Cristina Maldonado - Academia.edu (original) (raw)

Papers by Cristina Maldonado

PLOS ONE, 2020

Allergic asthma is the most common phenotype of the pathology, having an early-onset in childhood... more Allergic asthma is the most common phenotype of the pathology, having an early-onset in childhood and producing a Th2-driven airways remodeling process that leads to symptoms and pathophysiological changes. The avoidance of aeroallergen exposure in early life has been shown to prevent asthma, but without repeated success and with the underlying preventive mechanisms at the beginning of asthma far to be fully recognized. In the present study, we aimed to evaluate if neonatal LPS-induced boost in epithelial host defenses contribute to prevent OVA-induced asthma in adult mice. To this, we focused on the response of bronchiolar club cells (CC), which are highly specialized in maintaining the epithelial homeostasis in the lung. In these cells, neonatal LPS administration increased the expression of TLR4 and TNFα, as well as the immunodulatory/antiallergic proteins: club cell secretory protein (CCSP) and surfactant protein D (SP-D). LPS also prevented mucous metaplasia of club cells and reduced the epidermal growth factor receptor (EGFR)-dependent mucin overproduction, with mice displaying normal breathing patterns after OVA challenge. Furthermore, the overexpression of the epithelial Th2-related molecule TSLP was blunted, and normal TSLP and IL-4 levels were found in the bronchoalveolar lavage. A lower eosinophilia was detected in LPS-pretreated mice, along with an increase in phagocytes and regulatory cells (CD4+CD25+FOXP3+ and CD4+IL-10+), together with higher levels of IL-12 and TNFα. In conclusion, our study demonstrates stable asthma-preventive epithelial effects promoted by neonatal LPS stimulation, leading to the presence of regulatory cells in the lung. These anti-allergic dynamic mechanisms would be overlaid in the epithelium, favored by an adequate epidemiological environment, during the development of asthma.

Journal of Cellular Physiology, 2017

Prostatic smooth muscle cells (pSMCs) differentiation is a key factor for prostatic homeostasis, ... more Prostatic smooth muscle cells (pSMCs) differentiation is a key factor for prostatic homeostasis, with androgens exerting multiple effects on these cells. Here, we demonstrated that the myodifferentiator complex Srf/Myocd is up-regulated by testosterone in a dose-dependent manner in primary cultures of rat pSMCs, which was associated to the increase in Acta2, Cnn1, and Lmod1 expressions. Blocking Srf or Myocd by siRNAs inhibited the myodifferentiator effect of testosterone. While LPS led to a dedifferentiated phenotype in pSMCs, characterized by down-regulation of Srf/Myocd and smooth muscle cell (SMC)-restricted genes, endotoxin treatment on Myocd-overexpressing cells did not result in phenotypic alterations. Testosterone at a physiological dose was able to restore the muscular phenotype by normalizing Srf/Myocd expression in inflammation-induced dedifferentiated pSMCs. Moreover, the androgen reestablished the proliferation rate and IL-6 secretion increased by LPS. These results provide novel evidence regarding the myodifferentiating role of testosterone on SMCs by modulating Srf/Myocd. Thus, androgens preserve prostatic SMC phenotype, which is essential to maintain the normal structure and function of the prostate. This article is protected by copyright. All rights reserved.

La Enfermedad Pulmonar Obstructiva Cronica (EPOC) ocupa la sexta posicion entre las causas de mue... more La Enfermedad Pulmonar Obstructiva Cronica (EPOC) ocupa la sexta posicion entre las causas de muerte en todo el mundo, siendo ademas una importante causa de incapacidad, que se ve reflejada en perdidas de la productividad y en altos costos economicos. Por lo tanto, son prioritarias las politicas sanitarias orientadas a frenar el crecimiento y el indice de morbi-mortalidad de la EPOC y de las enfermedades pulmonares en general. La epidemia actual de la EPOC es debida en parte al habito de fumar; sin embargo, solo el 20% de los fumadores la desarrollan, desconociendose, entre otros procesos fisiopatogenicos, las bases celulares y moleculares que determinan que un fumador desarrolle la enfermedad. Este proyecto resulta de la interaccion de profesionales del area clinica, neumonologos del SANATORIO ALLENDE, y del area basica que se desempenan como docentes investigadores en la FACULTAD DE CIENCIAS MEDICAS de la UNC. Nuestro OBJETIVO GENERAL es investigar aspectos aun no evaluados sobre la EPOC, en la busqueda de nuevos parametros adecuados para medir del grado de inflamacion bronquial, que permitan mejorar su prevencion, diagnostico y tratamiento. Una herramienta fundamentalmente usada es la inflamometria del esputo inducido, resultando sencillo, no invasivo y de bajo costo, puesto a punto por primera vez en Argentina por integrantes de esta red. Los objetivos especificos estan orientados a responder diferentes hipotesis planteadas a partir de la problematica diaria del consultorio y desde conocimientos recientes sobre las celulas inflamatorias. El primer aspecto de interes es dilucidar la incidencia de la NETosis, muerte alternativa de los neutrofilos, en el mantenimiento del proceso inflamatorio y de los sintomas propios de la EPOC. Este es un tipo de muerte en el cual se libera el DNA junto al contenido intragranular como mecanismo antimicrobiano y que, segun datos preliminares de nuestro grupo, estaria descontrolado en pacientes con EPOC [18], exponiendo DNA y antigenos propios. De alli nuestros objetivos tendientes en primer lugar a identificar y corroborar NETosis en esputo inducido mediante microscopia confocal y electronica, asociandola con la severidad en la sintomatologia de la EPOC. En segundo lugar se determinara la relacion de NETosis con posibles manifestaciones autoinmunes en la EPOC, para lo cual se determinaran anticuerpos sericos anti-DNA y anticuerpos anti-proteinas del neutrofilo por ELISA. Un tercer objetivo estara orientado mas directamente a analizar la correlacion entre NETosis y el grado de inflamacion de la via aerea, traducido en la relacion entre macrofagos proinflamatorios (M1) y antiinflamatorios (M2) presentes en el esputo inducido, que sera examinada mediante citometria de flujo. Una segunda incognita a abordar proviene de un clasico interrogante en la clinica neumonologica acerca de una posible entidad comun entre la EPOC y el asma. En este contexto, muchos de los pacientes con EPOC tendrian en realidad un fondo asmatico, en el cual el habito tabaquico desencadenaria el proceso inflamatorio. Esta hipotesis se refleja en nuestro ultimo objetivo especifico, investigando marcadores de asma en pacientes con EPOC, a fin de evidenciar un posible solapamiento entre ambas entidades y reorientar la terapia en estos pacientes. En el planteo y ejecucion de estos objetivos, se incluiran comparativamente pacientes con EPOC, fumadores sanos, asmaticos de desarrollo adulto y voluntarios sanos. El desarrollo del presente proyecto impactara significativamente no solo en la identificacion de nuevos blancos terapeuticos que optimicen la resolucion de las enfermedades pulmonares, sino tambien en la formacion de recursos humanos, fortaleciendo vinculos entre los hospitales, los laboratorios de investigacion y las casas de altos estudios de nuestra provincia.

