodies were detected in the four groups of dogs, and the mean log-transformed optical density (OD) values were lowest in Beagles and in dogs from England and highest among dogs from Murcia (p < 0.05). Using the highest OD in beagles as the PFR1 ELISA cut-off point, the estimated seroprevalence was 27% (14-40%) in dogs from Murcia, 4% (0-9%) in dogs from Scotland and 3% (0-8%) in dogs from England (p < 0.05). Seroprevalence in dogs from Murcia according to the INgezim and Civtest ELISAs were 24% (12-37%) and 31% (18-45%), respectively, whilst the prevalence of infection based on PCR in these dogs was 73% (60-86). The percentages of PFR1-positive dogs that tested negative on the INgezim and Civtest ELISAs were 30% and 35%, respectively, and all of them tested positive on the PCR test. Relative to the PCR, the specificity, sensitivity and area under the ROC curve of the PFR1 ELISA were 100%, 36% and 0.74 (0.63-0.86), respectively. Conclusions The ability shown by the PFR1 ELISA to detect infected dogs that go undetected by the crude antigen ELISAs is clinically and epidemiologically useful and PFR1 could be considered a candidate for a multi-antigen-based immunoassay for early detection of L. infantum infected dogs

Temporal Integrative Analysis of mRNA and microRNAs Expression Profiles and Epigenetic Alterations in Female SAMP8, a Model of Age-Related Cognitive Decline

A growing body of research shows that epigenetic mechanisms are critically involved in normal and pathological aging. The Senescence-Accelerated Mouse Prone 8 (SAMP8) can be considered a useful tool to better understand the dynamics of the global epigenetic landscape during the aging process since its phenotype is not fully explained by genetic factors. Here we investigated dysfunctional age-related transcriptional profiles and epigenetic programming enzymes in the hippocampus of 2- and 9-month-old SAMP8 female mice using the Senescent-Accelerated Resistant 1 (SAMR1) mouse strain as control. SAMP8 mice presented 1,062 genes dysregulated at 2 months of age, and 1,033 genes at 9 months, with 92 genes concurrently dysregulated at both ages compared to age-matched SAMR1. SAMP8 mice showed a significant decrease in global DNA methylation (5-mC) at 2 months while hydroxymethylation (5-hmC) levels were increased in SAMP8 mice at 2 and 9 months of age compared to SAMR1. These changes were accompanied by changes in the expression of several enzymes that regulate 5-mC and methylcytosine oxidation. Acetylated H3 and H4 histone levels were significantly diminished in SAMP8 mice at 2-month-old compared to SAMR1 and altered Histone DeACetylase (HDACs) profiles were detected in both young and old SAMP8 mice. We analyzed 84 different mouse miRNAs known to be altered in neurological diseases or involved in neuronal development. Compared with SAMR1, SAMP8 mice showed 28 and 17 miRNAs differentially expressed at 2 and 9 months of age, respectively; 6 of these miRNAs overlapped at both ages. We used several bioinformatic approaches to integrate our data in mRNA:miRNA regulatory networks and functional predictions for young and aged animals. In sum, our study reveals interplay between epigenetic mechanisms and gene networks that seems to be relevant for the progression toward a pathological aging and provides several potential markers and therapeutic candidates for Alzheimer's Disease (AD) and age-related cognitive impairment

Development of a Novel NGS Methodology for Ultrasensitive Circulating Tumor DNA Detection as a Tool for Early-Stage Breast Cancer Diagnosis

Breast cancer (BC) is the most prevalent cancer in women. While usually detected when localized, invasive procedures are still required for diagnosis. Herein, we developed a novel ultrasensitive pipeline to detect circulating tumor DNA (ctDNA) in a series of 75 plasma samples from localized BC patients prior to any medical intervention. We first performed a tumor-informed analysis to correlate the mutations found in tumor tissue and plasma. Disregarding the tumor data next, we developed an approach to detect tumor mutations in plasma. We observed a mutation concordance between the tumor and plasma of 29.50% with a sensitivity down to 0.03% in mutant variant allele frequency (VAF). We detected mutations in 33.78% of the samples, identifying eight patients with plasma-only mutations. Altogether, we determined a specificity of 86.36% and a positive predictive value of 88.46% for BC detection. We demonstrated an association between higher ctDNA median VAF and higher tumor grade, multiple plasma mutations with a likelihood of relapse and more frequent TP53 plasma mutations in hormone receptor-negative tumors. Overall, we have developed a unique ultra-sensitive sequencing workflow with a technology not previously employed in early BC, paving the way for its application in BC screening.Comino-Mendez’s contract is funded by the Spanish Association Against Cancer Scientific Foundation (AECC). This study was supported by the “Consejería de Salud y Familias—Junta de Andalucía” (PI-0291-2019), “Fundación Unicaja” is funding Alba-Bernal’s contract and the Andalusia-Roche Network in Precision Medical Oncology Quirós-Ortega’s contract. Carbajosa-Antona’s contract is funded by the “Ayudas María Zambrano para la atracción de talento internacional—Universidad de Málaga”. Partial funding for open access charge: Universidad de Málag

