Implication of Sphingosine-1-Phosphate Receptor 2 (S1PR2) in Differentiation and Dedifferentiation of Epithelial Renal Cells

Epithelial cell differentiation is a process that involves the mesenchymal-epithelial transition (MET) and includes cell cycle arrest, cell-cell junction maturation in addition to changes in cell migration capacity. The epithelial-mesenchymal transition (EMT) is a dynamic process by which fully differentiated epithelial cells can acquire a mesenchymal phenotype. During EMT, cell adhesion and apical-basal polarity are lost, and the cytoskeleton is reorganized. Previous results from our laboratory showed that in Madin-Darby canine kidney cells (MDCK) under different culture conditions can achieve different stages of differentiation resembling MET. Sphingosine 1-phosphate (S1P) is a bioactive sphingolipid, produced by the phosphorylation of sphingosine by sphingosine kinases (SKs), which is involved in different processes such as proliferation, cell growth, differentiation, and migration. S1P can act both intracellularly as a second messenger or extracellularly as a ligand of 5 different G protein-coupled receptors (S1PR1-5). In the present work, we evaluated the importance of S1P acting on S1PR2 in the modulation of MET and EMT. We found that there are differences in the action of S1PR2 in MDCK cells that depends on the differentiation stage. S1PR2 positively modulates the passage from polarized to differentiated cells through MET. Inhibition of S1PR2 blocks adherens junction establishment, as well as apical and basal polarity. On the other hand, once cells have acquired the differentiated phenotype, S1PR2 induces the dedifferentiation of epithelial cells through EMT. Inhibition of S1PR2 triggers changes in EMT markers, such as rearrangements of the actin cytoskeleton, expression of vimentin, and nuclear translocation of beta-catenin, as well as Slug. The expression levels of S1PR2 in the different stages of differentiation of MDCK cells did not show significant differences. Instead, immunofluorescence studies showed that during cell differentiation, S1PR2 was progressively enriched at the plasma membrane. These results suggest that the location of S1PR2 depends on the stage of cell differentiation, and this determines its role. These findings highlight the great versatility of S1P on the control of physiological and pathophysiological processes.Fil: Romero, Daniela Judith. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Santacreu, Bruno Jaime. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Tarallo, Estefania. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Favale, Nicolas Octavio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaLVI Annual Meeting Argentine Society for Biochemistry and Molecular Biology; XV Annual Meeting Argentinean Society for General MicrobiologyArgentinaSociedad Argentina de Investigación Bioquímica y Biología Molecula

Factores socio-demográficos y ambientales en pacientes con nefritis lúpica en un centro de salud de Barranquilla (2019-2020)

Las enfermedades autoinmunes (EAI), conducen a la identificación de células normales del organismo como un patógeno y se estima que alrededor de un 20% de la población sufre una EAI ; dentro de las cuales se encuentra el Lupus Eritematoso Sistémico (LES). De las complicaciones sistémicas del LES, a nivel renal, la Nefritis Lúpica (NL) es de las más frecuentes . Debido a esto, este estudio busca determinar factores sociodemográficos y ambientales en pacientes con Nefritis Lúpica en un centro de salud de Barranquilla (2019-2020). Es un estudio de corte transversal, en donde se escogieron pacientes con diagnóstico de LES, NL por biopsia renal, con o sin exposición a las variables independientes del estudio, tratados en la Clínica de la Costa, mayores de 18 años, que han comunicado en la llamada telefónica estar de acuerdo con el consentimiento informado; se excluyeron a los pacientes con desarrollo de nefritis por otra causa distinta al LES, todo aquel que tuviera antecedente de enfermedad renal previo al diagnóstico del lupus y pacientes con cualquiera otra enfermedad autoinmune. Las variables independientes son los factores socio-demográficos; como la edad, género, nivel socioeconómico, nivel educativo, y etnia; asimismo los factores ambientales de los cuales se evaluaron la exposición al humo de cigarrillo, exposición a humo de leña, exposición a luz ultravioleta, exposición a asbesto, exposición a pesticidas, exposición a sílice y exposición a sustancias químicas.PregradoMedic

Sphingosine kinase and sphingosine-1phosphate regulate epithelial cell architecture by the modulation of de novo sphingolipid synthesis