Experimental Biology and Medicine, 2014

Atopic asthma is a chronic allergic disease that involves T-helper type 2 (Th2)-inflammation and ... more Atopic asthma is a chronic allergic disease that involves T-helper type 2 (Th2)-inflammation and airway remodeling. Bronchiolar club cells (CC) and alveolar macrophages (AM) are sentinel cells of airway barrier against inhaled injuries, where allergy induces mucous metaplasia of CC and the alternative activation of AM, which compromise host defense mechanisms and amplify Th2-inflammation. As there is evidence that high levels of environmental endotoxin modulates asthma, the goal of this study was to evaluate if the activation of local host defenses by Lipopolysaccharide (LPS) previous to allergy development can contribute to preserving CC and AM protective phenotypes. Endotoxin stimulus before allergen exposition reduced hallmarks of allergic inflammation including eosinophil influx, Interleukin-4 and airway hyperreactivity, while the T-helper type 1 related cytokines IL-12 and Interferon-γ were enhanced. This response was accompanied by the preservation of the normal CC phenotype a...

Veterinary Ophthalmology, 2009

To describe the functional and structural characteristics of the cornea in healthy Guinea pigs. H... more To describe the functional and structural characteristics of the cornea in healthy Guinea pigs. Healthy male and female pigmented and albino Guinea pigs (Caviaporcellus) aged 3-5 months old were used. The animals' corneas underwent different in vivo studies including: slit-lamp biomicroscopy, fluorescein staining (FS), break-up time test (BUT), confocal microscopy and pachymetry. The corneas were also studied histopathologically with light microscopy, immunohistochemistry and transmission electron microscopy. No significant differences were found between pigmented and albino animals, male and female, OD and OS in any study performed. The differences on corneal thickness values were not significant among central (227.85 +/- 14.09 microm) and upper and temporal peripheral regions (226.60 +/- 12.50 and 225.70 +/- 14.40 microm, respectively). All histological studies performed permitted identification and precise description of the different corneal structures in Guinea pigs: the stratified epithelium (45.52 +/- 5.26 microm), Bowman's layer (2.23 +/- 0.38 microm), stroma (163.69 +/- 4.90 microm), Descemet's membrane (3.96 +/- 0.46 microm) and the endothelium (5.09 +/- 0.71 microm). Combining results from all eyes mean and SD from corneal BUT values was 4.98 +/- 1.67 s. Corneas often showed discrete superficial erosions being the FS positive in both eyes from all the animals. This study provides a detailed in vivo and postfixed histological description of the Guinea pig's cornea and information about the physiological tests.

International Journal of Experimental Pathology, 2013

Histochemistry and Cell Biology, 2008

The objectives of the present work were to assess whether epithelial cells from the diVerent segm... more The objectives of the present work were to assess whether epithelial cells from the diVerent segments of epididymis express TR 1-1 isoforms, to depict its subcellular immunolocalization and to evaluate changes in their expression in rats experimentally submitted to a hypothyroid state by injection of 131I. In euthyroid and hypothyroid groups, TR protein was expressed in epididymal epithelial cells, mainly in the cytoplasmic compartment while only a few one showed a staining in the nucleus as well. A similar TR immunostaining pattern was detected in the diVerent segments of the epididymis. In hypothyroid rats, the number of TR-immunoreactive epithelial cells as well as the intensity of the cytoplasmic staining signiWcantly increased in all sections analyzed. In consonance to the immunocytochemical analysis, the expression of TR 1-1 isoforms, assessed by Western blot revealed signiWcantly higher levels of TR in cytosol compared to the nuclear fractions. Furthermore, TR expression of both 1 and 1 isoforms and their mRNA levels were increased by the hypothyroid state. The immuno-electron-microscopy showed speciWc reaction for TR in principal cells associated with eucromatin, cytosolic matrix and mitochondria. The diVerences in expression levels assessed in control and thyroidectomized rats ascertain a speciWc function of TH on this organ.

… and Cell Biology, 2008

Histochem Cell Biol (2008) 129:631–642 DOI 10.1007/s00418-008-0397-8 ... Thyroid hormone receptor... more Histochem Cell Biol (2008) 129:631–642 DOI 10.1007/s00418-008-0397-8 ... Thyroid hormone receptor 1 – 1 expression in epididymal ... Ana Lucía De Paul · Jorge Humberto Mukdsi · Claudia Gabriela Pellizas · María Montesinos · Silvina Gutiérrez · Sebastián Susperreguy · Alberto ...