Effect of high-volume insecticide spraying on sand fly vectors in household gardens in Spain

This study investigated the efficacy of high-volume spraying with the adulticide α-cypermethrin alone and in combination with the larvicide diflubenzuron on the density of sand flies in gardens of three detached households in periurban areas in southeast Spain. Treatments were applied four times between June and August 2016, and four nearby sites, two households and two non-urbanized sites, were untreated controls. The number of sand flies collected between May and October 2016 using sticky interception and light attraction traps, was 4446 specimens. Species identified morphologically included Sergentomyia minuta (n = 2101; 48%), Phlebotomus perniciosus (n = 1922; 44%), Phlebotomus papatasi (n = 173; 4%), Phlebotomus sergenti (n = 161; 4%) and Phlebotomus ariasi (n = 36; 1%). Sand flies were detected in both treated and untreated sites. The proportion of positive sticky traps and the median (range) density of sand flies in positive traps were 61% traps and 7 (2-172) sand flies/m2 /day in untreated sites, and 43% traps and 4 (1-56) sand flies/m2 /day in treated sites (p < 0.05). Similarly, for light traps, it was 96% traps and 30 (3-168) flies/trap/day, and 83% traps and 3 (1-12) sand flies/trap/day, respectively (p < 0.05). However, sand fly density followed a comparable seasonal pattern in untreated and treated sites and did not consistently decrease following insecticide applications. These results were confirmed with mixed negative binomial modelling of sand fly density adjusted for time since application, month, environmental setting and site. The limited efficacy of the treatments, added to their cost, the impact of insecticides on non-target organisms and human health, and the risk of development of insecticide resistance, should dissuade similar outdoor applications to control sand fly vector populations in residential areas.Open Access funding was provided, thanks to the CRUE-CSIC UM agreement with Wiley.The study was funded by the Spanish Ministry of Science and Innovation (Grant number: AGL2013-46981-R) and received support from VectorNet, a European network for sharing data on the geographic distribution of arthropod vectors, transmitting human and animal disease agents funded by the European Food Safety Authority (EFSA) and the European Centre for Disease Prevention and Control (ECDC). CMH held a PhD grant from the University of Murcia (Contrato predoctoral FPU). EB, MO, CMH, JR and SV were beneficiaries of a COST TD1303 short scientific mission grant to develop sand fly sampling and identification techniques. CMH is now supported by Margarita Salas Postdoctoral Grant in University of Murcia from the Program of Requalification of the Spanish University System (Spanish Ministry of Universities), founded by the European Union—NextGenerationEU.Peer reviewe

New evidences of historical surface rupture on the Alhama de Murcia fault: implications for seismic hazard

