Molecular characterization of TC964, a novel antigenic protein from trypanosoma cruzi

The Tc964 protein was initially identified by its presence in the interactome associated with the LYT1 mRNAs, which code for a virulence factor of Trypanosoma cruzi. Tc964 is annotated in the T. cruzi genome as a hypothetical protein. According to phylogenetic analysis, the protein is conserved in the different genera of the Trypanosomatidae family; however, recognizable orthologues were not identified in other groups of organisms. Therefore, as a first step, an in-depth molecular characterization of the Tc946 protein was carried out. Based on structural predictions and molecular dynamics studies, the Tc964 protein would belong to a particular class of GTPases. Subcellular fractionation analysis indicated that Tc964 is a nucleocytoplasmic protein. Additionally, the protein was expressed as a recombinant protein in order to analyze its antigenicity with sera from Chagas disease (CD) patients. Tc964 was found to be antigenic, and B-cell epitopes were mapped by the use of synthetic peptides. In parallel, the Leishmania major homologue (Lm964) was also expressed as recombinant protein and used for a preliminary evaluation of antigen cross-reactivity in CD patients. Interestingly, Tc964 was recognized by sera from Chronic CD (CCD) patients at different stages of disease severity, but no reactivity against this protein was observed when sera from Colombian patients with cutaneous leishmaniasis were analyzed. Therefore, Tc964 would be adequate for CD diagnosis in areas where both infections (CD and leishmaniasis) coexist, even though additional assays using larger collections of sera are needed in order to confirm its usefulness for differential serodiagnosisThis research was funded by Ministerio de Ciencia,Tecnología e Innovación (Minciencias) and Pontificia Universidad Javeriana, research project ID PPTA 120356933228 granted to C.J.P. The article publication was funded by the Vicerrectoría de Investigación from the Pontificia Universidad Javeriana, code 120813F0401200. The Network of Tropical Diseases Research RICET (RD16/0027/0008, Instituto de Salud Carlos III and co-funded by FEDER) to J.M.R., and grants from the Spanish Ministerio de Ciencia, Innovación y Universidades/Agencia Estatal de Investigación RTC-2017-6494-1 and RTI2018-094434-B-I00 (MCIU/AEI/FEDER, UE) to P.G.-P., E.R.-M and E.R.R. were supported by Minciencias convocatoria doctorados nacionales 647-2014 and convocatoria jóvenes investigadores e innovadores 706-2015, respectivel

LRP10, PGK1 and RPLP0: best reference genes in periprostatic adipose tissue under obesity and prostate cancer conditions

Obesity (OB) is a metabolic disorder characterized by adipose tissue dysfunction that has emerged as a health problem of epidemic proportions in recent decades. OB is associated with multiple comorbidities, including some types of cancers. Specifically, prostate cancer (PCa) has been postulated as one of the tumors that could have a causal relationship with OB. Particularly, a specialized adipose tissue (AT) depot known as periprostatic adipose tissue (PPAT) has gained increasing attention over the last few years as it could be a key player in the pathophysiological interaction between PCa and OB. However, to date, no studies have defined the most appropriate internal reference genes (IRGs) to be used in gene expression studies in this AT depot. In this work, two independent cohorts of PPAT samples (n = 20/n = 48) were used to assess the validity of a battery of 15 literature-selected IRGs using two widely used techniques (reverse transcription quantitative PCR [RT-qPCR] and microfluidic-based qPCR array). For this purpose, ΔCt method, GeNorm (v3.5), BestKeeper (v1.0), NormFinder (v.20.0), and RefFinder software were employed to assess the overall trends of our analyses. LRP10, PGK1, and RPLP0 were identified as the best IRGs to be used for gene expression studies in human PPATs, specifically when considering PCa and OB conditions

Reduced L-carnitine transport in aortic endothelial cells from spontaneously hypertensive rats

