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Introducción: Los ictus se producen ante una interrupción del flujo cerebral normal. Esto puede ocasionar graves secuelas en la vida de los pacientes que lo sufren. Es fundamental un rápido reconocimiento de los signos y síntomas y una rápida alerta a los servicios de emergencias para su traslado al centro hospitalario de referencia, lo que reducirá las posteriores secuelas en la salud del paciente. Para proporcionar una respuesta rápida y eficaz, existen protocolos donde se establecen directrices a la hora de tratar a estos pacientes. Objetivos: Proporcionar un conjunto de recomendaciones y un protocolo actualizado, basado en distintos protocolos utilizados en diferentes Comunidades Autónomas (CCAA), para el manejo del paciente ictus en fase aguda durante la asistencia extrahospitalaria e intrahospitalaria y proporcionar información acerca de la patología, cómo reconocerla y concienciar a los lectores acerca del importante impacto que provocan en la sociedad. Métodos: Se realizaron varias estrategias de búsqueda. La primera fue acudir a las bases de datos Google Académico, Medline, Elsevier; además de las páginas oficiales del Ministerio de Sanidad, el Instituto Nacional de Estadística (INE) y el protocolo de la Sociedad Española de Medicina de Urgencias y Emergencias (SEMES) en los pacientes con ictus. La segunda consistió en la búsqueda de protocolos de los Códigos Ictus (CI) implantados en diferentes CCAA. Resultados: Se realizaron tablas comparativas entre las diferentes CCAA de los apartados siguientes: Criterios de activación del CI; Primera asistencia extrahospitalaria; Tratamientos realizados durante el traslado; Asistencia hospitalaria y Cuidados realizados tras el ingreso del paciente en Urgencias. Conclusiones: Se establecen puntos de mejora no presentes en el CI gallego como; la utilización de escalas funcionales, la potenciación del papel de enfermería durante todo el proceso asistencial mediante la creación de un plan de cuidados específicos para el u, Introdución: Os ictus prodúcense ante unha interrupción do fluxo cerebral normal. Isto pode ocasionar graves secuelas na vida dos pacientes que o sofren. É fundamental un rápido recoñecemento dos signos e síntomas e unha rápida alerta aos servizos de emerxencias para o seu traslado ao centro hospitalario de referencia, o que reducirá as posteriores secuelas na saúde do paciente. Para proporcionar unha resposta rápida e eficaz, existen protocolos onde se establecen directrices á hora de tratar a estes pacientes. Obxectivos: Proporcionar un conxunto de recomendacións e un protocolo actualizado, completo e baseado en distintos protocolos utilizados en diferentes Comunidades Autónomas (CCAA), para o manexo do paciente ictus en fase aguda durante a asistencia extrahospitalaria e intrahospitalaria e proporcionar información acerca de qué é o ictus, como recoñecelo e concienciar aos lectores achega do importante impacto que provocan na sociedade. Métodos: Realizáronse varias estratexias de procura. A primeira foi acodir as bases de datos Google Académico, Medline, Elsevier; ademais de las páxinas oficiais como o Ministerio de Sanidade, o Instituto Nacional de Estatística (INE) e o protocolo do SEMES nos pacientes con ictus. A segunda consistiu na procura de protocolos dos Códigos Ictus (CI) implantados en diferentes CCAA. Resultados: Realizáronse táboas comparativas entre as diferentes CCAA dos apartados seguintes: Criterios de activación do CI; Primeira asistencia extrahospitalaria; Tratamentos realizados durante o traslado; Asistencia hospitalaria e Coidados realizados tras o ingreso do paciente en Urxencias. Conclusións: Establécense puntos de mellora non presentes no CI galego como; a utilización de escalas funcionais, a potenciación do papel de enfermería durante todo o proceso asistencial mediante a creación dun plan de coidados específicos para o uso destes profesionais ou a elaboración dun protocolo ictus pediátrico, entre outras, Introduction: Strokes are caused by an interruption of normal brain flow. This can cause serious sequelae in the life of the patients who suffer it. A quick recognition of the signs and symptoms and a rapid alert to the emergency services for transfer to the hospital of reference is essential, which will reduce the subsequent sequelae in the patient's health. In order to provide a rapid and effective response, there are protocols that establish guidelines for the treatment of these patients. Objectives: To provide a set of recommendations and an updated, complete protocol based on different protocols used in different Autonomous Communities, for the management of stroke patients in the acute phase during out-of-hospital and in-hospital care and to provide information about what stroke is, how to recognize it, and to raise awareness of the important impact it has on society. Methods: Several search strategies were used. The first was in data bases such as Google Scholar,Medline, Elsevier; in addition to official pages such as the Ministerio de Sanidad, the Instituto Nacional de Estadística (INE) and SEME´s protocol w patients . The second consisted of a search for stroke code protocols implemented in different Autonomous Communities. Results: Comparative tables were drawn up for the following sections between the different Autonomous Communities: Criteria for activation of the Stroke Code; First out-of-hospital care;Treatments performed during transfer; In-hospital care and Care performed after admission of the patient to the Emergency Department. Conclusions: Points for improvement not present in the Galician SC are established, such as the use of functional scales, the strengthening of the role of nurses throughout the care process by creating a specific care plan for the use of these professionals or the development of a pediatric stroke protocol, among others

