783 resultados para cordón umbilical
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Down syndrome (DS) is a common birth defect characterized by the trisomy of chromosome 21. DS-affected umbilical cords (UCs) of fetuses show altered architecture of the extracellular matrix. Overexpression of the chromosome 21 genes encoding the collagen type VI (COLVI) chains α1(VI) and α2(VI), COL6A1 and COL6A2, respectively, has also reported to occur in the nuchal skin of DS fetuses. The aim of this study was therefore to evaluate the COLVI content in euploid and DS-affected UCs and human skin fibroblasts, and to investigate the relationships between COLVI and hyaluronan (HA) and HA synthase-2 (HAS2). We found that the UCs of DS fetuses showed denser staining of COLVI and increased COL6A2 expression at both early and term gestational ages. In vitro expression studies in DS-derived fibroblasts showed similarly increased amounts of α1(VI) and α2(VI) chains at the protein and transcriptional level, supporting the hypothesis of the gene dosage effect. Furthermore, increased levels of HA and HAS2 were also found in DS-derived skin fibroblast cultures. Notably, silencing of COL6A2 in DS-derived cells resulted in downregulation of HAS2, with a simultaneous decrease in secreted HA. Exogenous addition of COLVI to normal fibroblasts did not have any effect on HAS2 expression. In conclusion, UCs and skin fibroblasts in DS show significant increases in COLVI and HA; the overexpression of COL6A2 in DS tissue and cells is closely related to the increased expression of HAS2. These data may explain the DS phenotypes and their effects in organ tissue maturation.
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INTRODUCTION 17β-estradiol (E2) has been found to induce vasodilation in the cardiovascular system and at physiological levels, resulting in prevention of cerebral vasospasm following subarachnoid hemorrhage (SAH) in animal models. The goal of this study was to analyze the cellular mechanism of nitric oxide (NO) production and its relation to E2, in vitro in brain and peripheral endothelial cells. METHODS Human umbilical endothelial cells (HUVEC) and brain endothelial cells (bEnd.3) were treated with estradiol (E2, 0.1, 10, 100, and 1,000 nM), and supernatant was collected at 0, 5, 15, 30, 60, and 120 min for nitric oxide metabolome (nitrite, NO₂) measurements. Cells were also treated with E2 in the presence of 1400W, a potent eNOS inhibitor, and ICI, an antagonist of estradiol receptors (ERs). Effects of E2 on eNOS protein expression were assessed with Western blot analysis. RESULTS E2 significantly increased NO2 levels irrespective of its concentration in both cell lines by 35 % and 42 % (p < 0.05). The addition of an E2 antagonist, ICI (10 μM), prevented the E2-induced increases in NO2 levels (11 % p > 0.05). The combination of E2 (10 nM) and a NOS inhibitor (1400W, 5 μM) inhibited NO2 increases in addition (4 %, p > 0.05). E2 induced increases in eNOS protein levels and phosphorylated eNOS (eNOS(p)). CONCLUSIONS This study indicates that E2 induces NO level increases in cerebral and peripheral endothelial cells in vitro via eNOS activation and through E2 receptor-mediated mechanisms. Further in vivo studies are warranted to evaluate the therapeutic value of estrogen for the treatment of SAH-induced vasospasm.
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Fetal serum cholesterol and lipoprotein concentrations differ between preterm and term born neonates. An imbalance of the flow of cholesterol from the sites of synthesis or efflux from cells of peripheral organs to the liver, the reverse cholesterol transport (RCT), is linked to atherosclerosis and cardiovascular disease (CVD). Preterm delivery is a risk factor for the development of CVD. Thus, we hypothesized that RCT is affected by a diminished cholesterol acceptor capacity in preterm as compared to term fetuses. Cholesterol efflux assays were performed in RAW264.7, HepG2, and HUVEC cell lines. In the presence and absence of ABC transporter overexpression by TO-901317, umbilical cord sera of preterm and term born neonates (n = 28 in both groups) were added. Lipid components including high density lipoprotein (HDL), low density lipoprotein (LDL), apolipoprotein A1, and apolipoprotein E were measured and related to fractional cholesterol efflux values. We found overall, fractional cholesterol efflux to remain constant between the study groups, and over gestational ages at delivery, respectively. However, correlation analysis revealed cholesterol efflux values to be predominantly related to HDL concentration at term, while in preterm neonates, cholesterol efflux was mainly associated with LDL In conclusion cholesterol acceptor capacity during fetal development is kept in a steady state with different mechanisms and lipid fractions involved at distinct stages during the second half of fetal development. However, RCT mechanisms in preterm neonates seem not to be involved in the development of CVD later in life suggesting rather changes in the lipoprotein pattern causative.
