80 research outputs found

    Interoperabilidad Gubernamental y Procedimientos Administrativos en una Unidad Desconcentrada de la Gerencia de Articulación Territorial

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    El avance permanente de la tecnología digital hace que las personas logren acceder a los datos necesarios y en el menor tiempo posible. Se consideró como objetivo general determinar la relación entre la interoperabilidad gubernamental y los procedimientos administrativos de una Unidad Desconcentrada de la Gerencia de Articulación Territorial, 2022. Se utilizó una metodología cuantitativa, básica, no experimental y descriptivo correlacional. Además, se utilizó la encuesta y el cuestionario que se aplicó a 62 trabajadores de la entidad estudiada. Los resultados revelaron que cuando la interoperabilidad gubernamental es inadecuada, los procedimientos administrativos son un 100% inadecuados. Asimismo, la interoperabilidad gubernamental y sus dimensiones mantienen una correlación positiva desde baja a muy fuerte, dado que obtuvieron un p=0.000<0.05, aceptando la hipótesis alterna. Se concluye que ambas variables están interconectadas entre sí, esto quiere decir que si una falla, la otra se verá afectada, es por ello que es sumamente necesario que la entidad debe contar con un sistema que le permita estar interconectado de manera directa con otras entidades del sector público ya que esto le admite atender de manera oportuna las solicitudes y reclamos de los usuarios, puesto que con ello aumentaría la eficiencia y eficacia en los procedimientos administrativos

    Metabolic engineering of Clostridium autoethanogenum for selective alcohol production

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    Gas fermentation using acetogenic bacteria such as Clostridium autoethanogenum offers an attractive route for production of fuel ethanol from industrial waste gases. Acetate reduction to acetaldehyde and further to ethanol via an aldehyde: ferredoxin oxidoreductase (AOR) and alcohol dehydrogenase has been postulated alongside the classic pathway of ethanol formation via a bi-functional aldehyde/alcohol dehydrogenase (AdhE). Here we demonstrate that AOR is critical to ethanol formation in acetogens and inactivation of AdhE led to consistently enhanced autotrophic ethanol production (up to 180%). Using ClosTron and allelic exchange mutagenesis, which was demonstrated for the first time in an acetogen, we generated single mutants as well as double mutants for both aor and adhE isoforms to confirm the role of each gene. The aor1+2 double knockout strain lost the ability to convert exogenous acetate, propionate and butyrate into the corresponding alcohols, further highlighting the role of these enzymes in catalyzing the thermodynamically unfavourable reduction of carboxylic acids into alcohols

    Regulatory targets of quorum sensing in Vibrio cholerae: evidence for two distinct HapR-binding motifs

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    The quorum-sensing pathway in Vibrio cholerae controls the expression of the master regulator HapR, which in turn regulates several important processes such as virulence factor production and biofilm formation. While HapR is known to control several important phenotypes, there are only a few target genes known to be transcriptionally regulated by HapR. In this work, we combine bioinformatic analysis with experimental validation to discover a set of novel direct targets of HapR. Our results provide evidence for two distinct binding motifs for HapR-regulated genes in V. cholerae. The first binding motif is similar to the motifs recently discovered for orthologs of HapR in V. harveyi and V. vulnificus. However, our results demonstrate that this binding motif can be of variable length in V. cholerae. The second binding motif shares common elements with the first motif, but is of fixed length and lacks dyad symmetry at the ends. The contributions of different bases to HapR binding for this second motif were demonstrated using systematic mutagenesis experiments. The current analysis presents an approach for systematically expanding our knowledge of the quorum-sensing regulon in V. cholerae and other related bacteria

    NO Dioxygenase Activity in Hemoglobins Is Ubiquitous In Vitro, but Limited by Reduction In Vivo