Impaired L-carnitine uptake correlates with higher blood pressure in adult men, and L-carnitine restores endothelial function in aortic rings from spontaneously hypertensive rat (SHR). Thus, endothelial dysfunction in hypertension could result from lower L-carnitine transport in this cell type. L-Carnitine transport is mainly mediated by novel organic cation transporters 1 (Octn1, Na+-independent) and 2 (Octn2, Na+-dependent); however, their kinetic properties and potential consequences in hypertension are unknown. We hypothesize that L-carnitine transport kinetic properties will be altered in aortic endothelium from spontaneously hypertensive rats (SHR). L-Carnitine transport was measured at different extracellular pH (pHo 5.5–8.5) in the absence or presence of sodium in rat aortic endothelial cells (RAECs) from non-hypertensive Wistar-Kyoto (WKY) rats and SHR. Octn1 and Octn2 mRNA relative expression was also determined. Dilation of endothelium-intact or denuded aortic rings in response to calcitonine gene related peptide (CGRP, 0.1–100 nmol/L) was measured (myography) in the absence or presence of L-carnitine. Total L-carnitine transport was lower in cells from SHR compared with WKY rats, an effect due to reduced Na+-dependent (Na+dep) compared with Na+-independent (Na+indep) transport components. Saturable L-carnitine transport kinetics show maximal velocity (Vmax), without changes in apparent Km for Na+indep transport in SHR compared with WKY rats. Total and Na+dep component of transport were increased, but Na+indep transport was reduced by extracellular alkalization in WKY rats. However, alkalization reduced total and Na+indep transport in cells from SHR. Octn2 mRNA was higher than Octn-1 mRNA expression in cells from both conditions. Dilation of artery rings in response to CGRP was reduced in vessels from SHR compared with WKY rats. CGRP effect was endothelium-dependent and restored by L-carnitine. All together these results suggest that reduced L-carnitine transport (likely via Na+-dependent Octn2) could limit this compound's potential beneficial effects in RAECs from SHR

Adipose tissue depot-specific intracellular and extracellular cues contributing to insulin resistance in obese individuals.

Adipose tissue dysregulation in obesity strongly influences systemic metabolic homeostasis and is often linked to insulin resistance (IR). However, the molecular mechanisms underlying adipose tissue dysfunction in obesity are not fully understood. Herein, a proteomic analysis of subcutaneous (SC) and omental (OM) fat from lean subjects and obese individuals with different degrees of insulin sensitivity was performed to identify adipose tissue biomarkers related to obesity-associated metabolic disease. Our results suggest that dysregulation of both adipose tissue extracellular matrix (ECM) organization and intracellular trafficking processes may be associated with IR in obesity. Thus, abnormal accumulation of the small leucine-rich proteoglycan, lumican, as observed in SC fat of IR obese individuals, modifies collagen I organization, impairs adipogenesis and activates stress processes [endoplasmic reticulum and oxidative stress] in adipocytes. In OM fat, IR is associated with increased levels of the negative regulator of the Rab family of small GTPases, GDI2, which alters lipid storage in adipocytes by inhibiting insulin-stimulated binding of the Rab protein, Rab18, to lipid droplets. Together, these results indicate that lumican and GDI2 might play depot-dependent, pathogenic roles in obesity-associated IR. Our findings provide novel insights into the differential maladaptive responses of SC and OM adipose tissue linking obesity to IR

The caveolae‐associated coiled‐coil protein, NECC2, regulates insulin signalling in Adipocytes

