Shared apical sorting of anion exchanger isoforms AE2a, AE2b1, and AE2b2 in primary hepatocytes

AE2 (SLC4A2) is the member of the Na(+)-independent anion exchanger (AE) family putatively involved in the secretion of bicarbonate to bile. In humans, three variants of AE2 mRNA have been described: the full-length transcript AE2a (expressed from the upstream promoter in most tissues), and alternative transcripts AE2b(1) and AE2b(2) (driven from alternate promoter sequences in a tissue-restricted manner, mainly in liver and kidney). These transcripts would result in AE protein isoforms with short N-terminal differences. To ascertain their translation, functionality, and membrane sorting, we constructed expression vectors encoding each AE2 isoform fused to GFP at the C-terminus. Transfected HEK293 cells showed expression of functional GFP-tagged AE2 proteins, all three isoforms displaying comparable AE activities. Primary rat hepatocytes transfected with expression vectors and repolarized in a collagen-sandwich configuration showed a microtubule-dependent apical sorting of each AE2 isoform. This shared apical sorting is liver-cell specific, as sorting of AE2 isoforms was basolateral in control experiments on polarized kidney MDCK cells. Hepatocytic apical targeting of AE2 isoforms suggests that they all may participate in the canalicular secretion of bicarbonate to bile

G protein-coupled estrogen receptor activation by bisphenol-A disrupts the protection from apoptosis conferred by the estrogen receptors ERα and ERβ in pancreatic beta cells

17β-estradiol protects pancreatic β-cells from apoptosis via the estrogen receptors ERα, ERβ and GPER. Conversely, the endocrine disruptor bisphenol-A (BPA), which exerts multiple effects in this cell type via the same estrogen receptors, increased basal apoptosis. The molecular-initiated events that trigger these opposite actions have yet to be identified. We demonstrated that combined genetic downregulation and pharmacological blockade of each estrogen receptor increased apoptosis to a different extent. The increase in apoptosis induced by BPA was diminished by the pharmacological blockade or the genetic silencing of GPER, and it was partially reproduced by the GPER agonist G1. BPA and G1-induced apoptosis were abolished upon pharmacological inhibition, silencing of ERα and ERβ, or in dispersed islet cells from ERβ knockout (BERKO) mice. However, the ERα and ERβ agonists PPT and DPN, respectively, had no effect on beta cell viability. To exert their biological actions, ERα and ERβ form homodimers and heterodimers. Molecular dynamics simulations together with proximity ligand assays and coimmunoprecipitation experiments indicated that the interaction of BPA with ERα and ERβ as well as GPER activation by G1 decreased ERαβ heterodimers. We propose that ERαβ heterodimers play an antiapoptotic role in beta cells and that BPA- and G1-induced decreases in ERαβ heterodimers lead to beta cell apoptosis. Unveiling how different estrogenic chemicals affect the crosstalk among estrogen receptors should help to identify diabetogenic endocrine disruptors.This work was supported by Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación (AEI) and Fondo Europeo de Desarrollo Regional (FEDER) grants BPU2017-86579-R (AN), PID2020-117294RB-I00 (AN, JM-P), Generalitat Valenciana PROMETEO II/2020/006 (AN) and European Union’s Horizon 2020 research and innovation programme under grant agreement GOLIATH No. 825489 (AN). Author laboratories hold grants from Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación y Fondo Europeo de Desarrollo Regional (FEDER) RTI2018-096724-B-C21 (J-AE) and PID2020-117569RA-I00 (LM). PROMETEO/2016/006 (J-AE) and SEJI/2018/023 (LM) supported by Generalitat Valenciana, Spain. Robert A. Welch Foundation (grant E-0004) (J-AG). CIBERDEM is an initiative of the Instituto de Salud Carlos III

Risk factors for mortality caused by pseudomonas aeruginosa infection in hospitalized patients with oncologic diagnosis in three cities of Colombia