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    Genomics has produced hundreds of new hemoglobin sequences with examples in nearly every living organism. Structural and biochemical characterizations of many recombinant proteins reveal reactions, like oxygen binding and NO dioxygenation, that appear general to the hemoglobin superfamily regardless of whether they are related to physiological function. Despite considerable attention to “hexacoordinate” hemoglobins, which are found in nearly every plant and animal, no clear physiological role(s) has been assigned to them in any species. One popular and relevant hypothesis for their function is protection against NO. Here we have tested a comprehensive representation of hexacoordinate hemoglobins from plants (rice hemoglobin), animals (neuroglobin and cytoglobin), and bacteria (Synechocystis hemoglobin) for their abilities to scavenge NO compared to myoglobin. Our experiments include in vitro comparisons of NO dioxygenation, ferric NO binding, NO-induced reduction, NO scavenging with an artificial reduction system, and the ability to substitute for a known NO scavenger (flavohemoglobin) in E. coli. We conclude that none of these tests reveal any distinguishing predisposition toward a role in NO scavenging for the hxHbs, but that any hemoglobin could likely serve this role in the presence of a mechanism for heme iron re-reduction. Hence, future research to test the role of Hbs in NO scavenging would benefit more from the identification of cognate reductases than from in vitro analysis of NO and O2 binding

    The cytochrome bd-I respiratory oxidase augments survival of multidrug-resistant Escherichia coli during infection

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    Nitric oxide (NO) is a toxic free radical produced by neutrophils and macrophages in response to infection. Uropathogenic Escherichia coli (UPEC) induces a variety of defence mechanisms in response to NO, including direct NO detoxification (Hmp, NorVW, NrfA), iron-sulphur cluster repair (YtfE), and the expression of the NO-tolerant cytochrome bd-I respiratory oxidase (CydAB). The current study quantifies the relative contribution of these systems to UPEC growth and survival during infection. Loss of the flavohemoglobin Hmp and cytochrome bd-I elicit the greatest sensitivity to NO-mediated growth inhibition, whereas all but the periplasmic nitrite reductase NrfA provide protection against neutrophil killing and promote survival within activated macrophages. Intriguingly, the cytochrome bd-I respiratory oxidase was the only system that augmented UPEC survival in a mouse model after 2 days, suggesting that maintaining aerobic respiration under conditions of nitrosative stress is a key factor for host colonisation. These findings suggest that while UPEC have acquired a host of specialized mechanisms to evade nitrosative stresses, the cytochrome bd-I respiratory oxidase is the main contributor to NO tolerance and host colonisation under microaerobic conditions. This respiratory complex is therefore of major importance for the accumulation of high bacterial loads during infection of the urinary tract

    E. coli metabolic protein aldehydealcohol dehydrogenase-E binds to the ribosome: a unique moonlighting action revealed

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    It is becoming increasingly evident that a high degree of regulation is involved in the protein synthesis machinery entailing more interacting regulatory factors. A multitude of proteins have been identified recently which show regulatory function upon binding to the ribosome. Here, we identify tight association of a metabolic protein aldehyde-alcohol dehydrogenase E (AdhE) with the E. coli 70S ribosome isolated from cell extract under low salt wash conditions. Cryo-EM reconstruction of the ribosome sample allows us to localize its position on the head of the small subunit, near the mRNA entrance. Our study demonstrates substantial RNA unwinding activity of AdhE which can account for the ability of ribosome to translate through downstream of at least certain mRNA helices. Thus far, in E. coli, no ribosome-associated factor has been identified that shows downstream mRNA helicase activity. Additionally, the cryo-EM map reveals interaction of another extracellular protein, outer membrane protein C (OmpC), with the ribosome at the peripheral solvent side of the 50S subunit. Our result also provides important insight into plausible functional role of OmpC upon ribosome binding. Visualization of the ribosome purified directly from the cell lysate unveils for the first time interactions of additional regulatory proteins with the ribosom