Adipocyte dysfunction in obesity is commonly associated with impaired insulin signalling in adipocytes and insulin resistance. Insulin signalling has been associated with caveolae, which are coated by large complexes of caveolin and cavin proteins, along with proteins with membrane‐binding and remodelling properties. Here, we analysed the regulation and function of a component of caveolae involved in growth factor signalling in neuroendocrine cells, neuroendocrine long coiled‐coil protein‐2 (NECC2), in adipocytes. Studies in 3T3‐L1 cells showed that NECC2 expression increased during adipogenesis. Furthermore, NECC2 co‐immunoprecipitated with caveolin‐1 (CAV1) and exhibited a distribution pattern similar to that of the components of adipocyte caveolae, CAV1, Cavin1, the insulin receptor and cortical actin. Interestingly, NECC2 overexpression enhanced insulin‐activated Akt phosphorylation, whereas NECC2 downregulation impaired insulin‐induced phosphorylation of Akt and ERK2. Finally, an up‐regulation of NECC2 in subcutaneous and omental adipose tissue was found in association with human obesity and insulin resistance. This effect was also observed in 3T3‐L1 adipocytes exposed to hyperglycaemia/hyperinsulinemia. Overall, the present study identifies NECC2 as a component of adipocyte caveolae that is regulated in response to obesity and associated metabolic complications, and supports the contribution of this protein as a molecular scaffold modulating insulin signal transduction at these membrane microdomains.La disfunción de los adipocitos en la obesidad se asocia comúnmente con la alteración de la señalización de la insulina en los adipocitos y la resistencia a la insulina. La señalización de la insulina se ha asociado con las caveolas, que están recubiertas por grandes complejos de proteínas de caveolina y cavina, junto con proteínas con propiedades de remodelación y unión a la membrana. Aquí, analizamos la regulación y la función de un componente de las caveolas involucrado en la señalización del factor de crecimiento en las células neuroendocrinas, la proteína 2 neuroendocrina de espiral larga (NECC 2 ) , en los adipocitos. Los estudios en células 3T3‐L1 mostraron que la expresión de NECC 2 aumentó durante la adipogénesis. Además, NECC 2 co‐inmunoprecipitado con caveolina‐1 ( CAV1) y mostró un patrón de distribución similar al de los componentes de las caveolas adipocitarias, CAV 1, Cavin1, el receptor de insulina y la actina cortical. Curiosamente, la sobreexpresión de NECC 2 mejoró la fosforilación de Akt activada por insulina, mientras que la regulación negativa de NECC 2 perjudicó la fosforilación de Akt y ERK 2 inducida por insulina . resistencia a la insulina. Este efecto también se observó en adipocitos 3T3‐L1 expuestos a hiperglucemia/hiperinsulinemia. En general, el presente estudio identifica NECC2 como un componente de las caveolas de los adipocitos que se regula en respuesta a la obesidad y las complicaciones metabólicas asociadas, y respalda la contribución de esta proteína como un andamio molecular que modula la transducción de señales de insulina en estos microdominios de membrana

Cerebro arte y creatividad

En el seminario Cerebro, Arte y Creatividad, realizado en mayo del 2000, en la Universidad Nacional, cuyas conferencias hemos integrado en este libro, hemos querido aproximamos a diversas manifestaciones del arte, diversas variedades de creatividad, desde la perspectiva de las funciones cerebrales y desde la perspectiva de los principios psicológicos que gobiernan la actividad creativa. Se explora cómo los hallazgos relativos al funcionamiento y especialización funcional de los hemisferios cerebrales inciden en las manifestaciones artísticas y en nuestra percepción estética. A su vez, se ilustran interpretaciones de las producciones artísticas, desde la aproximación psicoanalítica. Se presenta una multiplicidad de aproximaciones hacia la comprensión y exploración de las relaciones entre cerebro, arte y creatividad. Como se verá, hay muchos misterios por explorar. Pero Einstein decía: "lo más bello que podemos experimentar, es el misterio. Es la fuente del verdadero arte y de la verdadera ciencia". Cada día será más fascinante ir más lejos, con nuevas preguntas y nuevos misterios. La portada de este libro, fue seleccionada entre aquellas enviadas al concurso para el afiche del simposio Cerebro, Arte y Creatividad. / Contenido. Preliminares; Capítulo 1 - Neuroanatomía y psicología de la percepción estética; Capítulo 2 - Cerebro y pintura; Capítulo 3 - Cerebro y música; Capítulo 4 - Cerebro y literatura; Capítulo 5 - La radiología en el arte; Capítulo 6 - Psicoanálisis, arte y creatividad; Anexos

