Histone Mutants Separate R Loop Formation from Genome Instability Induction

R loops have positive physiological roles, but they can also be deleterious by causing genome instability, and the mechanisms for this are unknown. Here we identified yeast histone H3 and H4 mutations that facilitate R loops but do not cause instability. R loops containing single-stranded DNA (ssDNA), versus RNA-DNA hybrids alone, were demonstrated using ssDNA-specific human AID and bisulfite. Notably, they are similar size regardless of whether or not they induce genome instability. Contrary to mutants causing R loop-mediated instability, these histone mutants do not accumulate H3 serine-10 phosphate (H3S10-P). We propose a two-step mechanism in which, first, an altered chromatin facilitates R loops, and second, chromatin is modified, including H3S10-P, as a requisite for compromising genome integrity. Consistently, these histone mutations suppress the high H3S10 phosphorylation and genomic instability of hpr1 and sen1 mutants. Therefore, contrary to what was previously believed, R loops do not cause genome instability by themselves.European Research Council ERC2014 AdG669898Ministerio de Economía y Competitividad BFU2013-42918-P, BFU2016-75058-

Filósofas: presencia y asusencia del pensamiento femenino en los estudios de Filosofía

Memoria ID-192. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2018-2019

Detection and Genetic Diversity of Porcine Coronavirus Involved in Diarrhea Outbreaks in Spain

[EN]Porcine enteric coronaviruses include some of the most relevant viral pathogens to the swine industry such as porcine epidemic diarrhea virus (PEDV) or porcine transmissible gastroenteritis virus (TGEV) as well as several recently identified virus such as swine enteric coronavirus (SeCoV), porcine deltacoronavirus (PDCoV) or swine enteric alphacoronavirus (SeACoV). The aim of this study is the identification and characterization of enteric coronaviruses on Spanish pig farms between 2017 and 2019. The study was carried out on 106 swine farms with diarrhea outbreaks where a viral etiology was suspected by using two duplex RT-PCRs developed for the detection of porcine enteric coronaviruses. PEDV was the only coronavirus detected in our research (38.7% positive outbreaks, 41 out of 106) and neither TGEV, SeCoV, PDCoV nor SeACoV were detected in any of the samples. The complete S-gene of all the PEDV isolates recovered were obtained and compared to PEDV and SeCoV sequences available in GenBank. The phylogenetic tree showed that only PEDV of the INDEL 2 or G1b genogroup has circulated in Spain between 2017 and 2019. Three different variants were detected, the recombinant PEDV-SeCoV being the most widespread. These results show that PEDV is a relevant cause of enteric disorders in pigs in Spain while new emerging coronavirus have not been detected so far. However, the monitoring of these virus is advisable to curtail their emergence and spread.SIThis work was supported by the program from the National Institute of Agricultural and Food Research and Technology (INIA project E-RTA2015-0003-C02-02) of Spanish Government. HP, ÓM-A, and HA were supported by Spanish Government (FPU17/00466, FPU16/03485, and BEAGAL-18-106, respectively) and MG-G by Junta de Castilla y León (LE131-18)

Feasibility and efficacy of a virtual reality game-based upper extremity motor function rehabilitation therapy in patients with chronic stroke: a pilot study

This article belongs to the Special Issue New Technologies in the Evaluation and Treatment of Patients with Neurological Pathologies.Background: The objective of the present study was to develop a virtual reality protocol based on activities of daily living and conventional rehabilitation, using Leap Motion Controller to improve motor function in upper extremity rehabilitation in stroke patients. At the same time, the purpose was to explore its efficacy in the recovery of upper extremity motor function in chronic stroke survivors, and to determine feasibility, satisfaction and attendance rate; Methods: A prospective pilot experimental clinical trial was conducted. The outcome measures used were the grip strength, the Action Research Arm Test (ARAT), the Block and Box Test (BBT), the Short Form Health Survey-36 Questionnaire, a satisfaction questionnaire and attendance rate; Results: Our results showed statistically significant changes in the variables grip strength, BBT and ARAT as well as high levels of satisfaction and attendance; Conclusions: This virtual reality platform represents an effective tool in aspects of upper extremity functionality rehabilitation in patients with chronic stroke, demonstrating feasibility and high levels of attendance and satisfaction