    Discovering cis-Regulatory RNAs in Shewanella Genomes by Support Vector Machines

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    An increasing number of cis-regulatory RNA elements have been found to regulate gene expression post-transcriptionally in various biological processes in bacterial systems. Effective computational tools for large-scale identification of novel regulatory RNAs are strongly desired to facilitate our exploration of gene regulation mechanisms and regulatory networks. We present a new computational program named RSSVM (RNA Sampler+Support Vector Machine), which employs Support Vector Machines (SVMs) for efficient identification of functional RNA motifs from random RNA secondary structures. RSSVM uses a set of distinctive features to represent the common RNA secondary structure and structural alignment predicted by RNA Sampler, a tool for accurate common RNA secondary structure prediction, and is trained with functional RNAs from a variety of bacterial RNA motif/gene families covering a wide range of sequence identities. When tested on a large number of known and random RNA motifs, RSSVM shows a significantly higher sensitivity than other leading RNA identification programs while maintaining the same false positive rate. RSSVM performs particularly well on sets with low sequence identities. The combination of RNA Sampler and RSSVM provides a new, fast, and efficient pipeline for large-scale discovery of regulatory RNA motifs. We applied RSSVM to multiple Shewanella genomes and identified putative regulatory RNA motifs in the 5′ untranslated regions (UTRs) in S. oneidensis, an important bacterial organism with extraordinary respiratory and metal reducing abilities and great potential for bioremediation and alternative energy generation. From 1002 sets of 5′-UTRs of orthologous operons, we identified 166 putative regulatory RNA motifs, including 17 of the 19 known RNA motifs from Rfam, an additional 21 RNA motifs that are supported by literature evidence, 72 RNA motifs overlapping predicted transcription terminators or attenuators, and other candidate regulatory RNA motifs. Our study provides a list of promising novel regulatory RNA motifs potentially involved in post-transcriptional gene regulation. Combined with the previous cis-regulatory DNA motif study in S. oneidensis, this genome-wide discovery of cis-regulatory RNA motifs may offer more comprehensive views of gene regulation at a different level in this organism. The RSSVM software, predictions, and analysis results on Shewanella genomes are available at http://ural.wustl.edu/resources.html#RSSVM

    Regulation of expression of the Escherichia coli flavohaemoglobin and relationships with other globins

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    SIGLEAvailable from British Library Document Supply Centre-DSC:DXN018682 / BLDSC - British Library Document Supply CentreGBUnited Kingdo

    Experiencia de las enfermeras que trabajaron en el área de hospitalización COVID-19 durante la emergencia sanitaria en Lambayeque, 2021

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    Este estudio tuvo como objetivo: describir, analizar y comprender las experiencias de las enfermeras que trabajaron en el área de hospitalización COVID-19 durante la emergencia sanitaria. La investigación fue cualitativa con metodología descriptiva exploratoria. El tamaño de la muestra fue de 15 enfermeras que trabajaron en el área de hospitalización COVID-19, determinado por la técnica de la saturación y redundancia de datos. Se recolectó la información mediante entrevistas semiestructuradas a través de medios digitales, previo consentimiento informado, que fueron procesadas con el análisis de contenido. Se obtuvo como resultados 4 categorías: a) Miedo al contagio, impotencia ante la muerte y espiritualidad en el cuidado al paciente con COVID-19, b) Uso, adaptación y dificultades al implementar los protocolos preventivos durante la jornada laboral, c) Autocuidado y medidas de prevención en el hogar después de la jornada laboral para proteger a su familia de la COVID-19, y d) Aprendizajes entorno a la pandemia: cuidado técnico-humano y medidas preventivas. Se concluye, que las enfermeras entrevistadas tuvieron miedo a contagiarse y contagiar a su familia, sentían impotencia por ver morir a sus pacientes, pero incrementaron su fe en Dios. Se adaptaron a los protocolos para usar el equipo de protección personal, a pesar que tuvieron dificultades y algunas se contagiaron durante su retiro. Mejoraron su alimentación, la actividad física, y aplicaron estrictamente las medidas preventivas ante la COVID-19 en sus hogares. Aprendieron a ser más empáticos con los pacientes y sus compañeros de trabajo
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