Pseudomonas aeruginosa es una bacteria oportunista Gram negativa particularmente eficiente en la adquisición de mecanismos de resistencia y de alta prevalencia en infecciones nosocomiales en pacientes oncológicos. Objetivo: identificar los factores de riesgo para mortalidad en pacientes oncológicos con aislamiento de P. aeruginosa. Metodología: estudio descriptivo, la población de estudio fueron los casos reportados con aislamiento de P. aeruginosa en el servicio de hospitalización de Oncólogos de Occidente en Pereira, Armenia y Manizales durante el año 2015. Se realizaron análisis univariados y multivariados; la supervivencia se estableció según el método de Kaplan-Meier. Se estableció un valor de p <0.05. Se usó el software STATA. Se tuvo aval de bioética de la Universidad Tecnológica de Pereira. Resultados: se estudió 41 casos confirmados de cultivos positivos de P. aeruginosa. El sexo masculino (46.3%), anemia (46.3%), neutropenia febril (41%), trombocitopenia (29.3%) y haber sido hospitalizado en la unidad de cuidados intensivos (29.3%) fueron asociados estadísticamente con mayor mortalidad (p=0.019); con estos resultados se diseñó una escala de riesgo (alfa de Cronbach =0.72). Los pacientes con cuatro de estas exposiciones mostraron mayor riesgo de mortalidad al egreso hospitalario con una sensibilidad del 68% y especificidad del 90%. La P. aeruginosa presentó resistencia a cefepime (36.6%) y a aztreonam (34.1%), mientras que la letalidad global fue del 26.8%. Conclusión: El sexo masculino, la coexistencia de anemia, trombocitopenia, y neutropenia febril, así como la estancia en la unidad de cuidados intensivos aumentan la mortalidad en los pacientes oncológicos infectados con P. aeruginosa.Pseudomonas aeruginosa is a Gram-negative and rod-shape opportunistic bacterium that is particularly efficient in the acquisition of resistance mechanisms and its high prevalence in nosocomial infections in cancer patients. Objective: To identify risk factors for mortality in cancer patients with P. aeruginosa infection. Methodology: A descriptive study was carried out in patients with P. aeruginosa infection during the hospitalization service of “Oncólogos de Occidente” in Pereira, Armenia and Manizales during 2015. Univariate and multivariate analyzes were performed. The survival analysis was established according to the Kaplan-Meier method. A value of p<0.05 was established for it. The analyses were examined with the STATA software. This study was endorsed by the bioethics committee of the “Universidad Tecnológica de Pereira”. Results: Fourty-one patients with positive culture for P. aeruginosa were studied. Males (46.3%), anemia (46.3%), febrile neutropenia (41%), thrombocytopenia (29.3%) and previous hospitalization in an intensive care unit (29.3%) were associated with higher mortality risk (p = 0.019); a risk scale was designed with these factors (Cronbach´s alpha = 0.72). Patients who presented four of these exposures were at higher risk of mortality with a sensitivity of 68% and specificity of 90% at the moment of discharge. P. aeruginosa showed 36.6% of resistance to cefepime, 34.1% to aztreonam, the mortality rate was 26.8%. Conclusion: Male sex, anemia, thrombocytopenia, febrile neutropenia and previous hospitalization in an intensive care unit increase the mortality rate in patients with cancer who were infected by P. aeruginosa.

Factores de riesgo para mortalidad en la infección por Pseudomonas aeruginosa en pacientes oncológicos hospitalizados en tres ciudades de Colombia