Applications of Immunocytochemistry, 2012

Applications of Immunocytochemistry 66 appropriate selection of an embedding resin and the ready ... more Applications of Immunocytochemistry 66 appropriate selection of an embedding resin and the ready availability of the specific antibodies for the molecules whose ultrastructural location needs to be determined.

A pesar de los avances fisiopatogenicos y terapeuticos, el asma permanece sin control. El objetiv... more A pesar de los avances fisiopatogenicos y terapeuticos, el asma permanece sin control. El objetivo de este ensayo es mostrar evidencias clinicas-cientificas que los instrumentos utilizados en el manejo del asma no son herramientas suficientes y proponer una estrategia para el abordaje y su control en forma objetiva, teniendo como blanco terapeutico la inflamacion de la via aerea segun parametros celulares del esputo inducido. Evaluamos en 15 pacientes asmaticos bajo tratamiento: el grado de control del asma (GCA) en base a criterios clinicos, espirometricos y el cuestionario de control, segun recomendaciones de GINA (Global Initiative for Asthma). Posteriormente todos los pacientes fueron sometidos a la recoleccion de una muestra de esputo inducido (EI) para realizar el recuento diferencial de celulas segun tecnica previamente validada. Del total estudiados, obtuvimos que 7 pacientes, a pesar que se encontraban totalmente controlados segun parametros de GCA 5 presentaban un perfil i...

Revista de la Facultad de Ciencias Médicas de Córdoba

Objetivo: Evaluar la prevalencia de riesgo de asma en adolescentes expuestos a ceniza volcánica a... more Objetivo: Evaluar la prevalencia de riesgo de asma en adolescentes expuestos a ceniza volcánica a un año de la erupción del Volcán Calbuco, en 2 ciudades con diferentes grado de afectación. Métodologia: Estudio de Corte Transversal, en adolescentes de 13-14 años en dos ciudades con diferente grado de exposición a ceniza volcánica: San Carlos de Bariloche y Cipolletti. Se aplicó cuestionario y video-cuestionario ISAAC para determinar riesgo de asma. Las variables demográficas y percepción de exposición, se obtuvieron por cuestionario previo a la aplicación de metodología ISAAC. Resultados: Se encuestaron 511 sujetos en total. Se reportó prevalencia de síntomas de riesgo de asma en Bariloche resultó de 14% comparado con Cipolletti que reportó 10% (p=0.32). Se informó mayor exposición a ceniza volcánica en la ciudad de Bariloche vs. Cipolletti; 14% y 6% respectivamente con p<0.05. Conclusiones:La falta diferencia estadisticamente significativa respecto a la prevalencia de sintomas d...

Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology

Glutamine (GLN) avoids the inhibition of the intestinal Ca2+ absorption caused by menadione (MEN)... more Glutamine (GLN) avoids the inhibition of the intestinal Ca2+ absorption caused by menadione (MEN) through oxidative stress. The purpose of this study was to elucidate whether molecules of transcellular and/or paracellular pathways of intestinal Ca2+ absorption are involved in the GLN action and underlying mechanisms. One-month old chicks were divided in four groups: 1) controls, 2) MEN treated, 3) GLN treated and 4) GLN + MEN treated. The morphology of intestinal villi, the intestinal Ca2+ absorption and the molecules involved in the transcellular and paracellular pathways were analyzed. Markers of autophagy and inflammation were also evaluated. The data demonstrated that GLN protected both transcellular and paracellular pathways. GLN avoided morphological changes in the intestine caused by MEN. GLN protected the gene expression of transporters involved in the transcellular pathway and the gene and protein expression of molecules belonging to the paracellular pathways altered by MEN. GLN increased the LC3-II protein expression and the number of acidic vesicular organelles, markers of autophagy, and blocked an increase in the NFkB protein expression in the nuclei and in the IL-6 gene expression caused by MEN. In conclusion, GLN protects both transcellular and paracellular pathways of intestinal Ca2+ absorption by increasing autophagy and blocking inflammation.

Frontiers in immunology, 2018

Neutrophils are major effectors of acute inflammation against infection and tissue damage, with a... more Neutrophils are major effectors of acute inflammation against infection and tissue damage, with ability to adapt their phenotype according to the microenvironment. Although sex hormones regulate adaptive immune cells, which explains sex differences in immunity and infection, little information is available about the effects of androgens on neutrophils. We therefore aimed to examine neutrophil recruitment and plasticity in androgen-dependent and -independent sites under androgen manipulation. By using a bacterial model of prostate inflammation, we showed that neutrophil recruitment was higher in testosterone-treated rats, with neutrophil accumulation being positively correlated to serum levels of testosterone and associated to stronger inflammatory signs and tissue damage. Testosterone also promoted LPS-induced neutrophil recruitment to the prostate, peritoneum, and liver sinusoids, as revealed by histopathology, flow cytometry, and intravital microscopy. Strikingly, neutrophils in p...

Frontiers in immunology, 2018

Galectins, a family of animal lectins characterized by their affinity for N-acetyllactosamine-enr... more Galectins, a family of animal lectins characterized by their affinity for N-acetyllactosamine-enriched glycoconjugates, modulate several immune cell processes shaping the course of innate and adaptive immune responses. Through interaction with a wide range of glycosylated receptors bearing complex branched N-glycans and core 2-O-glycans, these endogenous lectins trigger distinct signaling programs thereby controling immune cell activation, differentiation, recruitment and survival. Given the unique features of mucosal inflammation and the differential expression of galectins throughout the gastrointestinal tract, we discuss here key findings on the role of galectins in intestinal inflammation, particularly Crohn's disease, ulcerative colitis, and celiac disease (CeD) patients, as well as in murine models resembling these inflammatory conditions. In addition, we present new data highlighting the regulated expression of galectin-1 (Gal-1), a proto-type member of the galectin famil...