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The discovery of mesenchymal stem cells (MSCs) in perinatal sources, such as the amniotic fluid (AF) and the umbilical connective tissue, the so-called Wharton's jelly (WJ), has transformed them into promising stem cell grafts for the application in regenerative medicine. The advantages of AF-MSCs and WJ-MSCs over adult MSCs, such as bone marrow-derived mesenchymal stem cells (BMMSCs), include their minimally invasive isolation procedure, their more primitive cell character without being tumourigenic, their low immunogenicity and their potential autologous application in congenital disorders and when cryopreserved in adulthood. This chapter gives an overview of the biology of AF-MSCs and WJMSCs, and their regenerative potential based on the results of recent preclinical and clinical studies. In the end, open questions concerning the use of WJ-MSCs and AF-MSCs in regenerative medicine will be emphasized.
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BACKGROUND Platelet-rich concentrates are used as a source of growth factors to improve the healing process. The diverse preparation protocols and the gaps in knowledge of their biological properties complicate the interpretation of clinical results. QUESTIONS/PURPOSES In this study we aimed to (1) analyze the concentration and kinetics of growth factors released from leukocyte- and platelet-rich fibrin (L-PRF), leukocyte- and platelet-rich plasma (L-PRP), and natural blood clot during in vitro culture; (2) investigate the migration of mesenchymal stem cells (MSCs) and human umbilical vein endothelial cells (HUVECs) as a functional response to the factors released; and (3) uncover correlations between individual growth factors with the initial platelet/leukocyte counts or the induced cell migration. METHODS L-PRF, L-PRP, and natural blood clot prepared from 11 donors were cultured in vitro for 28 days and media supernatants collected after 8 hours and 1, 3, 7, 14, and 28 days. Released transforming growth factor β1 (TGF-β1), vascular endothelial growth factor (VEGF), insulin growth factor (IGF-1), platelet-derived growth factor AB (PDGF-AB), and interleukin-1β (IL-1β) were measured in the supernatants with enzyme-linked immunosorbent assay. Migration of MSC and HUVEC induced by the supernatants was evaluated in Boyden chambers. RESULTS More TGF-ß1 was released (mean ± SD in pg/mL of blood) from L-PRF (37,796 ± 5492) compared with L-PRP (23,738 ± 6848; p < 0.001) and blood clot (3739 ± 4690; p < 0.001), whereas more VEGF and IL-1ß were released from blood clot (1933 ± 704 and 2053 ± 908, respectively) compared with both L-PRP (642 ± 208; p < 0.001 and 273 ± 386; p < 0.001, respectively) and L-PRF (852 ± 376; p < 0.001 and 65 ± 56, p < 0.001, respectively). No differences were observed in IGF-1 and PDGF-AB released from any of the concentrates. TGF-β1 release peaked at Day 7 in L-PRF and at 8 hours and Day 7 in L-PRP and 8 hours and Day 14 in blood clot. In all concentrates, main release of VEGF occurred between 3 and 7 days and of IL-1β between Days 1 and 7. IGF-1 and PDGF-AB were released until Day 1 in L-PRP and blood clot, in contrast to sustained release over the first 3 days in L-PRF. The strongest migration of MSC occurred in response to L-PRF, and more HUVEC migration was seen in L-PRF and blood clot compared with L-PRP. TGF-β1 correlated with initial platelet counts in L-PRF (Pearson r = 0.66, p = 0.0273) and initial leukocyte counts in L-PRP (Pearson r = 0.83, p = 0.0016). A positive correlation of IL-1β on migration of MSC and HUVEC was revealed (Pearson r = 0.16, p = 0.0208; Pearson r = 0.31, p < 0.001). CONCLUSIONS In comparison to L-PRP, L-PRF had higher amounts of released TGF-β1, a long-term release of growth factors, and stronger induction of cell migration. Future preclinical studies should confirm these data in a defined injury model. CLINICAL RELEVANCE By characterizing the biologic properties of different platelet concentrates in vitro, we may gain a better understanding of their clinical effects and develop guidelines for specific future applications.