El incremento de la demanda de alimentos y el cambio climático están obligando a buscar alternativas sostenibles que mejoren la productividad agrícola. Además de los rizobios, existen otras bacterias (Plant Growth Promoting Bacteria; PGPB) menos estudiadas que mejoran el acceso de las plantas a los nutrientes o su tolerancia al estrés incluyendo especies de alto interés comercial como los cereales. Este trabajo se centra en el estudio de la comunidad cultivable y no-cultivable de bacterias endófitas aisladas de diferentes variedades de trigo relacionadas con su domesticación obtenidas en distintas localizaciones geográficas. Nuestros análisis metataxonómicos muestran que la estructura de la comunidad de distintas semillas se encuentra conservada y dominada por un único género bacteriano perteneciente al grupo de las Enterobacterias. Este género forma parte del microbioma core del trigo, ya que es el único presente en todas sus variedades y tejidos, independientemente de su localización geográfica. Además, su abundancia aumenta proporcionalmente de suelo a raíz, tallo y espiga, acumulándose finalmente en las semillas. Este enriquecimiento sugiere un papel en la transferencia vertical de estas bacterias endófitas a través de las semillas. Para comprobar esta hipótesis, flores y semillas de cereales (trigo y Lolium) y Arabidopsis fueron inoculadas con aislados de este género marcados con un plásmido que codifica la enzima ß-glucoronidasa. Semillas de la siguiente progenie fueron germinadas para comprobar, mediante revelado con el sustrato cromogénico X-gluc, la presencia del endófito únicamente en cereales, confirmando su transmisión vertical a través de la línea germinal de forma específica.

Bacteria: are powerful models for understanding how cells divide : and accomplish global regulatory programs. In Caulobacter crescentus, a cascade of essential master regulators supervises the correct and sequential activation of DNA replication, cell division, and development of different cell types. Among them, the response regulator CtrA plays a crucial role coordinating all those functions. Here, for the first time, we describe the role of a novel factor named CcnA (cell cycle noncoding RNA A), a cell cycle–regulated noncoding RNA (ncRNA) located at the origin of replication, presumably activated by CtrA, and responsible for the accumulation of CtrA itself. In addition, CcnA may be also involved in the inhibition of translation of the S-phase regulator, GcrA, by interacting with its 5 untranslated region (5 UTR). Performing in vitro experiments and mutagenesis, we propose a mechanism of action of CcnA based on liberation (ctrA) or sequestration (gcrA) of their ribosome-binding site (RBS). Finally, its role may be conserved in other alphaproteobacterial species, such as Sinorhizobium meliloti, representing indeed a potentially conserved process modulating cell cycle in Caulobacterales and Rhizobiales.