Molecular cancer research : MCR, Jan 9, 2018

Emerging evidence suggests that unregulated Toll-like receptors (TLRs) signaling promotes tumor s... more Emerging evidence suggests that unregulated Toll-like receptors (TLRs) signaling promotes tumor survival signals, thus favoring tumor progression. Here, the mechanism underlying TLR4 overexpression in papillary thyroid carcinomas (PTCs) mainly harboring the BRAFV600E mutation was studied. TLR4 was overexpressed in PTCs compared to non-neoplastic thyroid tissue. Moreover, paired clinical specimens of primary PTC and its lymph node metastasis showed a significant upregulation of TLR4 levels in the metastatic tissues. In agreement, conditional BRAFV600E expression in normal rat thyroid cells and mouse thyroid tissue upregulated TLR4 expression levels. Furthermore, functional TLR4 expression was demonstrated in PTC cells by increased NF-κB transcriptional activity in response to the exogenous TLR4-agonist lipopolysaccharide (LPS). Of note, The Cancer Genome Atlas (TCGA) data analysis revealed that BRAFV600E-positive tumors with high TLR4 expression were associated with shorter disease-f...

Cell biology international, Jan 28, 2017

The prostate gland is a strictly androgen-dependent organ which is also the main target of infect... more The prostate gland is a strictly androgen-dependent organ which is also the main target of infectious and inflammatory diseases in the male reproductive tract. Host defenses and immunity of the gland have unique features to maintain a constant balance between response and tolerance to diverse antigens. In this context, the effects of reproductive hormones on the male tract are thus complex and have just started to be defined. From the classical description of "the prostatic antibacterial factor," many host defense proteins with potent microbicidal and anti-tumoral activities have been described in the organ. Indeed, it has been proposed a central role for resident cells, that is, epithelial and smooth muscle cells, in the prostatic response against injuries. However, these cells also represent the target of the inflammatory damage, leading to the development of a Proliferative Inflammatory Atrophy-like process in the epithelium and a myofibroblastic-like reactive stroma. A...

Cell biology international, Jan 11, 2017

One of the recognized issues in prostate cancer research is the lack of animal models allowing th... more One of the recognized issues in prostate cancer research is the lack of animal models allowing the research of pathological, biochemical, and genetic factors in immunocompetent animals. Our research group has successfully employed the gerbil in several studies for prostate diseases. In the present work, we aimed to analyze the effect of chronic bacterial inflammation on N-methyl-N-nitrosourea (MNU)-induced prostate carcinogenesis in gerbils. Histopathological assessment of the prostatic complex revealed that treatment combinations with MNU plus testosterone or bacterial infection resulted in a promotion of prostate cancer, with bacterial inflammation being more effective in increasing premalignant and malignant tissular alterations than testosterone in the prostate. Furthermore, chronic bacterial inflammation itself induced premalignant lesions in the ventral lobe and increased their frequency in the dorsolateral lobe as well as malignant lesions in the ventral prostate. These anima...

International journal of chronic obstructive pulmonary disease, 2017

In spite of the numerous studies on chronic obstructive pulmonary disease (COPD), the cellular an... more In spite of the numerous studies on chronic obstructive pulmonary disease (COPD), the cellular and molecular basis of the disease's development remain unclear. Neutrophils and eosinophils are known to be key players in COPD. Recently, neutrophil extracellular trap cell death (NETosis), a mechanism due to decondensation and extrusion of chromatin to form extracellular traps, has been demonstrated in COPD. However, there is limited knowledge about eosinophil extracellular trap cell death (EETosis) and its role in the pathogenesis of COPD. The aim of this study was to evaluate EETosis in stable COPD. Induced sputum obtained from healthy smokers and low exacerbation risk COPD A or B group patients or high exacerbation risk COPD C or D group patients were included. Samples were examined using electron microscopy and immunofluorescence. Healthy smokers (n=10) and COPD A (n=19) group exhibited neutrophilic or paucigranulocytic phenotypes, with NETosis being absent in these patients. In...

El presente proyecto de investigacion esta dirigido a obtener informacion integral sobre los meca... more El presente proyecto de investigacion esta dirigido a obtener informacion integral sobre los mecanismos regulatorios involucrados en la secrecion de celulas hipofisiarias. Estos procesos estan modulados por numerosos factores que producen distintos grados de diferenciacion celular, heterogeneidad morfologica y funcional de las diferentes poblaciones, correlacionados con la presencia de variantes moleculares de las hormonas secretadas por cada tipo celular. Para lograr el objetivo propuesto es necesario determinar la variacion de los parametros bioquimicos y ultraestructurales que se produce en respuesta a diferentes agentes estimulatorios o inhibitorios que modifican la secrecion y la poblacion de los distintos tipos celulares de la hipofisis, especificamente sobre las poblaciones de celulas lactotropas y somatotropas. Se desarrollaran los siguientes objetivos especificos: a) Variaciones funcionales de las subpoblaciones de las celulas lactotropas; respuesta diferencial a los neuropeptidos de accion directa TRH AII y de accion paracrina AII y GnRH. b) Regulacion paracrina de prolactina, se estudiara la interaccion de las celulas lactotropas con distintos tipos celulares en cultivos enriquecidos. c) Dinamica de la secrecion de PRL; se estudiara el mecanismo del proceso secretorio de PRL con el uso de drogas inhibitorias de la sintesis o bloqueantes de la secrecion proteica a distintos niveles. d) Factores que participan en la proliferacion y diferenciacion de celulas somatotropas; estudio de los mecanismos por los cuales e se regula la poblacion de celulas somatotropas.