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In vitro engineered tissues which recapitulate functional and morphological properties of bone marrow and bone tissue will be desirable to study bone regeneration under fully controlled conditions. Among the key players in the initial phase of bone regeneration are mesenchymal stem cells (MSCs) and endothelial cells (ECs) that are in close contact in many tissues. Additionally, the generation of tissue constructs for in vivo transplantations has included the use of ECs since insufficient vascularization is one of the bottlenecks in (bone) tissue engineering. Here, 3D cocultures of human bone marrow derived MSCs (hBM-MSCs) and human umbilical vein endothelial cells (HUVECs) in synthetic biomimetic poly(ethylene glycol) (PEG)-based matrices are directed toward vascularized bone mimicking tissue constructs. In this environment, bone morphogenetic protein-2 (BMP-2) or fibroblast growth factor-2 (FGF-2) promotes the formation of vascular networks. However, while osteogenic differentiation is achieved with BMP-2, the treatment with FGF-2 suppressed osteogenic differentiation. Thus, this study shows that cocultures of hBM-MSCs and HUVECs in biological inert PEG matrices can be directed toward bone and bone marrow-like 3D tissue constructs.
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BACKGROUND Pancreatic stone protein (PSP) has been identified as a promising sepsis marker in adults, children and neonates. However, data on population-based reference values are lacking. This study aimed to establish age-specific reference values for PSP. METHODS PSP was determined using a specific ELISA. PSP serum concentrations were determined in 372 healthy subjects including 217 neonates, 94 infants and children up to 16 years, and 61 adults. The adjacent categories method was used to determine which age categories had significantly different PSP concentrations. RESULTS PSP circulating levels were not gender-dependent and ranged from 1.0 to 99.4 ng/ml with a median of 9.2 ng/ml. PSP increased significantly between the age categories, from a median of 2.6 ng/ml in very preterm newborns, to 6.3 ng/ml in term newborns, to 16.1 ng/ml in older children (p < 0.001). PSP levels were higher on postnatal day three compared to levels measured immediately post delivery (p < 0.001). Paired umbilical artery and umbilical vein samples were strongly correlated (p < 0.001). Simultaneously obtained capillary heel-prick versus venous samples showed a good level of agreement for PSP (Rho 0.89, bias 19 %). CONCLUSIONS This study provides age-specific normal values that may be used to define cut-offs for future trials on PSP. We demonstrate an age-dependent increase of PSP from birth to childhood.
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Upon activation, platelets release plasma-membrane derived microparticles (PMPs) exposing phosphatidylserine (PS) on their surface. The function and clearance mechanism of these MPs are incompletely understood. As they are pro-coagulant and potentially pro-inflammatory, rapid clearance from the circulation is essential for prevention of thrombotic diseases. The tyrosine kinase receptors Tyro3, Axl and Mer (TAMs) and their ligands protein S and Gas6 are involved in the uptake of PS-exposing apoptotic cells in macrophages and dendritic cells. Both TAMs and their ligands are expressed in the vasculature, the functional significance of which is poorly understood. In this study we investigated how vascular TAMs and their ligands may mediate endothelial uptake of PMPs. PMPs, generated from purified human platelets, were isolated by ultracentrifugation and labeled with biotin or PKH67. The uptake of labeled MPs in the presence of protein S and Gas6 in human aortic endothelial cells (HAEC) and human umbilical vein endothelial cells (HUVEC) was monitored by flow cytometry, western blotting and confocal/electron microscopy. We found that both endothelial cell types can phagocytose PMPs, and using TAM-blocking antibodies or siRNA knock-down of individual TAMs we show that the uptake is mediated by endothelial Axl and Gas6. As circulating PMPs-levels were not altered in Gas6-/- mice compared to Gas6+/+ mice, we hypothesize that the Gas6-mediated uptake is not a means to clear the bulk of circulating PMPs but may serve to phagocytose PMPs locally generated at sites of platelet activation and as a way to affect endothelial responses.