Genetic interventions on microbiomes, for clinical or biotechnological purposes, remain challenging. Conjugation-based delivery of genetic cargo is still unspecific and limited by low conjugation rates. Here we report an approach to overcome these problems, based on a synthetic bacterial adhesion system. Mating assemblers consist on a synthetic adhesion formed by the expression on the surface of donor and target cells of specific nanobodies (Nb) and their cognate antigen (Ag). The Nb–Ag bridge increased 1–3 logs transfer of a variety of plasmids, especially in liquid media, confirming that cell-cell docking is a main determinant limiting mating efficiency. Synthetic cell-to-cell adhesion allows efficient conjugation to targeted recipients, enhancing delivery of desired genes to a predefined subset of prey species, or even specific pathogenic strains such as enterohemorrhagic Escherichia coli (EHEC), within a bacterial community. The synthetic conjugation enhancer presented here optimizes plasmid delivery by selecting the target hosts with high selectivity.

The rhizosphere and rhizoplane are nutrient-rich but selective environments for the root microbiome. Here, we deciphered a posttranscriptional network regulated by the homologous trans-small RNAs (sRNAs) AbcR1 and AbcR2, which rewire the metabolism of the nitrogen-fixing a-rhizobium Sinorhizobium meliloti during preinfection stages of symbiosis with its legume host alfalfa. The LysR-type regulator LsrB, which transduces the cell redox state, is indispensable for AbcR1 expression in actively dividing bacteria, whereas the stress-induced transcription of AbcR2 depends on the alternative s factor RpoH1. MS2 affinity purification coupled with RNA sequencing unveiled exceptionally large and overlapping AbcR1/2 mRNA interactomes, jointly representing;6% of the S. meliloti protein-coding genes. Most mRNAs encode transport/metabolic proteins whose translation is silenced by base pairing to two distinct anti-Shine Dalgarno motifs that function independently in both sRNAs. A metabolic model-aided analysis of the targetomes predicted changes in AbcR1/2 expression driven by shifts in carbon/nitrogen sources, which were confirmed experimentally. Low AbcR1/2 levels in some defined media anticipated overexpression growth phenotypes linked to the silencing of specific mRNAs. As a proof of principle, we confirmed AbcR1/2-mediated downregulation of the L-amino acid AapQ permease. AbcR1/2 interactomes are well represented in rhizosphere-related S. meliloti transcriptomic signatures. Remarkably, a lack of AbcR1 specifically compromised the ability of S. meliloti to colonize the root rhizoplane. The AbcR1 regulon likely ranks the utilization of available substrates to optimize metabolism, thus conferring on S. meliloti an advantage for efficient rhizosphere/rhizoplane colonization. AbcR1 regulation is predicted to be conserved in related a-rhizobia, which opens unprecedented possibilities for engineering highly competitive biofertilizers. IMPORTANCE Nitrogen-fixing root nodule symbioses

Lyme carditis is an extracutaneous manifestation of Lyme disease characterized by episodes of atrioventricular block of varying degrees and additional, less reported cardiomyopathies. The molecular changes associated with the response to Borrelia burgdorferi over the course of infection are poorly understood. Here, we identify broad transcriptomic and proteomic changes in the heart during infection that reveal a profound down-regulation of mitochondrial components. We also describe the long-term functional modulation of macrophages exposed to live bacteria, characterized by an augmented glycolytic output, increased spirochetal binding and internalization, and reduced inflammatory responses. In vitro, glycolysis inhibition reduces the production of tumor necrosis factor (TNF) by memory macrophages, whereas in vivo, it produces the reversion of the memory phenotype, the recovery of tissue mitochondrial components, and decreased inflammation and spirochetal burdens. These results show that B. burgdorferi induces long-term, memory-like responses in macrophages with tissue-wide consequences that are amenable to be manipulated in vivo.