Sphingolipids regulate several aspects of cell behavior and it has been demonstrated that cells adjust their sphingolipid metabolism in response to metabolic needs. Particularly, sphingosine-1-phosphate (S1P), a final product of sphingolipid metabolism, is a potent bioactive lipid involved in the regulation of various cellular processes, including cell proliferation, cell migration, actin cytoskeletal reorganization and cell adhesion. In previous work in rat renal papillae, we showed that sphingosine kinase (SK) expression and S1P levels are developmentally regulated and control de novo sphingolipid synthesis. The aim of the present study was to evaluate the participation of SK/S1P pathway in the triggering of cell differentiation by external hypertonicity. We found that hypertonicity evoked a sharp decrease in SK expression, thus activating the de novo sphingolipid synthesis pathway. Furthermore, the inhibition of SK activity evoked a relaxation of cell-cell adherens junction (AJ) with accumulation of the AJ complex (E-cadherin/β-catenin/α-catenin) in the Golgi complex, preventing the acquisition of the differentiated cell phenotype. This phenotype alteration was a consequence of a sphingolipid misbalance with an increase in ceramide levels. Moreover, we found that SNAI1 and SNAI2 were located in the cell nucleus with impairment of cell differentiation induced by SK inhibition, a fact that is considered a biochemical marker of epithelial to mesenchymal transition. So, we suggest that the expression and activity of SK1, but not SK2, act as a control system, allowing epithelial cells to synchronize the various branches of sphingolipid metabolism for an adequate cell differentiation program.Fil: Santacreu, Bruno Jaime. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Pescio, Lucila Gisele. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Romero, Daniela Judith. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Laboratorio de Biología Celular y Molecular; ArgentinaFil: Corradi, Gerardo Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Sterin, Norma Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Favale, Nicolas Octavio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentin

Virtual Microscope, a contribution to microscopy remote learning: the tool which creates bridges between laboratory microscope analysis and new technologies

En el ámbito de las ciencias naturales, el microscopio resulta indispensable pero a la vez un desafío dada la actual situación de pandemia del COVID-19 que ha generado el cierre temporal de las instituciones de Educación Superior. Así mismo, el entrenamiento en el manejo de un microscopio, la experticia y el criterio en la observación son metas que requieren de tiempos de observación, disponibilidad de equipamiento y de la supervisión de un docente calificado. En este sentido, el uso de simuladores se presenta como una sólida estrategia para salvaguardar el sistema educativo en base a su implementación exitosa en epidemias previas. En base a estos principios, cobra relevancia el desarrollo del Microscopio Virtual, el cual consiste en un entorno digital que simula la observación e interacción a través de un microscopio, tal como se haría en el laboratorio físico. La plataforma permite a los alumnos analizar preparados obtenidos a partir de muestras reales y resolver ejercicios diseñados por los docentes, en el lugar y por el tiempo que elijan. Tomando en cuenta que nuestros alumnos son en mayoría nativos digitales, la posibilidad de aprender y adquirir experticia gracias al nexo que establece el Microscopio Virtual entre la práctica de mesada y los dispositivos móviles constituye una herramienta invaluable en la enseñanza de esta disciplina. Si bien puede concebirse a esta herramienta en primera instancia como parte de una sólida estrategia de enseñanza remota de emergencia frente a la pandemia, el Microscopio Virtual nos revela una fidedigna posibilidad que amerita su integración a la educación formal en el desarrollo de un marco híbrido de la currícula y el aprendizaje ubicuo, potenciando así el aprendizaje significativo. Reconocemos la introducción del Microscopio Virtual como parte de un nuevo ambiente que propicia una revolución significativa en la forma en que hacemos y entendemos la Educación Superior.In the field of natural sciences, the microscope is essential but at the same time a challenge considering the current pandemic situation of COVID-19 which has generated the temporary closure of Higher Education institutions. Likewise, training in the use of a microscope, experience and observation criteria are goal that require observation time, equipment availability and the supervision of a qualified teacher. In this sense, the use of simulators is presented as a solid strategy to safeguard the educational system based on its successful implementation in previous epidemics. Based on these principles, the development of the Virtual Microscope becomes more relevant, which consists of a digital environment that simulates observation and interaction through a microscope, as would be done in the physical laboratory. The platform allows students to analyze slides obtained from real samples and solve exercises designed by teachers, in the place and for the time that they choose. Taking into account that our students are mostly digital natives, the possibility of learning and acquiring expertise considering the link established by the Virtual Microscope between “laboratory microscope analysis” and mobile devices constitutes an invaluable tool in teaching this discipline. Although this tool can be conceived in the first instance as part of a solid emergency remote teaching strategy in the face of a pandemic situation, the Virtual Microscope reveals a reliable possibility that merits its integration into formal education in the development of an hybrid frame in the curriculum and ubiquitous learning, thus enhancing meaningful learning. We acknowledge the introduction of the Virtual Microscope as part of a new environment that promote a significant revolution in the way we do and understand Higher Education.Fil: Moreira Szokalo, Rocío Ayelén. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Bioquímica Clínica; ArgentinaFil: Romero, Daniela Judith. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Ciencias Biológicas. Cátedra de Biología Celular y Molecular; ArgentinaFil: Carballo, Marta Ana. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Bioquímica Clínica; ArgentinaFil: Favale, Nicolas Octavio. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Ciencias Biológicas. Cátedra de Biología Celular y Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Desarrollo de la Escala de Síntomas Internalizados en la Primera Infancia (ESIPI)