PLOS ONE, 2020

Allergic asthma is the most common phenotype of the pathology, having an early-onset in childhood... more Allergic asthma is the most common phenotype of the pathology, having an early-onset in childhood and producing a Th2-driven airways remodeling process that leads to symptoms and pathophysiological changes. The avoidance of aeroallergen exposure in early life has been shown to prevent asthma, but without repeated success and with the underlying preventive mechanisms at the beginning of asthma far to be fully recognized. In the present study, we aimed to evaluate if neonatal LPS-induced boost in epithelial host defenses contribute to prevent OVA-induced asthma in adult mice. To this, we focused on the response of bronchiolar club cells (CC), which are highly specialized in maintaining the epithelial homeostasis in the lung. In these cells, neonatal LPS administration increased the expression of TLR4 and TNFα, as well as the immunodulatory/antiallergic proteins: club cell secretory protein (CCSP) and surfactant protein D (SP-D). LPS also prevented mucous metaplasia of club cells and reduced the epidermal growth factor receptor (EGFR)-dependent mucin overproduction, with mice displaying normal breathing patterns after OVA challenge. Furthermore, the overexpression of the epithelial Th2-related molecule TSLP was blunted, and normal TSLP and IL-4 levels were found in the bronchoalveolar lavage. A lower eosinophilia was detected in LPS-pretreated mice, along with an increase in phagocytes and regulatory cells (CD4+CD25+FOXP3+ and CD4+IL-10+), together with higher levels of IL-12 and TNFα. In conclusion, our study demonstrates stable asthma-preventive epithelial effects promoted by neonatal LPS stimulation, leading to the presence of regulatory cells in the lung. These anti-allergic dynamic mechanisms would be overlaid in the epithelium, favored by an adequate epidemiological environment, during the development of asthma.

Journal of Cellular Physiology, 2017

Prostatic smooth muscle cells (pSMCs) differentiation is a key factor for prostatic homeostasis, ... more Prostatic smooth muscle cells (pSMCs) differentiation is a key factor for prostatic homeostasis, with androgens exerting multiple effects on these cells. Here, we demonstrated that the myodifferentiator complex Srf/Myocd is up-regulated by testosterone in a dose-dependent manner in primary cultures of rat pSMCs, which was associated to the increase in Acta2, Cnn1, and Lmod1 expressions. Blocking Srf or Myocd by siRNAs inhibited the myodifferentiator effect of testosterone. While LPS led to a dedifferentiated phenotype in pSMCs, characterized by down-regulation of Srf/Myocd and smooth muscle cell (SMC)-restricted genes, endotoxin treatment on Myocd-overexpressing cells did not result in phenotypic alterations. Testosterone at a physiological dose was able to restore the muscular phenotype by normalizing Srf/Myocd expression in inflammation-induced dedifferentiated pSMCs. Moreover, the androgen reestablished the proliferation rate and IL-6 secretion increased by LPS. These results provide novel evidence regarding the myodifferentiating role of testosterone on SMCs by modulating Srf/Myocd. Thus, androgens preserve prostatic SMC phenotype, which is essential to maintain the normal structure and function of the prostate. This article is protected by copyright. All rights reserved.

La Enfermedad Pulmonar Obstructiva Cronica (EPOC) ocupa la sexta posicion entre las causas de mue... more La Enfermedad Pulmonar Obstructiva Cronica (EPOC) ocupa la sexta posicion entre las causas de muerte en todo el mundo, siendo ademas una importante causa de incapacidad, que se ve reflejada en perdidas de la productividad y en altos costos economicos. Por lo tanto, son prioritarias las politicas sanitarias orientadas a frenar el crecimiento y el indice de morbi-mortalidad de la EPOC y de las enfermedades pulmonares en general. La epidemia actual de la EPOC es debida en parte al habito de fumar; sin embargo, solo el 20% de los fumadores la desarrollan, desconociendose, entre otros procesos fisiopatogenicos, las bases celulares y moleculares que determinan que un fumador desarrolle la enfermedad. Este proyecto resulta de la interaccion de profesionales del area clinica, neumonologos del SANATORIO ALLENDE, y del area basica que se desempenan como docentes investigadores en la FACULTAD DE CIENCIAS MEDICAS de la UNC. Nuestro OBJETIVO GENERAL es investigar aspectos aun no evaluados sobre la EPOC, en la busqueda de nuevos parametros adecuados para medir del grado de inflamacion bronquial, que permitan mejorar su prevencion, diagnostico y tratamiento. Una herramienta fundamentalmente usada es la inflamometria del esputo inducido, resultando sencillo, no invasivo y de bajo costo, puesto a punto por primera vez en Argentina por integrantes de esta red. Los objetivos especificos estan orientados a responder diferentes hipotesis planteadas a partir de la problematica diaria del consultorio y desde conocimientos recientes sobre las celulas inflamatorias. El primer aspecto de interes es dilucidar la incidencia de la NETosis, muerte alternativa de los neutrofilos, en el mantenimiento del proceso inflamatorio y de los sintomas propios de la EPOC. Este es un tipo de muerte en el cual se libera el DNA junto al contenido intragranular como mecanismo antimicrobiano y que, segun datos preliminares de nuestro grupo, estaria descontrolado en pacientes con EPOC [18], exponiendo DNA y antigenos propios. De alli nuestros objetivos tendientes en primer lugar a identificar y corroborar NETosis en esputo inducido mediante microscopia confocal y electronica, asociandola con la severidad en la sintomatologia de la EPOC. En segundo lugar se determinara la relacion de NETosis con posibles manifestaciones autoinmunes en la EPOC, para lo cual se determinaran anticuerpos sericos anti-DNA y anticuerpos anti-proteinas del neutrofilo por ELISA. Un tercer objetivo estara orientado mas directamente a analizar la correlacion entre NETosis y el grado de inflamacion de la via aerea, traducido en la relacion entre macrofagos proinflamatorios (M1) y antiinflamatorios (M2) presentes en el esputo inducido, que sera examinada mediante citometria de flujo. Una segunda incognita a abordar proviene de un clasico interrogante en la clinica neumonologica acerca de una posible entidad comun entre la EPOC y el asma. En este contexto, muchos de los pacientes con EPOC tendrian en realidad un fondo asmatico, en el cual el habito tabaquico desencadenaria el proceso inflamatorio. Esta hipotesis se refleja en nuestro ultimo objetivo especifico, investigando marcadores de asma en pacientes con EPOC, a fin de evidenciar un posible solapamiento entre ambas entidades y reorientar la terapia en estos pacientes. En el planteo y ejecucion de estos objetivos, se incluiran comparativamente pacientes con EPOC, fumadores sanos, asmaticos de desarrollo adulto y voluntarios sanos. El desarrollo del presente proyecto impactara significativamente no solo en la identificacion de nuevos blancos terapeuticos que optimicen la resolucion de las enfermedades pulmonares, sino tambien en la formacion de recursos humanos, fortaleciendo vinculos entre los hospitales, los laboratorios de investigacion y las casas de altos estudios de nuestra provincia.