Effects of performing morning versus afternoon exercise on glycemic control and hypoglycemia frequency in type 1 diabetes patients on sensor-augmented insulin pump therapy

Q1Artículo original619-624Background: Although physical exercise (PE) is recommended for individuals with type 1 diabetes (DM1), participation in exercise is challenging because it increases the risk of severe hypoglycemia and the available therapeutic options to prevent it frequently result in hyperglycemia. There is no clear recommendation about the best timing for exercise. The aim of this study was to compare the risk of hypoglycemia after morning or afternoon exercise sessions up to 36 hours postworkout. Methods: This randomized crossover study enrolled subjects with DM1, older than 18 years of age, on sensor-augmented insulin pump (SAP) therapy. Participants underwent 2 moderate-intensity exercise sessions; 1 in the morning and 1 in the afternoon, separated by a 7 to 14 day wash-out period. Continuous glucose monitoring (CGM) data were collected 24 hours before, during and 36 hours after each session. Results: Thirty-five subjects (mean age 30.31 ± 12.66 years) participated in the study. The rate of hypoglycemia was significantly lower following morning versus afternoon exercise sessions (5.6 vs 10.7 events per patient, incidence rate ratio, 0.52; 95% CI, 0.43-0.63; P < .0001). Most hypoglycemic events occurred 15-24 hours after the session. On days following morning exercise sessions, there were 20% more CGM readings in near-euglycemic range (70-200 mg/dL) than on days prior to morning exercise (P = .003). Conclusions: Morning exercise confers a lower risk of late-onset hypoglycemia than afternoon exercise and improves metabolic control on the subsequent day

Análise bibliométrica de teses de doutorado espanholas em aleitamento materno, entre os años 1980-2015

La sociedad actual inserta en un mundo VUCA (volátil, incierto, complejo y ambiguo) ha producido cambios vertiginosos en relación a la Lactancia Materna (LM). Es por ello que consideramos preciso analizar el conocimiento y la producción científica efectuada hasta la fecha sobre esta disciplina. De esta inquietud, surge esta investigación como un estudio interdisciplinar que pretende dar prioridad a la protección, promoción y apoyo a la LM. La finalidad es investigar el crecimiento y la distribución de los documentos científicos a través de su análisis y evaluación, con el fin de recoger la producción científica de los investigadores en LM. Para ello se utiliza una muestra de 155 Tesis Doctorales publicadas en las Universidades Españolas, con el denominador común de la palabra clave lactancia materna, en el período comprendido entre los años 1980 a 2015. La metodología utilizada es mixta, con análisis de datos cualitativos, enmarcados dentro de un análisis retrospectivo y descriptivo, así como un análisis cuantitativo mediante indicadores bibliométricos. Se trata de ofrecer un conjunto sólido y fiable de procesos y resultados sistemáticos, empíricos y críticos de investigación, a fin de exponer la información sobre las tendencias actuales y futuras de investigación e innovación.The current society inserted in a VUCA world (volatile, uncertain, complex and ambiguous) has produced vertiginous changes in relation to breastfeeding. That is why we consider it necessary to analyze the knowledge and scientific production carried out to date on this discipline. From this concern, this research emerges as an interdisciplinary study that aims to give priority to the protection, promotion and support of breastfeeding. The purpose is to investigate the growth and distribution of scientific documents through their analysis and evaluation, in order to gather the scientific production of researchers on breastfeeding. To this end, a sample of 155 Doctoral Theses published in Spanish Universities is used, with the common denominator of the word breastfeeding, in the period that goes from 1980 to 2015. The methodology used is mixed, with qualitative data analysis, framed within a retrospective and descriptive analysis, as well as a quantitative analysis through bibliometric indicators. The aim is to offer a solid and reliable set of systematic, empirical and critical research processes and results, in order to present information on current and future research and innovation trends.A sociedade atual inserida em um mundo da VUCA (volátil, incerto, complexo e ambíguo) produziu mudanças vertiginosas em relação ao aleitamento materno. Por isso, consideramos necessário analisar o conhecimento e a produção científica realizada até o momento nesta disciplina. A partir desta preocupação, esta pesquisa surge como um estudo interdisciplinar que visa dar prioridade à proteção, promoção e apoio da aleitamento materno. Objetivo é investigar o crescimento e a distribuição de documentos científicos por meio de sua análise e avaliação, a fim de coletar a produção científica de pesquisadores em aleitamento materno. Para tanto, utiliza-se uma amostra de 155 teses de doutorado publicadas em universidades espanholas, com o denominador comum da palavra amamentação, no período de 1980 a 2015. A metodologia utilizada é mista, com análise qualitativa dos dados, enquadrado em uma análise retrospectiva e descritiva, bem como uma análise quantitativa através de indicadores bibliométricos. O objetivo é oferecer um conjunto sólido e confiável de processos e resultados sistemáticos, empíricos e críticos de pesquisa, a fim de apresentar informações sobre tendências atuais e futuras de pesquisa e inovação