This study aims to design a scale that allows to know about emotional problems in children from two to five years. Fordoing so, a discourse analysis of educational agents and community mothers from the department of Atlántico was takenas point of departure. This is a qualitative study, which includes a discourse analysis design. The sample was constituted by35 individuals between 18 and 56 years old who belonged to medium-low socioeconomic level. Participants were dividedinto two focal groups. They answered a survey, participated in a workshop, and collaborated with three in-depth interviewsabout their experience in child development centers and community homes. The Early Childhood Internalizing SymptomScale (ESIPI) emerged as result. It includes three subscales: anxiety, depressive symptoms and somatic symptoms.El objetivo del presente estudio es diseñar una escala que alerte sobre los problemas emocionales en niños de dos a cincoaños a partir del análisis del discurso de agentes educativos y madres comunitarias, en el departamento del Atlántico. Elestudio es de corte cualitativo, e incluye diseño de análisis del discurso. Los participantes fueron 35 individuos de edadescomprendidas entre 18 y 56 años, principalmente de nivel socioeconómico medio-bajo. La muestra se dividió en dos gruposfocales, quienes contestaron una encuesta, participaron en un taller y colaboraron con tres entrevistas a profundidadsobre su experiencia en los centros de desarrollo infantil y hogares comunitarios. El resultado del estudio fue la creaciónde la Escala de Síntomas Internalizantes en Primera Infancia (ESIPI), la cual contiene tres subescalas: ansiedad, síntomasdepresivos y síntomas somáticos

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements

Design and baseline characteristics of the finerenone in reducing cardiovascular mortality and morbidity in diabetic kidney disease trial

Background: Among people with diabetes, those with kidney disease have exceptionally high rates of cardiovascular (CV) morbidity and mortality and progression of their underlying kidney disease. Finerenone is a novel, nonsteroidal, selective mineralocorticoid receptor antagonist that has shown to reduce albuminuria in type 2 diabetes (T2D) patients with chronic kidney disease (CKD) while revealing only a low risk of hyperkalemia. However, the effect of finerenone on CV and renal outcomes has not yet been investigated in long-term trials. Patients and Methods: The Finerenone in Reducing CV Mortality and Morbidity in Diabetic Kidney Disease (FIGARO-DKD) trial aims to assess the efficacy and safety of finerenone compared to placebo at reducing clinically important CV and renal outcomes in T2D patients with CKD. FIGARO-DKD is a randomized, double-blind, placebo-controlled, parallel-group, event-driven trial running in 47 countries with an expected duration of approximately 6 years. FIGARO-DKD randomized 7,437 patients with an estimated glomerular filtration rate >= 25 mL/min/1.73 m(2) and albuminuria (urinary albumin-to-creatinine ratio >= 30 to <= 5,000 mg/g). The study has at least 90% power to detect a 20% reduction in the risk of the primary outcome (overall two-sided significance level alpha = 0.05), the composite of time to first occurrence of CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure. Conclusions: FIGARO-DKD will determine whether an optimally treated cohort of T2D patients with CKD at high risk of CV and renal events will experience cardiorenal benefits with the addition of finerenone to their treatment regimen. Trial Registration: EudraCT number: 2015-000950-39; ClinicalTrials.gov identifier: NCT02545049

