Insights into a Novel blaKPC-2-Encoding IncP-6 Plasmid Reveal Carbapenem-Resistance Circulation in Several Enterobacteriaceae Species from Wastewater and a Hospital Source in Spain

Untreated wastewater, particularly from hospitals and other healthcare facilities, is considered to be a reservoir for multidrug-resistant bacteria. However, its role in the spread of antibiotic resistances in the human population remains poorly investigated. We used whole genome sequencing (WGS) to analyze 25 KPC-2-producing Enterobacteriaceae isolates from sewage water collected during a 3-year period and three clinical Citrobacter freundii isolates from a tertiary hospital in the same collection area in Spain. We detected a common, recently described, IncP-6 plasmid carrying the gene blaKPC-2 in 21 isolates from both sources. The plasmid was present in diverse environmental bacterial species of opportunistic pathogens such as C. freundii, Enterobacter cloacae, Klebsiella oxytoca, and Raoultella ornithinolytica. The 40,186 bp IncP-6 plasmid encoded 52 coding sequences (CDS) and was composed of three uniquely combined regions that were derived from other plasmids recently reported in different countries of South America. The region harboring the carbapenem resistance gene (14 kb) contained a Tn3 transposon disrupted by an ISApu-flanked element and the core sequence composed by ISKpn6 / blaKPC-2 / ?blaTEM-1 / ISKpn27. We document here the presence of a novel promiscuous blaKPC-2 plasmid circulating in environmental bacteria in wastewater and human populations

Staphylococcus aureus carrying mecC gene in animals and urban wastewater, Spain

Letter to the edito

Uncovering a Novel Functional Interaction Between Adult Hepatic Progenitor Cells, Inflammation and EGFR Signaling During Bile Acids-Induced Injury

Chronic cholestatic damage is associated to both accumulation of cytotoxic levels of bile acids and expansion of adult hepatic progenitor cells (HPC) as part of the ductular reaction contributing to the regenerative response. Here, we report a bile acid-specific cytotoxic response in mouse HPC, which is partially impaired by EGF signaling. Additionally, we show that EGF synergizes with bile acids to trigger inflammatory signaling and NLRP3 inflammasome activation in HPC. Aiming at understanding the impact of this HPC specific response on the liver microenvironment we run a proteomic analysis of HPC secretome. Data show an enrichment in immune and TGF-beta regulators, ECM components and remodeling proteins in HPC secretome. Consistently, HPC-derived conditioned medium promotes hepatic stellate cell (HSC) activation and macrophage M1-like polarization. Strikingly, EGF and bile acids co-treatment leads to profound changes in the secretome composition, illustrated by an abolishment of HSC activating effect and by promoting macrophage M2-like polarization. Collectively, we provide new specific mechanisms behind HPC regulatory action during cholestatic liver injury, with an active role in cellular interactome and inflammatory response regulation. Moreover, findings prove a key contribution for EGFR signaling jointly with bile acids in HPC-mediated actions

Lack of EGFR catalytic activity in hepatocytes improves liver regeneration following DDC-induced cholestatic injury by promoting a pro-restorative inflammatory response

Despite the well-known hepatoprotective role of the epidermal growth factor receptor (EGFR) pathway upon acute damage, its specific actions during chronic liver disease, particularly cholestatic injury, remain ambiguous and unresolved. Here, we analyzed the consequences of inactivating EGFR signaling in the liver on the regenerative response following cholestatic injury. For that, transgenic mice overexpressing a dominant negative mutant human EGFR lacking tyrosine kinase activity (ΔEGFR) in albumin-positive cells were submitted to liver damage induced by 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC), an experimental model resembling human primary sclerosing cholangitis. Our results show an early activation of EGFR after 1–2 days of a DDC-supplemented diet, followed by a signaling switch-off. Furthermore, ΔEGFR mice showed less liver damage and a more efficient regeneration following DDC injury. Analysis of the mechanisms driving this effect revealed an enhanced activation of mitogenic/survival signals, AKT and ERK1/2-MAPKs, and changes in cell turnover consistent with a quicker resolution of damage in response to DDC. These changes were concomitant with profound differences in the profile of intrahepatic immune cells, consisting of a shift in the M1/M2 balance towards M2 polarity, and the Cd4/Cd8 ratio in favor of Cd4 lymphocytes, overall supporting an immune cell switch into a pro-restorative phenotype. Interestingly, ΔEGFR livers also displayed an amplified ductular reaction, with increased expression of EPCAM and an increased number of CK19-positive ductular structures in portal areas, demonstrating an overexpansion of ductular progenitor cells. In summary, our work supports the notion that hepatocyte-specific EGFR activity acts as a key player in the crosstalk between parenchymal and non-parenchymal hepatic cells, promoting the pro-inflammatory response activated during cholestatic injury and therefore contributing to the pathogenesis of cholestatic liver disease. © 2022 The Pathological Society of Great Britain and Ireland.This work was supported by the Ministry of Science,Innovation and Universities (MICIU) and AgenciaEstatal de Investigacion (AEI), Spain (co-funded byFEDER funds/Development Fund–a Way to BuildEurope): RTI2018-099098-B-100 to AS/BH andRTI2018-094052-B-100 to AMV; and the RamonAreces Foundation: 20th National Competition forScientific and Technical Research in Life and MatterScience (2020) to IF. NL and JGS were recipients ofresearch assistant contracts linked to grant SAF2015-69145-R and RTI2018-099098-B-100, respectively. CMR was the recipient of a researchcontract (PEJD-2019-POST/BMD-16090) from the Education, Universities, Research and Spokesperson Counseling of the Community of Madrid