Combined in-vitro and on-farm evaluation of commercial disinfectants used against Brachyspira hyodysenteriae

[EN] Background: Swine dysentery (SD) is a severe infectious disease with a relevant impact on pig production usually caused by Brachyspira hyodysenteriae, although B. hampsonii causes an identical clinical picture. SD control relies on antimicrobials, good management practices and strict biosecurity with cleaning and disinfection as crucial tools to avoid the pathogen transmission. This study evaluates the in-vitro efficacy of an array of commercial disinfectants against a collection of B. hyodysenteriae isolates using broth tests. The efficacy of cleaning and disinfection protocols was also evaluated on two farms with endemic SD using surface swabs collected in emptied pens before and after cleaning and disinfection procedures, using both real-time PCR and bacterial microbiological culture. Results: Most of the commercial disinfectants evaluated were effective against all B. hyodysenteriae isolates tested, with a reduction of more than 5.00 log10 CFU/mL (bactericidal efficacy of 99.999%). However, some isolates exhibited reduced susceptibility to Virkon-S and Limoseptic disinfectants. The evaluation of cleaning and disinfection protocols on farms with SD outbreaks showed that approximately half the pens tested (n = 25) were positive by real-time PCR after pigs removal (mean B. hyodysenteriae counts 5.72 ± 1.04 log10 CFU/mL) while almost 20% of the pens remained positive after cleaning (n = 7) and disinfection (n = 5) procedures although with significantly lower, mean estimates (4.31 ± 0.43 log10 CFU/mL and 4.01 ± 0.55 log10 CFU/mL, respectively). Conclusions: These results show the efficacy of disinfectants against B. hyodysenteriae but also stress the need to implement adequately the cleaning and disinfection protocols on pig farms and review and revise their efficiency periodically.SIThis study was supported by the Ministerio de Ciencia e Innovación (PID2019-110662RB-I00) and the Castilla y León Rural Development Pro‑ gramme co-fnanced by EAFRD (47/18/125/G0

In vitro evaluation of gentamicin activity against Spanish field isolates of Brachyspira hyodysenteriae

[EN]Background: The treatment of swine dysentery (SD) has become constrained in recent years due to the limited availability of effective drugs combined with a rise in antimicrobial resistance. Gentamicin, an aminoglycoside antibiotic, is authorised for the control of this disease in several European countries but has not been extensively used so far. In this study, the in vitro susceptibility of 56 Brachyspira hyodysenteriae field isolates was evaluated against gentamicin using a broth microdilution test. The molecular basis of decreased susceptibility to gentamicin was also investigated by sequencing the 16S rRNA gene and phylogenetic relatedness by multiple-locus variable number tandem-repeat analysis (MLVA). Results: Most B. hyodysenteriae isolates presented low minimum inhibitory concentration (MIC) values to gentamicin, with a mode of 2 µg/mL, a median or MIC50 of 4 µg/mL and percentile 90 or MIC90 of 16 µg/mL. The distribution of these values over the period studied (2011–2019) did not show a tendency towards the development of resistance to gentamicin. Differences in susceptibility among isolates could be explained by two point-mutations in the 16S rRNA gene, C990T and A1185G, which were only present in isolates with high MICs. These isolates were typed in three different MLVA clusters. Analyses of co-resistance between gentamicin and antimicrobials commonly used for the treatment of SD revealed that resistance to tiamulin and valnemulin was associated with low MICs for gentamicin. Conclusions: The results provide an accurate characterisation of antimicrobial sensitivity to gentamicin and possible mechanisms of resistance in Spanish B. hyodysenteriae isolates. These findings allow us to propose gentamicin as an alternative in the antibiotic management of SD, particularly in outbreaks caused by pleuromutilin resistant isolates.SIThe authors wish to thank Benjamín Rabanal for his contribution in some parts of the research and Diana Molina for her excellent technical assistance. Clara Vega (workplace ULE-02-B) and Manuel Gómez (LE131-18) are supported by grants from Junta de Castilla y León co-financed by the European Social Fund and Youth Employment Initiative. Héctor Argüello is supported by a grant from the Spanish Ministry of Education under the programme “Beatriz Galindo” (BEAGAL-18-106). Lucía Pérez-Pérez (PRE2020-093762) and Héctor Puente (FPU17/00466) are also supported by the Spanish Ministry of Education. We fnally want to thank the reviewer for the advice and ideas provided which clearly improved the quality of the manuscript.This work was supported by Fatro Ibérica S.L