Experimental Biology and Medicine, 2014

Atopic asthma is a chronic allergic disease that involves T-helper type 2 (Th2)-inflammation and ... more Atopic asthma is a chronic allergic disease that involves T-helper type 2 (Th2)-inflammation and airway remodeling. Bronchiolar club cells (CC) and alveolar macrophages (AM) are sentinel cells of airway barrier against inhaled injuries, where allergy induces mucous metaplasia of CC and the alternative activation of AM, which compromise host defense mechanisms and amplify Th2-inflammation. As there is evidence that high levels of environmental endotoxin modulates asthma, the goal of this study was to evaluate if the activation of local host defenses by Lipopolysaccharide (LPS) previous to allergy development can contribute to preserving CC and AM protective phenotypes. Endotoxin stimulus before allergen exposition reduced hallmarks of allergic inflammation including eosinophil influx, Interleukin-4 and airway hyperreactivity, while the T-helper type 1 related cytokines IL-12 and Interferon-γ were enhanced. This response was accompanied by the preservation of the normal CC phenotype a...

Veterinary Ophthalmology, 2009

To describe the functional and structural characteristics of the cornea in healthy Guinea pigs. H... more To describe the functional and structural characteristics of the cornea in healthy Guinea pigs. Healthy male and female pigmented and albino Guinea pigs (Caviaporcellus) aged 3-5 months old were used. The animals&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; corneas underwent different in vivo studies including: slit-lamp biomicroscopy, fluorescein staining (FS), break-up time test (BUT), confocal microscopy and pachymetry. The corneas were also studied histopathologically with light microscopy, immunohistochemistry and transmission electron microscopy. No significant differences were found between pigmented and albino animals, male and female, OD and OS in any study performed. The differences on corneal thickness values were not significant among central (227.85 +/- 14.09 microm) and upper and temporal peripheral regions (226.60 +/- 12.50 and 225.70 +/- 14.40 microm, respectively). All histological studies performed permitted identification and precise description of the different corneal structures in Guinea pigs: the stratified epithelium (45.52 +/- 5.26 microm), Bowman&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s layer (2.23 +/- 0.38 microm), stroma (163.69 +/- 4.90 microm), Descemet&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s membrane (3.96 +/- 0.46 microm) and the endothelium (5.09 +/- 0.71 microm). Combining results from all eyes mean and SD from corneal BUT values was 4.98 +/- 1.67 s. Corneas often showed discrete superficial erosions being the FS positive in both eyes from all the animals. This study provides a detailed in vivo and postfixed histological description of the Guinea pig&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s cornea and information about the physiological tests.

International Journal of Experimental Pathology, 2013

Histochemistry and Cell Biology, 2008

The objectives of the present work were to assess whether epithelial cells from the diVerent segm... more The objectives of the present work were to assess whether epithelial cells from the diVerent segments of epididymis express TR 1-1 isoforms, to depict its subcellular immunolocalization and to evaluate changes in their expression in rats experimentally submitted to a hypothyroid state by injection of 131I. In euthyroid and hypothyroid groups, TR protein was expressed in epididymal epithelial cells, mainly in the cytoplasmic compartment while only a few one showed a staining in the nucleus as well. A similar TR immunostaining pattern was detected in the diVerent segments of the epididymis. In hypothyroid rats, the number of TR-immunoreactive epithelial cells as well as the intensity of the cytoplasmic staining signiWcantly increased in all sections analyzed. In consonance to the immunocytochemical analysis, the expression of TR 1-1 isoforms, assessed by Western blot revealed signiWcantly higher levels of TR in cytosol compared to the nuclear fractions. Furthermore, TR expression of both 1 and 1 isoforms and their mRNA levels were increased by the hypothyroid state. The immuno-electron-microscopy showed speciWc reaction for TR in principal cells associated with eucromatin, cytosolic matrix and mitochondria. The diVerences in expression levels assessed in control and thyroidectomized rats ascertain a speciWc function of TH on this organ.

… and Cell Biology, 2008

Histochem Cell Biol (2008) 129:631–642 DOI 10.1007/s00418-008-0397-8 ... Thyroid hormone receptor... more Histochem Cell Biol (2008) 129:631–642 DOI 10.1007/s00418-008-0397-8 ... Thyroid hormone receptor 1 – 1 expression in epididymal ... Ana Lucía De Paul · Jorge Humberto Mukdsi · Claudia Gabriela Pellizas · María Montesinos · Silvina Gutiérrez · Sebastián Susperreguy · Alberto ...