Small non-coding RNAs (sRNAs) are ubiquitous components of bacterial adaptive regulatory networks underlying stress responses and chronic intracellular infection of eukaryotic hosts. Thus, sRNA-mediated regulation of gene expression is expected to play a major role in the establishment of mutualistic root nodule endosymbiosis between nitrogen-fixing rhizobia and legume plants. However, knowledge about this level of genetic regulation in this group of plant-interacting bacteria is still rather scarce. Here, we review insights into the rhizobial non-coding transcriptome and sRNA-mediated post-transcriptional regulation of symbiotic relevant traits such as nutrient uptake, cell cycle, quorum sensing, or nodule development. We provide details about the transcriptional control and protein-assisted activity mechanisms of the functionally characterized sRNAs involved in these processes. Finally, we discuss the forthcoming research on riboregulation in legume symbionts.

Small non-coding RNAs (sRNAs) are ubiquitous components of regulatory netwoks underlying virtually any adaptive response in bacteria. As regulators of gene expression, sRNAs accumulate differentially in the cell in response to external cues and their main activity mechanism involves protein-assisted base-pairing interactions with short and discontinuous stretches of complementary nucleotides usually mapping to the translation initiation region of multiple trans-encoded target mRNAs. In most cases, this antisense interaction promotes post-transcriptional gene silencing by blocking of translation and degradation of the target mRNAs by cellular RNases (reviewed in Jiménez-Zurdo et al. 2013). In our laboratory, we are interested in the characterization of the non-coding transcriptome of the alfalfa symbiont Sinorhizobium meliloti. Deletion of one of the hundreds of sRNA genes identified in this bacterium alters nodule organogenesis and compromises nitrogen-fixation efficiency on alfalfa roots (Robledo et al, 2017). According to this symbiotic phenotype, this sRNA was named NfeR1 (Nodule Formation Efficiency RNA). Computational predictions, some of which have been verified experimentally, suggested that NfeR1 uses redundantly three identical anti-Shine Dalgarno (aSD) motifs for targeting and down-regulation of multiple mRNAs coding for the periplasmic component of ABC transport systems. Northern blot probing and transcription tracking with promoter-eGFP fusions uncovered maximum NfeR1 accumulation upon an osmotic upshift in free-living bacteria and during all steps of the symbiotic interaction. The strength and differential regulation of nfeR1 transcription is conferred by a motif that is conserved in the promoter regions of its ¿-proteobacterial homologs. Pull-down assays revealed that the NfeR1 promoter might bind the global nitrogen regulator NtrC and the LysR type transcriptional regulator LsrB (LysR Symbiotic Regulator). In the communication, we will provide novel i

Primary infection of legumes by rhizobia involves the controlled localized enzymatic breakdown of cell walls at root hair tips. Previous studies determined the role of rhizobial CelC2 cellulase in different steps of the symbiotic interaction Rhizobium leguminosarum-Trifolium repens. Recent findings also showed that CelC2 influences early signalling events in the Ensifer meliloti-Medicago truncatula interaction. Here, we have monitored the root hair phenotypes of two legume plants, T. repens and M. sativa, upon inoculation with strains of their cognate and non-cognate rhizobial species, R. leguminosarum bv trifolii and E. meliloti, (over)expressing the CelC2 coding gene, celC. Regardless of the host, CelC2 specifically elicited ‘hole-on-the-tip’ events (Hot phenotype) in the root hair apex, consistent with the role of this endoglucanase in eroding the noncrystalline cellulose found in polarly growing cell walls. Overproduction of CelC2 also increased root hair tip redirections (RaT phenotype) events in both cognate and non-cognate hosts. Interestingly, heterologous celC expression also induced non-canonical alterations in ROS (Reactive Oxygen Species) homeostasis at root hair tips of Trifolium and Medicago. These results suggest the concurrence of shared unspecific and host-related plant responses to CelC2 during early steps of symbiotic rhizobial infection. Our data thus identify CelC2 cellulase as an important determinant of events underlying early infection of the legume host by rhizobia.