Design and characterization of an ocular topical liposomal preparation to replenish the lipids of the tear film

Purpose: Dry eye (DE) includes a group of diseases related to tear film disorders. Current trends for DE therapy focus on providing lipid components to replace the damaged lipid layer. Formulations that contain aqueous and mucin like compounds may have additional therapeutic benefits for DE patients. The aim of this work was to design and evaluate novel formulations having the potential to become topical treatment for DE. Methods: Unpreserved liposomal formulations composed of phosphatidylcholine (PC), cholesterol, and α tocopherol (vit E) were prepared by the thin-film hydration technique. Formulations were characterized in terms of liposome size, pH, surface tension, osmolarity, and viscosity. In vitro tolerance assays were performed on macrophage, human corneal and conjunctival cell lines at short and long term exposures. In vivo ocular tolerance was studied after instillation of the formulation. Results: The mean liposome size was less than 1 μm and surface tension <30 50 mN/m for all formulations. The final liposomal formulation (PC:cholesterol:vit E in a ratio 8:1:0.8) had physiological values of pH (6.45 ± 0.09), osmolarity (289.43 ± 3.28 mOsm), and viscosity (1.82 ± 0.02 mPa·s). Cell viability was greater than 80% in the corneal and conjunctival cells. This formulation was well tolerated by experimental animals. Conclusions: The unpreserved liposomal formulation has suitable properties to be administered by topical ophthalmic route. The liposome-based artificial tear had good in vitro and in vivo tolerance responses. This formulation composed of a combination of liposomes and bioadhesive polymers may be employed successfully as a tear film substitute in DE therapy

Brucella Genetic Variability in Wildlife Marine Mammals Populations Relates to Host Preference and Ocean Distribution

Intracellular bacterial pathogens probably arose when their ancestor adapted from a free-living environment to an intracellular one, leading to clonal bacteria with smaller genomes and less sources of genetic plasticity. Still, this plasticity is needed to respond to the challenges posed by the host. Members of the Brucella genus are facultative-extracellular intracellular bacteria responsible for causing brucellosis in a variety of mammals. The various species keep different host preferences, virulence, and zoonotic potential despite having 97–99% similarity at genome level. Here, we describe elements of genetic variation in Brucella ceti isolated from wildlife dolphins inhabiting the Pacific Ocean and the Mediterranean Sea. Comparison with isolates obtained from marine mammals from the Atlantic Ocean and the broader Brucella genus showed distinctive traits according to oceanic distribution and preferred host. Marine mammal isolates display genetic variability, represented by an important number of IS711 elements as well as specific IS711 and SNPs genomic distribution clustering patterns. Extensive pseudogenization was found among isolates from marine mammals as compared with terrestrial ones, causing degradation in pathways related to energy, transport of metabolites, and regulation/ transcription. Brucella ceti isolates infecting particularly dolphin hosts, showed further degradation of metabolite transport pathways as well as pathways related to cell wall/membrane/envelope biogenesis and motility. Thus, gene loss through pseudogenization is a source of genetic variation in Brucella, whichinturn, relates to adaptation to different hosts.This is relevant to understand the natural history of bacterial diseases, their zoonotic potential, and the impact of human interventions such as domestication.Comisión Nacional para la Gestión de la Biodiversidad/[R-028-203-OT]/CONAGEBIO/Costa RicaMinisterio de Ciencia, Tecnología y Telecomunicaciones/[FV-004-13]/MICITT/Costa RicaWellcome Trust/[098051]/WT/LondresWellcome Trust/[106690/Z/14/Z]/WT/LondresUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Centro de Investigación en Enfermedades Tropicales (CIET

Marco activo de recursos de innovación docente: Madrid

Una guía de espacios e instituciones para actividades educativas complementarias en enseñanza secundaria y Formación Profesional