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Nuclear imaging is used for non-invasive detection, staging and therapeutic monitoring of tumors through the use of radiolabeled probes. Generally, these probes are used for applications in which they provide passive, non-specific information about the target. Therefore, there is a significant need for actively-targeted radioactive probes to provide functional information about the site of interest. This study examined endostatin, an endogenous inhibitor of tumor angiogenesis, which has affinity for tumor vasculature. The major objective of this study was to develop radiolabeled analogues of endostatin through novel chemical and radiochemical syntheses, and to determine their usefulness for tumor imaging using in vitro and in vivo models of vascular, mammary and prostate tumor cells. I hypothesize that this binding will allow for a non-invasive approach to detection of tumor angiogenesis, and such detection can be used for therapeutic monitoring to determine the efficacy of anti-angiogenic therapy. ^ The data showed that endostatin could be successfully conjugated to the bifunctional chelator ethylenedicysteine (EC), and radiolabeled with technetium-99m and gallium-68, providing a unique opportunity to use a single precursor for both nuclear imaging modalities: 99mTc for single photon emission computed tomography and 68Ga for positron emission tomography, respectively. Both radiolabeled analogues showed increased binding as a function of time in human umbilical vein endothelial cells and mammary and prostate tumor cells. Binding could be blocked in a dose-dependent manner by unlabeled endostatin implying the presence of endostatin receptors on both vascular and tumor cells. Animal biodistribution studies demonstrated that both analogues were stable in vivo, showed typical reticuloendothelial and renal excretion and produced favorable absorbed organ doses for application in humans. The imaging data provide evidence that the compounds quantitate tumor volumes with clinically-useful tumor-to-nontumor ratios, and can be used for treatment follow-up to depict changes occurring at the vascular and cellular levels. ^ Two novel endostatin analogues were developed and demonstrated interaction with vascular and tumor cells. Both can be incorporated into existing nuclear imaging platforms allowing for potential wide-spread clinical benefit as well as serving as a diagnostic tool for elucidation of the mechanism of action of endostatin. ^
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Three studies examined seasonal or circadian variations in selected responses to influenza infection or vaccination. The first, a seroepidemiologic study, evaluated temporal patterns of antibody titers to influenza A/Texas. Human umbilical cord bloods were sampled over a two-year period when the virus was not present in the community. No endogenous seasonal pattern was detected. The second study included three experiments on circadian rhythms in mice. Neither susceptibility nor protection from inactivated or attenuated vaccine varied significantly according to time of administration. A slight effect, however, was suggested with inactivated vaccine. Three human vaccine trials comprised the third study. Outcome variables included rise in antibody titer, final antibody titer, incidence of adverse reactions, and protection from community infection. Patterns in antibody response and protection variables were inconsistent, and generally not clinically significant. Local reactions to inactivated vaccine were more frequent if injections were received in the afternoon as compared to morning. This was true to adults that had been previously vaccinated. ^
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En el primer punto de este informe final se realizó un estado del arte y un análisis monográfico del problema que abordamos al que denominamos La Ciudad como Objeto de Estudio, es decir un marco conceptual que nos permita situar el problema abordado además de señalar los avances en cuanto a la gestión de la ciudad. Al tratarse de un análisis teórico abordamos largamente diversos aspectos que se vinculan a la construcción epistemológica del tema. Algunos de los temas tratados son: el futuro de las ciudades en el proceso globalizador, la relación entre campo-ciudad, ciudad y estado nacional, Nación y fronteras nacionales, las teorías del desarrollo y la exclusión social, entre otras cuestiones. En el segundo punto tratamos y profundizamos la descripción de el Proyecto Yacyretá y las Obras Civiles en la Ciudad teniendo en cuenta que este proyecto modifico sustancialmente la urbanización de Posadas, la gestión municipal y la estructura social local. En este sentido se hace referencia a los proyectos del ente público que directamente se desarrollan en el área urbana, denominado en el proyecto como Obras Complementarias, se describe y evalúa ahí las diferentes proyecciones elaboradas por los diferentes momentos de los diferentes gerenciamientos que se sucedieron en el tiempo, a la