EducaFarma 11.0

Memoria ID2022-036. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2022-2023

Educafarma 10.0

Memoria ID-030. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2021-2022

EducaFarma 9.0

Memoria ID-020 Ayudas de la Universidad de Salamanca para la innovación docente, curso 2020-2021

Manual de análisis para el control de calidad de la cerveza en los laboratorios del Departamento de Ingeniería Química y Ambiental

Este trabajo es la continuación del Trabajo Fin de Grado: Análisis fisicoquímicos para el control de calidad en la producción de cerveza (1), por lo cual, parte de la información teórica inicial se basa en ese proyecto. Se pretende realizar una extensión realizando la parte práctica en los laboratorios del Departamento de Ingeniería Química y Ambiental de la Escuela Técnica Superior de Ingeniería de Sevilla. El objetivo de este documento es hacer una investigación sobre qué parámetros, según el BOE, se pueden determinar y qué técnicas se pueden utilizar en dichos laboratorios, por qué se realizan y describir, tanto los materiales como el proceso, de forma exhaustiva para que este documento pueda servir como guía para el procedimiento de éstos. Para llegar a este objetivo, el documento consta inicialmente de una introducción a la cerveza, en la cual se resume la forma de elaboración y las distintas formas existentes de clasificar la cerveza. A continuación, se realiza un resumen de los parámetros importantes para la calidad de la cerveza, tanto parámetros obligatorios como aconsejables a determinar. Seguidamente, se expondrán los análisis realizables en las instalaciones y una explicación de las técnicas utilizadas y su procedimiento. Así mismo, los parámetros y técnicas que no puedan ser realizadas, serán explicadas y se justificarán los diversos motivos por los que no es posible y se explicará que es necesario para poder llevarlos a cabo. Como último análisis, se expone una solución a las técnicas no realizables, para ello se explica que equipos o materiales se deben adquirir y la inversión que supone.This work is the continuation of the End of Degree Work: Physical-chemical analysis for quality control in beer production (1), so, part of the initial theoretical information is based on that project. It is intended to carry out an extension by performing the practical part in the laboratories of the Department of Chemical and Environmental Engineering of the Escuela Técnica Superior de Ingeniería in Seville. The aim of this document is to make an investigation on which parameters, according to the BOE, can be determined and which techniques can be used in those laboratories, why they are made and to describe, both the materials and the process, in a comprehensive way so that this document can serve as a guide for their procedure. In order to achieve this objective, the document initially consists of an introduction to beer, in which the way it is brewed, and the different existing ways of classifying beer are summarized. Afterwards, a summary is made of the important parameters for the quality of the beer, both mandatory and advisable parameters to be determined. Afterwards, the analyses that can be carried out in the facilities will be presented, along with an explanation of the techniques used and their procedure. Likewise, the parameters and techniques that cannot be carried out will be explained and the different reasons why this is not possible will be justified and it will be explained what is necessary to carry them out. As a final analysis, a solution to the unfeasible techniques is presented, explaining what equipment or materials must be acquired and the investment involved.Universidad de Sevilla. Grado en Ingeniería Químic

Beta-lactamasas de espectro extendido en "Enterobacteriaceae": bases genéticas y epidemiológicas de su diseminación en diferentes compartimentos

Esta tesis doctoral tiene como misión profundizar en el conocimiento de las beta-lactamasas de espectro extendido (BLEE), de los elementos genéticos que contienen los genes correspondientes y de los microorganismos que las producen con un abordaje epidemiológico y de aplicación de diferentes técnicas de microbiología molecular. Esta aproximación es esencial para comprender la importancia epidemiológica y algunos de los factores que han contribuido a la diseminación de este mecanismo de resistencia. Su estudio a partir de colecciones de microorganismos obtenidos de muestras clínicas de pacientes, de estudios ecológicos de colonización (portadores) tanto en el hospital como en la comunidad y de muestras de aguas y alimentos responde a la necesidad de analizar este problema y generar conocimiento científico para ofrecer respuestas y establecer estrategias ante el problema de salud pública que constituyen los microorganismos productores de BLEE. Nuestra hipótesis de trabajo se fundamentó en establecer la importancia del compartimento intestinal de los pacientes hospitalizados e individuos de la comunidad como reservorio de microorganismos productores de BLEE y como indicador de la epidemiología y los tipos de enzimas encontrados en la clínica. El objetivo general es analizar la epidemiología, la estructura poblacional y profundizar en el estudio de los elementos genéticos implicados en la diseminación de enterobacterias productoras de BLEE en el ámbito hospitalario (Hospital Universitario Ramón y Cajal) y en el extrahospitalario (Área sanitaria 4 de la Comunidad de Madrid)