Use of morphological, biochemical and SRAP molecular markers to differentiate varieties of Cynara cardunculus L. (Asteraceae)

Dentro del complejo primario de Cynara cardunculus L. se encuentran diversas variedades botánicas: var. scolymus (alcaucil), var. altilis (cardo cultivado) y var. sylvestris (cardo silvestre). A lo largo de la historia del mejoramiento, la caracterización de los materiales ha evolucionado desde el uso de caracteres morfológicos hasta los modernos análisis moleculares, pasando por los marcadores bioquímicos. El objetivo del presente trabajo fue comparar la utilidad de los marcadores morfológicos, bioquímicos y moleculares para la caracterización de materiales pertenecientes a tres variedades botánicas de Cynara cardunculus. Tres cultivares de la var. scolymus, dos de la var. altilis y dos de la var. sylvestris fueron caracterizadas por variables morfovegetativas, proteínas de reserva y por marcadores moleculares a través de la técnica SRAP. Estas metodologías permitieron discriminar dos grupos: uno incluyendo las variedades de cardo cultivado y silvestre, y el otro, las variedades del alcaucil. Los datos moleculares y morfológicos permitieron además diferenciar los cultivares evaluados de la var. scolymus. Se concluye que los marcadores analizados son útiles para la caracterización intravarietal e intervarietal en programas de mejoramiento.In the Cynara cardunculus L. primary complex we can find a globe artichoke: Cynara cardunculus var. scolymus and other botanical varieties such as C. cardunculus var. altilis (cultivated cardoon) and C. cardunculus var. sylvestris (wild artichoke). Traditionally, they varieties were grouped according to morphological and biochemical traits and more recently, based on molecular markers. The aim of the present paper was to compare the use of morphological, biochemical and molecular markers to distinguish botanical varieties of Cynara cardunculus. Three accessions of var. scolymus, two accessions of var. altilis and two accessions of var. sylvestris were used. Different morphological variables, electrophoresis seed protein patterns by SDS-PAGE and SRAP markers were evaluated. The three kinds of markers grouped all materials in two clusters: one of them included the globe artichoke and the other the cardoons. SDS-PAGE is a valid technique for botanical varieties identification but the morphological and molecular data were more effective at discriminating botanical varieties and accessions within the scolymus group.Fil: Espósito, Andrea. Consejo Nacional de Investigaciones Científicas y TécnicasFil: Martin, Eugenia. Consejo Nacional de Investigaciones Científicas y TécnicasFil: Cravero, Vanina P.. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Genética y Mejoramiento Vegetal.Fil: Cointry, Enrique L.. Consejo Nacional de Investigaciones Científicas y Técnica

SEM and AFM studies of two-phase magnetic alkali borosilicate glasses

The morphology and composition of four types of two-phase alkali borosilicate glasses with magnetic atoms prepared by inductive melting have been studied. The results of scanning electron microscopy point to uniform distribution of Na, Si, and O atoms in these samples whilemagnetic iron atoms formball-shaped agglomerates. The magnetic properties of these agglomerates have been confirmed by magnetic force microscopy. Atomic force microscopy had shown that in these samples two different morphological structures, drop-like and dendrite net, are formed. The formation of dendrite-like structure is a necessary condition for production of porous magnetic glasses. The obtained results allowus to optimize the melting and heat treatment processes leading to production of porous alkali borosilicate glasses withmagnetic properties. The first results for nanocompositematerials on the basis ofmagnetic glasses containing the embedded ferroelectrics KH2PO4 demonstrate the effect of applied magnetic field on the ferroelectric phase transition. The morphology and composition of four types of two-phase alkali borosilicate glasses with magnetic atoms prepared by inductive melting have been studied. The results of scanning electron microscopy point to uniform distribution of Na, Si, and O atoms in these samples whilemagnetic iron atoms formball-shaped agglomerates. The magnetic properties of these agglomerates have been confirmed by magnetic force microscopy. Atomic force microscopy had shown that in these samples two different morphological structures, drop-like and dendrite net, are formed. The formation of dendrite-like structure is a necessary condition for production of porous magnetic glasses. The obtained results allowus to optimize the melting and heat treatment processes leading to production of porous alkali borosilicate glasses withmagnetic properties. The first results for nanocompositematerials on the basis ofmagnetic glasses containing the embedded ferroelectrics KH2PO4 demonstrate the effect of applied magnetic field on the ferroelectric phase transition.DAA

Functional human sperm capacitation requires both bicarbonate dependent-PKA activation and down-regulation of Ser/Thr phosphatases by Src family kinases