Introduction: Pseudomonas aeruginosa is a Gram-negative and rod-shape opportunistic bacterium that is particularly efficient in the acquisition of resistance mechanisms and its high prevalence in nosocomial infections in cancer patients. Objective: To identify risk factors for mortality in cancer patients with P. aeruginosa infection. Methodology: A descriptive study was carried out in patients with P. aeruginosa infection during the hospitalization service of “Oncólogos de Occidente” in Pereira, Armenia and Manizales during 2015. Univariate and multivariate analyzes were performed. The survival analysis was established according to the Kaplan-Meier method. A value of p<0.05 was established for it. The analyses were examined with the STATA software. This study was endorsed by the bioethics committee of the “Universidad Tecnológica de Pereira”. Results: Fourty-one patients with positive culture for P. aeruginosa were studied. Males (46.3%), anemia (46.3%), febrile neutropenia (41%), thrombocytopenia (29.3%) and previous hospitalization in an intensive care unit (29.3%) were associated with higher mortality risk (p = 0.019); a risk scale was designed with these factors (Cronbach´s alpha = 0.72). Patients who presented four of these exposures were at higher risk of mortality with a sensitivity of 68% and specificity of 90% at the moment of discharge. P. aeruginosa showed 36.6% of resistance to cefepime, 34.1% to aztreonam, the mortality rate was 26.8%. Conclusion: Male sex, anemia, thrombocytopenia, febrile neutropenia and previous hospitalization in an intensive care unit increase the mortality rate in patients with cancer who were infected by P. aeruginosa. Introducción: Pseudomonas aeruginosa es una bacteria oportunista Gram negativa particularmente eficiente en la adquisición de mecanismos de resistencia y de alta prevalencia en infecciones nosocomiales en pacientes oncológicos. Objetivo: identificar los factores de riesgo para mortalidad en pacientes oncológicos con aislamiento de P. aeruginosa. Metodología: estudio descriptivo, la población de estudio fueron los casos reportados con aislamiento de P. aeruginosa en el servicio de hospitalización de Oncólogos de Occidente en Pereira, Armenia y Manizales durante el año 2015. Se realizaron análisis univariados y multivariados; la supervivencia se estableció según el método de Kaplan-Meier. Se estableció un valor de p <0.05. Se usó el software STATA. Se tuvo aval de bioética de la Universidad Tecnológica de Pereira. Resultados: se estudió 41 casos confirmados de cultivos positivos de P. aeruginosa. El sexo masculino (46.3%), anemia (46.3%), neutropenia febril (41%), trombocitopenia (29.3%) y haber sido hospitalizado en la unidad de cuidados intensivos (29.3%) fueron asociados estadísticamente con mayor mortalidad (p=0.019); con estos resultados se diseñó una escala de riesgo (alfa de Cronbach =0.72). Los pacientes con cuatro de estas exposiciones mostraron mayor riesgo de mortalidad al egreso hospitalario con una sensibilidad del 68% y especificidad del 90%. La P. aeruginosa presentó resistencia a cefepime (36.6%) y a aztreonam (34.1%), mientras que la letalidad global fue del 26.8%. Conclusión: El sexo masculino, la coexistencia de anemia, trombocitopenia, y neutropenia febril, así como la estancia en la unidad de cuidados intensivos aumentan la mortalidad en los pacientes oncológicos infectados con P. aeruginosa

A DIGE study on the effects of salbutamol on the rat muscle proteome - an exemplar of best practice for data sharing in proteomics

BACKGROUND: Proteomic techniques allow researchers to perform detailed analyses of cellular states and many studies are published each year, which highlight large numbers of proteins quantified in different samples. However, currently few data sets make it into public databases with sufficient metadata to allow other groups to verify findings, perform data mining or integrate different data sets. The Proteomics Standards Initiative has released a series of "Minimum Information About a Proteomics Experiment" guideline documents (MIAPE modules) and accompanying data exchange formats. This article focuses on proteomic studies based on gel electrophoresis and demonstrates how the corresponding MIAPE modules can be fulfilled and data deposited in public databases, using a new experimental data set as an example. FINDINGS: We have performed a study of the effects of an anabolic agent (salbutamol) at two different time points on the protein complement of rat skeletal muscle cells, quantified by difference gel electrophoresis. In the DIGE study, a total of 31 non-redundant proteins were identified as being potentially modulated at 24 h post treatment and 110 non redundant proteins at 96 h post-treatment. Several categories of function have been highlighted as strongly enriched, providing candidate proteins for further study. We also use the study as an example of best practice for data deposition. CONCLUSIONS: We have deposited all data sets from this study in public databases for further analysis by the community. We also describe more generally how gel-based protein identification data sets can now be deposited in the PRoteomics IDEntifications database (PRIDE), using a new software tool, the PRIDESpotMapper, which we developed to work in conjunction with the PRIDE Converter application. We also demonstrate how the ProteoRed MIAPE generator tool can be used to create and share a complete and compliant set of MIAPE reports for this experiment and others

Falla cardíaca avanzada en pacientes tratados con dispositivos electrofisiológicos en una unidad de electrofisiología, Pereira (Colombia)