vez se trata de explicitar las condiciones históricas en que se proyectaron y realizaron las obras. En el tercer punto "El entorno regional y los involucrados" se describe el entorno social del emplazamiento de las obras de análisis a la vez de los actores directa e indirectamente involucrados. Entre otras cuestiones se analizan los indicadores económicos, las características y cambios de la pobreza urbana y rural, el mercado inmobiliario, la infraestructura y los servicios, los sistemas de transporte: terrestre, fluvial y ferroviario, Actores Sociales, entre otros temas. En el punto cuarto se pone el foco de trabajo en la Ciudad de Posadas. Se vincula esta con la dinámica de frontera con su "gemela" ciudad de Encarnación, República del Paraguay que también es modificada por el mismo proyecto y en análogas dimensiones, tanto por su vinculación histórico-geográfico como por sus transformaciones a la luz de las obras del Proyecto Yacyretá. Se realiza una historización de la división y uso del suelo en Posadas, el proyecto Yacyretá y sus efectos en la dinamización del mercado inmobiliario, las políticas municipales y el problema de la vivienda. El punto quinto hace referencia a cual es la percepción de los actores y ciudadanos de Posadas con respecto a la ciudad, la gestión local, lo servicios públicos, etc. Se han adoptado diferentes técnicas en un diseño técnico metodológico que permita triangular técnicas cualitativas y cuantitativas. En tal sentido se ha trabajado diferentes técnicas que a la vez nos permita muestrear y elaborar tipos actores y condiciones de la población en relación al asentamiento en el que se sitúan. Primero exponemos los resultados de una encuesta realizada en la ciudad de Posadas sobre la visión de los ciudadanos respecto a los problemas de la ciudad y las responsabilidades institucionales. Segundo se hace referencia a información cualitativa recogida en lo que denominamos el primer cordón o conglomerado urbano, conformado por una heterogeneidad de barrios de NES medio y bajo. Las técnicas utilizadas en este punto fueron tres grupos focalizados. Tercero, se trata del procesamiento de una encuesta realizada en barrios de construcción pública en Posadas y Garupá, ambos conforman el gran conglomerado metropolitano de Posadas aunque se trate de dos municipios diferentes. Cuarto se analizar allí talleres realizados con población vulnerable de barrios de construcción pública. En las conclusiones se realiza una interpretación de los datos construidos. Durante todo el proceso de investigación el equipo de trabajo colaboró en todos los momentos y fases de investigación, desde el diseño y elaboración del proyecto, la construcción del objeto (problemas, estado del arte y elaboración de hipótesis), el diseño técnico metodológico en el que se fijaron a las estrategias o combinación de técnicas de recolección de información, (información secundaria, encuestas, entrevistas, grupos focales y talleres) procesamiento e interpretación. En las conclusiones se elaboran interpretaciones mediante técnicas del planeamiento estratégico como matriz DAFO, análisis de Juego de Actores, Árbol de Problemas y análisis prospectivo. Pero fundamentalmente se realiza una evaluación territorial de clases sociales. Se incluyen ilustraciones fotográficas y mapas temáticos en algunos puntos del informe.
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Hasta el momento se ha recogido información secundaria, se ha referenciado, tomado nota, escaneado y/o fotocopiado a fin de procesar en el siguiente periodo aunque se ha avanzado ya en ese sentido. Al ser información cualitativa su procesamiento es del mismo modo. La información recogida se procesa también con planos en formato GIS (Sistema de Información Geográfico con programa ARCGIS).La información geoprocesada y editada en planos temáticos se entregaran con el informe final. La información recogida y en principio procesada hace referencia al análisis de acciones y proyectos de inversión pública implementada y por implementarse en Posadas y áreas metropolitanas y al balance de pérdidas-reposiciones desde los aspectos económico-físico-territorial. Hasta el presente se han desarrollado los siguientes aspectos:Obras civiles Yayreta en Posadas y su influencia en el desarrollo de la ciudad; Encuesta realizada en la ciudad de Posadas sobre la visión de los ciudadanos respecto a los problemas de la ciudad y las responsabilidades institucionales; Información cualitativa recogida en lo que denominamos el primer cordón o conglomerado urbano, conformado por una heterogeneidad de barrios de NES medio y bajo. Las técnicas utilizadas en este punto fueron tres grupos focalizados; Se trata del procesamiento de una encuesta realizada en barrios de construcción pública en Posadas y Garupa, ambos conforman el gran conglomerado metropolitano de Posadas aunque se trate de dos municipios diferentes; Se analiza allí talleres realizados con población vulnerable de barrios de construcción pública; Se realiza un ensayo que permita una interpretación conceptual del problema abordado.