ARDD 2020: from aging mechanisms to interventions

Aging is emerging as a druggable target with growing interest from academia, industry and investors. New technologies such as artificial intelligence and advanced screening techniques, as well as a strong influence from the industry sector may lead to novel discoveries to treat age-related diseases. The present review summarizes presentations from the 7th Annual Aging Research and Drug Discovery (ARDD) meeting, held online on the 1st to 4th of September 2020. The meeting covered topics related to new methodologies to study aging, knowledge about basic mechanisms of longevity, latest interventional strategies to target the aging process as well as discussions about the impact of aging research on society and economy. More than 2000 participants and 65 speakers joined the meeting and we already look forward to an even larger meeting next year. Please mark your calendars for the 8th ARDD meeting that is scheduled for the 31st of August to 3rd of September, 2021, at Columbia University, USA

Identification of genetic variants associated with Huntington's disease progression: a genome-wide association study

Background Huntington's disease is caused by a CAG repeat expansion in the huntingtin gene, HTT. Age at onset has been used as a quantitative phenotype in genetic analysis looking for Huntington's disease modifiers, but is hard to define and not always available. Therefore, we aimed to generate a novel measure of disease progression and to identify genetic markers associated with this progression measure. Methods We generated a progression score on the basis of principal component analysis of prospectively acquired longitudinal changes in motor, cognitive, and imaging measures in the 218 indivduals in the TRACK-HD cohort of Huntington's disease gene mutation carriers (data collected 2008–11). We generated a parallel progression score using data from 1773 previously genotyped participants from the European Huntington's Disease Network REGISTRY study of Huntington's disease mutation carriers (data collected 2003–13). We did a genome-wide association analyses in terms of progression for 216 TRACK-HD participants and 1773 REGISTRY participants, then a meta-analysis of these results was undertaken. Findings Longitudinal motor, cognitive, and imaging scores were correlated with each other in TRACK-HD participants, justifying use of a single, cross-domain measure of disease progression in both studies. The TRACK-HD and REGISTRY progression measures were correlated with each other (r=0·674), and with age at onset (TRACK-HD, r=0·315; REGISTRY, r=0·234). The meta-analysis of progression in TRACK-HD and REGISTRY gave a genome-wide significant signal (p=1·12 × 10−10) on chromosome 5 spanning three genes: MSH3, DHFR, and MTRNR2L2. The genes in this locus were associated with progression in TRACK-HD (MSH3 p=2·94 × 10−8 DHFR p=8·37 × 10−7 MTRNR2L2 p=2·15 × 10−9) and to a lesser extent in REGISTRY (MSH3 p=9·36 × 10−4 DHFR p=8·45 × 10−4 MTRNR2L2 p=1·20 × 10−3). The lead single nucleotide polymorphism (SNP) in TRACK-HD (rs557874766) was genome-wide significant in the meta-analysis (p=1·58 × 10−8), and encodes an aminoacid change (Pro67Ala) in MSH3. In TRACK-HD, each copy of the minor allele at this SNP was associated with a 0·4 units per year (95% CI 0·16–0·66) reduction in the rate of change of the Unified Huntington's Disease Rating Scale (UHDRS) Total Motor Score, and a reduction of 0·12 units per year (95% CI 0·06–0·18) in the rate of change of UHDRS Total Functional Capacity score. These associations remained significant after adjusting for age of onset. Interpretation The multidomain progression measure in TRACK-HD was associated with a functional variant that was genome-wide significant in our meta-analysis. The association in only 216 participants implies that the progression measure is a sensitive reflection of disease burden, that the effect size at this locus is large, or both. Knockout of Msh3 reduces somatic expansion in Huntington's disease mouse models, suggesting this mechanism as an area for future therapeutic investigation

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030