Adjustment of cell cycle progression is crucial for bacterial survival and adaptation under adverse conditions. However, the understanding of modulation of cell cycle control in response to environmental changes is rather incomplete. In a-proteobacteria, the broadly conserved cell cycle master regulator CtrA underlies multiple levels of control, including coupling of cell cycle and cell differentiation. CtrA levels are known to be tightly controlled through diverse transcriptional and post-translational mechanisms. Here, small RNA (sRNA)-mediated post-transcriptional regulation is uncovered as an additional level of CtrA fine-tuning. Computational predictions as well as transcriptome and proteome studies consistently suggested targeting of ctrA and the putative cold shock chaperone cspA5 mRNAs by the trans-encoded sRNA (trans-sRNA) GspR (formerly SmelC775) in several Sinorhizobium species. GspR strongly accumulated in the stationary growth phase, especially in minimal medium (MM) cultures. Lack of the gspR locus confers a fitness disadvantage in competition with the wild type, while its overproduction hampers cell growth, suggesting that this riboregulator interferes with cell cycle progression. An eGFP-based reporter in vivo assay, involving wild-type and mutant sRNA and mRNA pairs, experimentally confirmed GspR-dependent post-transcriptional down-regulation of ctrA and cspA5 expression, which most likely occurs through base-pairing to the respective mRNA. The energetically favored secondary structure of GspR is predicted to comprise three stem-loop domains, with stem-loop 1 and stem-loop 3 targeting ctrA and cspA5 mRNA, respectively. Moreover, this work reports evidence for post-transcriptional control of ctrA by CspA5. Thus, this regulation and GspR-mediated post-transcriptional repression of ctrA and cspA5 expression constitute a coherent feed-forward loop, which may enhance the negative effect of GspR on CtrA levels. This novel regulatory circuit involving the

Members of the ribonuclease (RNase) III family of enzymes are metal-dependent double-strand specific endoribonucleases. They are ubiquitously found and eukaryotic RNase III-like enzymes include Dicer and Drosha, involved in RNA processing and RNA interference. In this work, we have addressed the primary characterization of RNase III from the symbiotic nitrogen-fixing a-proteobacterium Sinorhizobium meliloti. The S. meliloti rnc gene does encode an RNase III-like protein (SmRNase III), with recognizable catalytic and double-stranded RNA (dsRNA)-binding domains that clusters in a branch with its a-proteobacterial counterparts. Purified SmRNase III dimerizes, is active at neutral to alkaline pH and behaves as a strict metal cofactor-dependent double-strand endoribonuclease, with catalytic features distinguishable from those of the prototypical member of the family, the Escherichia coli ortholog (EcRNase III). SmRNase III prefers Mn2+ rather than Mg as metal cofactor, cleaves the generic structured R1.1 substrate at a site atypical for RNase III cleavage, and requires higher cofactor concentrations and longer dsRNA substrates than EcRNase III for optimal activity. Furthermore, the ultraconserved E125 amino acid was shown to play a major role in the metal-dependent catalysis of SmRNase III. SmRNase III degrades endogenous RNA substrates of diverse biogenesis with different efficiency, and is involved in the maturation of the 23S rRNA. SmRNase III loss-of-function neither compromises viability nor alters morphology of S. meliloti cells, but influences growth, nodulation kinetics, the onset of nitrogen fixation and the overall symbiotic efficiency of this bacterium on the roots of its legume host, alfalfa, which ultimately affects plant growth. Our results support an impact of SmRNase III on nodulation and symbiotic nitrogen fixation in plants.

The infection of legume plants by rhizobia is tightly regulated to ensure accurate bacterial penetration, infection, and development of functionally efficient nitrogen-fixing root nodules. Rhizobial Nod factors (NF) have key roles in the elicitation of nodulation signaling. Infection of white clover roots also involves the tightly regulated specific breakdown of the noncrystalline apex of cell walls in growing root hairs, which is mediated by Rhizobium leguminosarum bv. trifolii cellulase CelC2. Here, we have analyzed the impact of this endoglucanase on symbiotic signaling in the model legume Medicago truncatula. Ensifer meliloti constitutively expressing celC gene exhibited delayed nodulation and elicited aberrant ineffective nodules, hampering plant growth in the absence of nitrogen. Cotreatment of roots with NF and CelC2 altered Ca spiking in root hairs and induction of the early nodulin gene ENOD11. Our data suggest that CelC2 alters early signaling between partners in the rhizobia-legume interaction.