In-depth in vitro Evaluation of the Activity and Mechanisms of Action of Organic Acids and Essential Oils Against Swine Enteropathogenic Bacteria

P. 1-13Alternative antimicrobials require a deep understanding of their action mechanisms by in vitro assays which support science-based field use. This study focuses on the characterization of bactericidal mechanisms of potential antimicrobial compounds, two organic acids and three single essential oil (EO) compounds against swine enteropathogenic bacteria Escherichia coli, Salmonella enterica subsp. enterica serovar Typhimurium, and Clostridium perfringens. Target concentrations of the compounds were evaluated using the inhibitory potential of the vapor phase and bacterial viability after short-term exposure, while cell targets were disclosed using flow cytometry (FC), Fourier-transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM). All tested compounds exhibited vapor phase activity against the three bacterial species, except sodium salt of coconut fatty acid distillates against C. perfringens. Survival test results evidenced that effects on bacterial viability were concentration dependent and higher in single EO compounds than in organic acids. In detail, thymol and its isomer carvacrol were the most effective compounds. Further characterization of thymol and cinnamaldehyde activity revealed that thymol main target was the cell membrane, since it caused striking damages in the membrane permeability, integrity and composition evidenced by FC and FTIR in the three enteric pathogens. In contrast, cinnamaldehyde was more effective against enterobacteria than against C. perfringens and only caused slightly damages at the highest concentration tested. Its target at the molecular level differed between enterobacteria and C. perfringens isolates. The SEM micrographs allowed us to confirm the results previously obtained for both EO compounds by other techniques. Altogether, the study showed the straight effect of these antimicrobials, which could constitute relevant information to optimize their feed inclusion rates in field studies or field use.S

Antimicrobial use and production system shape the fecal, environmental, and slurry resistomes of pig farms

P. 1-17Background: The global threat of antimicrobial resistance (AMR) is a One Health problem impacted by antimicrobial use (AMU) for human and livestock applications. Extensive Iberian swine production is based on a more sustainable and eco-friendly management system, providing an excellent opportunity to evaluate how sustained differences in AMU impact the resistome, not only in the animals but also on the farm environment. Here, we evaluate the resistome footprint of an extensive pig farming system, maintained for decades, as compared to that of industrialized intensive pig farming by analyzing 105 fecal, environmental and slurry metagenomes from 38 farms. Results: Our results evidence a significantly higher abundance of antimicrobial resistance genes (ARGs) on intensive farms and a link between AMU and AMR to certain antimicrobial classes. We observed differences in the resistome across sample types, with a higher richness and dispersion of ARGs within environmental samples than on those from feces or slurry. Indeed, a deeper analysis revealed that differences among the three sample types were defined by taxa-ARGs associations. Interestingly, mobilome analyses revealed that the observed AMR differences between intensive and extensive farms could be linked to differences in the abundance of mobile genetic elements (MGEs). Thus, while there were no differences in the abundance of chromosomal-associated ARGs between intensive and extensive herds, a significantly higher abundance of integrons in the environment and plasmids, regardless of the sample type, was detected on intensive farms. Conclusions: Overall, this study shows how AMU, production system, and sample type influence, mainly through MGEs, the profile and dispersion of ARGs in pig production.S