Applications of Immunocytochemistry, 2012

Applications of Immunocytochemistry 66 appropriate selection of an embedding resin and the ready ... more Applications of Immunocytochemistry 66 appropriate selection of an embedding resin and the ready availability of the specific antibodies for the molecules whose ultrastructural location needs to be determined.

A pesar de los avances fisiopatogenicos y terapeuticos, el asma permanece sin control. El objetiv... more A pesar de los avances fisiopatogenicos y terapeuticos, el asma permanece sin control. El objetivo de este ensayo es mostrar evidencias clinicas-cientificas que los instrumentos utilizados en el manejo del asma no son herramientas suficientes y proponer una estrategia para el abordaje y su control en forma objetiva, teniendo como blanco terapeutico la inflamacion de la via aerea segun parametros celulares del esputo inducido. Evaluamos en 15 pacientes asmaticos bajo tratamiento: el grado de control del asma (GCA) en base a criterios clinicos, espirometricos y el cuestionario de control, segun recomendaciones de GINA (Global Initiative for Asthma). Posteriormente todos los pacientes fueron sometidos a la recoleccion de una muestra de esputo inducido (EI) para realizar el recuento diferencial de celulas segun tecnica previamente validada. Del total estudiados, obtuvimos que 7 pacientes, a pesar que se encontraban totalmente controlados segun parametros de GCA 5 presentaban un perfil i...

Revista de la Facultad de Ciencias Médicas de Córdoba

Objetivo: Evaluar la prevalencia de riesgo de asma en adolescentes expuestos a ceniza volcánica a... more Objetivo: Evaluar la prevalencia de riesgo de asma en adolescentes expuestos a ceniza volcánica a un año de la erupción del Volcán Calbuco, en 2 ciudades con diferentes grado de afectación. Métodologia: Estudio de Corte Transversal, en adolescentes de 13-14 años en dos ciudades con diferente grado de exposición a ceniza volcánica: San Carlos de Bariloche y Cipolletti. Se aplicó cuestionario y video-cuestionario ISAAC para determinar riesgo de asma. Las variables demográficas y percepción de exposición, se obtuvieron por cuestionario previo a la aplicación de metodología ISAAC. Resultados: Se encuestaron 511 sujetos en total. Se reportó prevalencia de síntomas de riesgo de asma en Bariloche resultó de 14% comparado con Cipolletti que reportó 10% (p=0.32). Se informó mayor exposición a ceniza volcánica en la ciudad de Bariloche vs. Cipolletti; 14% y 6% respectivamente con p<0.05. Conclusiones:La falta diferencia estadisticamente significativa respecto a la prevalencia de sintomas d...

Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology

Glutamine (GLN) avoids the inhibition of the intestinal Ca2+ absorption caused by menadione (MEN)... more Glutamine (GLN) avoids the inhibition of the intestinal Ca2+ absorption caused by menadione (MEN) through oxidative stress. The purpose of this study was to elucidate whether molecules of transcellular and/or paracellular pathways of intestinal Ca2+ absorption are involved in the GLN action and underlying mechanisms. One-month old chicks were divided in four groups: 1) controls, 2) MEN treated, 3) GLN treated and 4) GLN + MEN treated. The morphology of intestinal villi, the intestinal Ca2+ absorption and the molecules involved in the transcellular and paracellular pathways were analyzed. Markers of autophagy and inflammation were also evaluated. The data demonstrated that GLN protected both transcellular and paracellular pathways. GLN avoided morphological changes in the intestine caused by MEN. GLN protected the gene expression of transporters involved in the transcellular pathway and the gene and protein expression of molecules belonging to the paracellular pathways altered by MEN. GLN increased the LC3-II protein expression and the number of acidic vesicular organelles, markers of autophagy, and blocked an increase in the NFkB protein expression in the nuclei and in the IL-6 gene expression caused by MEN. In conclusion, GLN protects both transcellular and paracellular pathways of intestinal Ca2+ absorption by increasing autophagy and blocking inflammation.

Frontiers in immunology, 2018

Neutrophils are major effectors of acute inflammation against infection and tissue damage, with a... more Neutrophils are major effectors of acute inflammation against infection and tissue damage, with ability to adapt their phenotype according to the microenvironment. Although sex hormones regulate adaptive immune cells, which explains sex differences in immunity and infection, little information is available about the effects of androgens on neutrophils. We therefore aimed to examine neutrophil recruitment and plasticity in androgen-dependent and -independent sites under androgen manipulation. By using a bacterial model of prostate inflammation, we showed that neutrophil recruitment was higher in testosterone-treated rats, with neutrophil accumulation being positively correlated to serum levels of testosterone and associated to stronger inflammatory signs and tissue damage. Testosterone also promoted LPS-induced neutrophil recruitment to the prostate, peritoneum, and liver sinusoids, as revealed by histopathology, flow cytometry, and intravital microscopy. Strikingly, neutrophils in p...

Frontiers in immunology, 2018

Galectins, a family of animal lectins characterized by their affinity for N-acetyllactosamine-enr... more Galectins, a family of animal lectins characterized by their affinity for N-acetyllactosamine-enriched glycoconjugates, modulate several immune cell processes shaping the course of innate and adaptive immune responses. Through interaction with a wide range of glycosylated receptors bearing complex branched N-glycans and core 2-O-glycans, these endogenous lectins trigger distinct signaling programs thereby controling immune cell activation, differentiation, recruitment and survival. Given the unique features of mucosal inflammation and the differential expression of galectins throughout the gastrointestinal tract, we discuss here key findings on the role of galectins in intestinal inflammation, particularly Crohn's disease, ulcerative colitis, and celiac disease (CeD) patients, as well as in murine models resembling these inflammatory conditions. In addition, we present new data highlighting the regulated expression of galectin-1 (Gal-1), a proto-type member of the galectin famil...