Revista Indexada en categoría A2 por ColcienciasLa falla cardíaca avanzada es una enfermedad prevalente con una taza alta de mortalidad y hospitalización con relación a su estadio y perfil clínico, que cuesta en Norteamérica hasta 20% de los recursos destinados para el manejo de la enfermedad cardiovascular. Se realizó un análisis transversal a una cohorte de 70 pacientes en tratamiento farmacológico y cardioresincronizador con o sin cardiodesfibirlador, o solo cardiodesfibirlador automático (ICD) implantado entre los años 2010 y 2012

COOL-LAMPS III: Discovery of a 25".9 Separation Quasar Lensed by a Merging Galaxy Cluster

In the third paper from the COOL-LAMPS Collaboration, we report the discovery of COOL J0542-2125, a gravitationally lensed quasar at $z=1.84$, observed as three images due to an intervening massive galaxy cluster at $z=0.61$. The lensed quasar images were identified in a search for lens systems in recent public optical imaging data and have separations on the sky up to 25".9, wider than any previously known lensed quasar. The galaxy cluster acting as a strong lens appears to be in the process of merging, with two sub-clusters separated by $\sim 1$ Mpc in the plane of the sky, and their central galaxies showing a radial velocity difference of $\sim 1000$ km/s. Both cluster cores show strongly lensed images of an assortment of background sources, as does the region between them. A preliminary strong lens model implies masses of $M(<250\ \rm{kpc}) = 1.79^{+0.16} _{-0.01} \times 10^{14} M_{\odot}$and$M(<250\ \rm{kpc}) = 1.48^{+0.04}_{-0.10} \times 10^{14} M_{\odot}$ for the East and West sub-clusters, respectively. This line of sight is also coincident with a ROSAT ALL-sky Survey source, centered between the two confirmed cluster halos reminiscent of other major cluster-scale mergers.Comment: 13 pages, 6 figures. Submitted to Ap

A Genome-Wide Association Study of the Protein C Anticoagulant Pathway

The Protein C anticoagulant pathway regulates blood coagulation by preventing the inadequate formation of thrombi. It has two main plasma components: protein C and protein S. Individuals with protein C or protein S deficiency present a dramatically increased incidence of thromboembolic disorders. Here, we present the results of a genome-wide association study (GWAS) for protein C and protein S plasma levels in a set of extended pedigrees from the Genetic Analysis of Idiopathic Thrombophilia (GAIT) Project. A total number of 397 individuals from 21 families were typed for 307,984 SNPs using the Infinium® 317 k Beadchip (Illumina). Protein C and protein S (free, functional and total) plasma levels were determined with biochemical assays for all participants. Association with phenotypes was investigated through variance component analysis. After correcting for multiple testing, two SNPs for protein C plasma levels (rs867186 and rs8119351) and another two for free protein S plasma levels (rs1413885 and rs1570868) remained significant on a genome-wide level, located in and around the PROCR and the DNAJC6 genomic regions respectively. No SNPs were significantly associated with functional or total protein S plasma levels, although rs1413885 from DNAJC6 showed suggestive association with the functional protein S phenotype, possibly indicating that this locus plays an important role in protein S metabolism. Our results provide evidence that PROCR and DNAJC6 might play a role in protein C and free protein S plasma levels in the population studied, warranting further investigation on the role of these loci in the etiology of venous thromboembolism and other thrombotic diseases

Measurement of the cosmic ray spectrum above 4×10184{\times}10^{18} eV using inclined events detected with the Pierre Auger Observatory

A measurement of the cosmic-ray spectrum for energies exceeding $4{\times}10^{18}$ eV is presented, which is based on the analysis of showers with zenith angles greater than $60^{\circ}$ detected with the Pierre Auger Observatory between 1 January 2004 and 31 December 2013. The measured spectrum confirms a flux suppression at the highest energies. Above $5.3{\times}10^{18}$ eV, the "ankle", the flux can be described by a power law $E^{-\gamma}$ with index $\gamma=2.70 \pm 0.02 \,\text{(stat)} \pm 0.1\,\text{(sys)}$ followed by a smooth suppression region. For the energy ($E_\text{s}$) at which the spectral flux has fallen to one-half of its extrapolated value in the absence of suppression, we find $E_\text{s}=(5.12\pm0.25\,\text{(stat)}^{+1.0}_{-1.2}\,\text{(sys)}){\times}10^{19}$ eV.Comment: Replaced with published version. Added journal reference and DO