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En Mendoza es cada vez más común ralear racimos con el propósito de afectar la composición de las uvas. No obstante, el conocimiento local sobre cómo lograr un equilibrio adecuado de los distintos atributos de calidad mediante el raleo es escaso. El objetivo de esta investigación fue evaluar la relación entre el raleo de racimos en diferentes intensidades y épocas, y los componentes del rendimiento y la composición fenólica de la uva. Para este estudio, que se realizó en un viñedo de Agrelo, Luján de Cuyo, Mendoza (Argentina), se eligió la cultivar Malbec por ser el cepaje emblemático de Argentina y el típico de la Denominación de Origen Controlado (DOC) Luján de Cuyo. En las plantas de dicha cultivar, conducidas en espaldero alto y podadas en cordón Royat bilateral, el raleo fue manual, en tres momentos del ciclo: 1) cuando los granos tenían el tamaño de una arveja; 2) en envero y 3) a 21 °Brix. La intensidad de raleo fue de 25 y 50 % de los racimos. Se comprobó la hipótesis planteada en relación con que el raleo aumenta el tamaño de la baya y mejora la calidad de la uva, por cuanto incrementa la biosíntesis de los polifenoles. En los componentes del rendimiento aumenta el peso del racimo y el tamaño de la baya cuando el raleo se hace temprano y con una intensidad elevada. En cuanto a la influencia en la biosíntesis de los polifenoles se demuestra que el raleo temprano e intenso mejora la concentración de antocianas, catequinas y proantocianidinas. La concentración azucarina se vio incrementada cuando el raleo se hizo en envero y fue intenso.
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Para determinar la influencia del manejo del suelo sobre el microclima de la canopia se estableció un experimento en un viñedo de Cabernet Sauvignon conducido en doble cordón de pitones en espladero alto. Se aplicaron tres tratamientos: Testigo (TR)-suelo sin maleza; (CVP)-cobertura de suelo espontánea y control del desarrollo vegetativo por desbrozado y (CA)-cobertura de flora espontánea y control del desarrollo vegetativo con aplicación de herbicida de contacto desecante. Se midió la radiación fotosintéticamente activa (PAR), la composición espectral de la radiación directa y reflejada, y la temperatura a nivel de racimos en el interior de la canopia. El TR reflejó más radiación en la zona del UV y del visible. El CVP reflejó entre un 8 y 9%, el CA entre el 16 y 18% y el TR entre 18 y 19%. En la radiación reflejada, la relación rojo/rojo lejano fue mayor en CA y TR, mientras que la relación azul/rojo fue mayor en CVP. La temperatura en el interior de la canopia fue mayor en TR (30,8°C) que en CVP (26,7°C). La utilización de coberturas vegetales de raíces superficiales y permanentemente desbrozadas sería de utilidad para reducir la radiación reflejada y la temperatura de la canopia sin competir con el viñedo por agua y nutrimentos.
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Regiones y Departamentos. Relatos de nuestra identidad es una serie que aborda las características turísticas, culturales e históricas de la provincia de Mendoza. En los 14 programas se han destacado y revalorizado las particularidades de cada uno de los departamentos de esta provincia argentina que se dividen en distintas regiones: Valle de Uco, Gran Mendoza, Zona Sur, Zona Este y Zona Nordeste VALLE DE UCO II es un documental que nos invita a seguir conociendo las características más relevantes de los departamentos de Tunuyán, Tupungato y San Carlos. En el bloque 1 repasaremos la historia del departamento de Tunuyán, zona que durante la época colonial se caracterizaba por su desolación. Más tarde y con la llegada del ferrocarril entre otras cosas se convirtió en un paso obligado hacia el sur de la provincia. Conoceremos también en este bloque la historia de la primer mujer intendente del departamento María Olga Said, quien fue derrocada por el golpe militar. Como espacio destacado encontramos la famosa Tragedia de los Arrieros. Luego, en el bloque 2 conoceremos algunos aspectos relevantes de la sociedad y cultura de Tupungato, la vida de José Fernandez, Pascual Perez, y las obras de unos artesanos conocidos internacionalemente. En el bloque 3 se destacan las características y alternativas de los turismos rurales de la zona, la producción vitivinícola, la tradicional vendimia departamental y conoceremos cuáles son los circuitos históricos-culturales más visitados de la zona.