Molecular cancer research : MCR, Jan 9, 2018

Emerging evidence suggests that unregulated Toll-like receptors (TLRs) signaling promotes tumor s... more Emerging evidence suggests that unregulated Toll-like receptors (TLRs) signaling promotes tumor survival signals, thus favoring tumor progression. Here, the mechanism underlying TLR4 overexpression in papillary thyroid carcinomas (PTCs) mainly harboring the BRAFV600E mutation was studied. TLR4 was overexpressed in PTCs compared to non-neoplastic thyroid tissue. Moreover, paired clinical specimens of primary PTC and its lymph node metastasis showed a significant upregulation of TLR4 levels in the metastatic tissues. In agreement, conditional BRAFV600E expression in normal rat thyroid cells and mouse thyroid tissue upregulated TLR4 expression levels. Furthermore, functional TLR4 expression was demonstrated in PTC cells by increased NF-κB transcriptional activity in response to the exogenous TLR4-agonist lipopolysaccharide (LPS). Of note, The Cancer Genome Atlas (TCGA) data analysis revealed that BRAFV600E-positive tumors with high TLR4 expression were associated with shorter disease-f...

Cell biology international, Jan 28, 2017

The prostate gland is a strictly androgen-dependent organ which is also the main target of infect... more The prostate gland is a strictly androgen-dependent organ which is also the main target of infectious and inflammatory diseases in the male reproductive tract. Host defenses and immunity of the gland have unique features to maintain a constant balance between response and tolerance to diverse antigens. In this context, the effects of reproductive hormones on the male tract are thus complex and have just started to be defined. From the classical description of "the prostatic antibacterial factor," many host defense proteins with potent microbicidal and anti-tumoral activities have been described in the organ. Indeed, it has been proposed a central role for resident cells, that is, epithelial and smooth muscle cells, in the prostatic response against injuries. However, these cells also represent the target of the inflammatory damage, leading to the development of a Proliferative Inflammatory Atrophy-like process in the epithelium and a myofibroblastic-like reactive stroma. A...

Cell biology international, Jan 11, 2017

One of the recognized issues in prostate cancer research is the lack of animal models allowing th... more One of the recognized issues in prostate cancer research is the lack of animal models allowing the research of pathological, biochemical, and genetic factors in immunocompetent animals. Our research group has successfully employed the gerbil in several studies for prostate diseases. In the present work, we aimed to analyze the effect of chronic bacterial inflammation on N-methyl-N-nitrosourea (MNU)-induced prostate carcinogenesis in gerbils. Histopathological assessment of the prostatic complex revealed that treatment combinations with MNU plus testosterone or bacterial infection resulted in a promotion of prostate cancer, with bacterial inflammation being more effective in increasing premalignant and malignant tissular alterations than testosterone in the prostate. Furthermore, chronic bacterial inflammation itself induced premalignant lesions in the ventral lobe and increased their frequency in the dorsolateral lobe as well as malignant lesions in the ventral prostate. These anima...

International journal of chronic obstructive pulmonary disease, 2017

In spite of the numerous studies on chronic obstructive pulmonary disease (COPD), the cellular an... more In spite of the numerous studies on chronic obstructive pulmonary disease (COPD), the cellular and molecular basis of the disease's development remain unclear. Neutrophils and eosinophils are known to be key players in COPD. Recently, neutrophil extracellular trap cell death (NETosis), a mechanism due to decondensation and extrusion of chromatin to form extracellular traps, has been demonstrated in COPD. However, there is limited knowledge about eosinophil extracellular trap cell death (EETosis) and its role in the pathogenesis of COPD. The aim of this study was to evaluate EETosis in stable COPD. Induced sputum obtained from healthy smokers and low exacerbation risk COPD A or B group patients or high exacerbation risk COPD C or D group patients were included. Samples were examined using electron microscopy and immunofluorescence. Healthy smokers (n=10) and COPD A (n=19) group exhibited neutrophilic or paucigranulocytic phenotypes, with NETosis being absent in these patients. In...

El presente proyecto de investigacion esta dirigido a obtener informacion integral sobre los meca... more El presente proyecto de investigacion esta dirigido a obtener informacion integral sobre los mecanismos regulatorios involucrados en la secrecion de celulas hipofisiarias. Estos procesos estan modulados por numerosos factores que producen distintos grados de diferenciacion celular, heterogeneidad morfologica y funcional de las diferentes poblaciones, correlacionados con la presencia de variantes moleculares de las hormonas secretadas por cada tipo celular. Para lograr el objetivo propuesto es necesario determinar la variacion de los parametros bioquimicos y ultraestructurales que se produce en respuesta a diferentes agentes estimulatorios o inhibitorios que modifican la secrecion y la poblacion de los distintos tipos celulares de la hipofisis, especificamente sobre las poblaciones de celulas lactotropas y somatotropas. Se desarrollaran los siguientes objetivos especificos: a) Variaciones funcionales de las subpoblaciones de las celulas lactotropas; respuesta diferencial a los neuropeptidos de accion directa TRH AII y de accion paracrina AII y GnRH. b) Regulacion paracrina de prolactina, se estudiara la interaccion de las celulas lactotropas con distintos tipos celulares en cultivos enriquecidos. c) Dinamica de la secrecion de PRL; se estudiara el mecanismo del proceso secretorio de PRL con el uso de drogas inhibitorias de la sintesis o bloqueantes de la secrecion proteica a distintos niveles. d) Factores que participan en la proliferacion y diferenciacion de celulas somatotropas; estudio de los mecanismos por los cuales e se regula la poblacion de celulas somatotropas.