Reunión Ibérica sobre Fallas Activas y Paleosismología (3ª. 2018. Alicante). - Texto en inglés con resumen y palabras claves en inglés y españolThe date of the last large event occurred in a fault segment is of great importance in seismic hazard assessment, especially when time-dependent models are considered. Eight of the twelve destructive earthquakes occurred in the eastern Betic Cordillera since 16th century, are located less than 10 km away from the Alhama de Murcia fault (AMF). Up to now, ground surface geological evidence to associate these events with the activity of specific fault segments has not been identified. In this work we present the first geological evidence of the catastrophic 1674 event occurred at Lorca (SE Spain). The excavations at La Torrecilla creek exposed archaeological remains from the Islamic period affected by 55+/-20 cm offset by the AMF fault. This supports the occurrence of earthquakes with surface rupture in the historical period on the Alhama de Murcia Fault and reinforces the results obtained in the previous paleoseismological works. The theoretical scenarios of maximum magnitudes and recurrence time obtained by combining this historical event with the fault slip rate, given the accumulated slip deficit, support that the seismic hazard associated with maximum magnitude events in this section of AMF could be greater than previously thought = La edad del último gran terremoto en un segmento de una falla sismogénica es un parámetro clave para los cálculos de peligrosidad sísmica, especialmente en los cálculos basados en modelos dependientes del tiempo. Ocho de los doce terremotos destructivos ocurridos en las cordilleras Béticas orientales desde el siglo XVI están localizados a menos de 10 km de la traza superficial de la Falla de Alhama de Murcia (AMF). Hasta ahora no se han identificado evidencias geológicas de ruptura superficial asociada a estos eventos que permitan asociarlos a segmentos específicos de la AMF. En este trabajo se presentan evidencias geológicas de ruptura superficial asociada al terremoto de Lorca de 1674. Las excavaciones paleosísmicas realizadas en la rambla de La Torrecilla han expuesto restos de una acequia datada como del periodo árabe que aparece afectada por un desplazamiento de 55+/-20 cm de la falla AMF. Esto apoya la ocurrencia de eventos con ruptura superficial en época histórica a lo largo de la AMF y refuerza las observaciones obtenidas en estudios paleosísmicos previos. Los escenarios teóricos de eventos de magnitud máxima e intervalos de recurrencia, combinando este evento histórico con los valores de tasa de desplazamiento de la AMF disponibles, considerando el déficit de desplazamiento acumulado, apoyan que la peligrosidad sísmica asociada a eventos de magnitud máxima en esta sección de la AMF puede ser mayor de la considerada hasta ahora.Departamento de Geodinámica, Estratigrafía y Paleontología, Universidad Complutense de Madrid, EspañaInstituto de Geociencias, Consejo Superior de Investigaciones Científicas, EspañaInstituto de Geociencias, Universidad Complutense de Madrid, EspañaDepartamento de Geología y Geoquímica, Universidad Autónoma de Madrid, EspañaInstituto Geológico y Minero de España, EspañaDepartamento de Ingeniería Topográfica y Cartografía, Universidad Politécnica de Madrid, EspañaGrupo RISKNAT, Departament de Dinàmica de la Terra i de l’Oceà, Universitat de Barcelona, EspañaAIA Estudio, Españ

Introducción a la química industrial

Fac. de Ciencias QuímicasTRUEpu

Towards Sustainable Innovative Business Models

This paper addresses two research questions: (1) How do firms innovate their business models to deal with the economic, environmental and social aspects of sustainability, and their interconnections? (2) How do managers design the process toward more sustainable innovative business models? Starting from the triple-layered canvas as a theoretical framework, a pattern matching technique is used to compare that theoretical pattern to the empirical pattern observed. The study is based on qualitative methods and data from a Spanish company in the wine sector. The findings indicate that the triple-layered canvas is applicable and useful for micro firms, although the process to design sustainable innovative business models might be even more important. A new theoretical model is inferred and proposed to incorporate the perspective of the process of the business model innovations for sustainability, and to add several relevant aspects to make the process more successful. Besides this, non-family firms introducing sustainable business model innovations in their economic, ecological, and social aspects move closer to family firms’ distinctive behavior. Finally, the implications and future lines of research are summarized

Supramolecular Coordination of Pb2+ Defects in Hybrid Lead Halide Perovskite Films Using Truxene Derivatives as Lewis Base Interlayers

Truxene derivatives, due to their molecular structure and properties, are good candidates for the passivation of defects when deposited onto hybrid lead halide perovskite thin films. Moreover, their semiconductor characteristics can be tailored through the modification of their chemical structure, which allows‐upon light irradiation‐ the interfacial charge transfer between the perovskite film and the truxene molecules. In this work, we analysed the use of the molecules as surface passivation agents and their use in complete functional solar cells. We observed that these molecules reduce the non‐radiative carrier recombination dynamics in the perovskite thin film through the supramolecular complex formation between the Truxene molecule and the Pb2+ defects at the perovskite surface. Interestingly, this supramolecular complexation neither affect the carrier recombination kinetics nor the carriers collection but induced noticeable hysteresis on the photocurrent vs voltage curves of the solar cells under 1 sun illumination