In all mammalian species studied so far, sperm capacitation correlates with an increase in protein tyrosine (Tyr) phosphorylation mediated by a bicarbonate-dependent cAMP/PKA pathway. Recent studies in mice revealed however that a Src Family Kinase (SFK) induced inactivation of serine/threonine (Ser/Thr) phosphatases is also involved in the signaling pathways leading to Tyr phosphorylation. In view of these observations and with the aim of getting a better understanding of the signaling pathways involved in human sperm capacitation, in the present work we investigated the involvement of both the cAMP/PKA and SFK/phosphatase pathways in relation to the capacitation state of the cells. For this purpose, different signaling events and sperm functional parameters were analyzed as a function of capacitation time. Results revealed a very early bicarbonate-dependent activation of PKA indicated by the rapid (1 min) increase in both phospho-PKA substrates and cAMP levels (p<0.05). However, a complete pattern of Tyr phosphorylation was detected only after 6 h-incubation at which time sperm exhibited the ability to undergo the acrosome reaction (AR) and to penetrate zona-free hamster eggs. Sperm capacitated in the presence of the SFK inhibitor SKI606 showed a decrease in both PKA substrate and Tyr phosphorylation levels which was overcome by exposure of sperm to the Ser/Thr phosphatase inhibitor okadaic acid (OA). However, OA was unable to induce phosphorylation when sperm were incubated under PKA-inhibitory conditions (i.e. in the absence of bicarbonate or presence of PKA inhibitor). Moreover, the increase in PKA activity by exposure to a cAMP analogue and a phosphodiesterase inhibitor did not overcome the inhibition produced by SKI606. Whereas the presence of SKI606 during capacitation produced a negative effect (p<0.05) on sperm motility, progesterone-induced AR and fertilizing ability, none of these inhibitions were observed when sperm were exposed to SKI606 and OA. Interestingly, different concentrations of inhibitors were required to modulate human and mouse capacitation revealing the species-specificity of the molecular mechanisms underlying this process. In conclusion, our results describe for the first time the involvement of both PKA activation and Ser/Thr phosphatase down-regulation in functional human sperm capacitation and provide convincing evidence that early PKA-dependent phosphorylation is the convergent regulatory point between these two signaling pathways.Fil: Battistone, Maria Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas -conicet. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Da Ros, Vanina Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas -conicet. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Salicioni, A.. University Of Massachussets; Estados UnidosFil: Navarrete, F.. University Of Massachussets; Estados UnidosFil: Krapf, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina. Universidad Nacional de Rosario; ArgentinaFil: Visconti, P. E.. University Of Massachussets;Fil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas -conicet. Instituto de Biología y Medicina Experimental (i); Argentin

The action of CGRP and SP on cultured skin fibroblasts

Calcitonin gene-related peptide (CGRP) is the most abundant neuropeptide in the skin, followed by substance P (SP), vasoactive intestinal peptide (VIP), and other neuropeptides in smaller amounts. the proliferative effect of neuropeptides on fibroblasts may affect wound healing and may be associated with hyperproliferative skin and mesenchymal disorders. Understanding the neuropeptidergic action on fibroblasts may provide relevant information to a deeper comprehension of the healing process. This study reviews the action of the main neuropeptides, CGRP and SP, on cultured human skin fibroblasts.A systematic literature search was conducted on Medline and Web of Science databases on December 21, 2013.A total of 74 articles were retrieved using the proposed search strategies and 3 were found in the references section of the selected articles. Thirteen of the retrieved articles studied the action of CGRP and SP on cultured human skin fibroblasts, 12 of which related to SP and 1 related to both CGRP and SP.Only one study was retrieved about the action of both CGRP and SP on cultured human skin fibroblasts. Further studies are necessary to investigate CGRP on skin fibroblasts and its role in the fibroplasia phase of wound healing.Universidade Federal de São Paulo Unifesp, Dept Surg, Plast Surg Div, Postgrad Program Translat Surg, BR-04024002 São Paulo, BrazilUniv Estadual Santa Cruz, Dept Agr & Environm Sci, BR-45662900 Ilheus, BA, BrazilUniv Estadual Santa Cruz, Dept Biol Sci, BR-45662900 Ilheus, BA, BrazilUniversidade Federal de São Paulo Unifesp, Dept Surg, Plast Surg Div, Postgrad Program Translat Surg, BR-04024002 São Paulo, BrazilWeb of Scienc

Endocannabinoids mediate hyposalivation induced by inflammogens in the submandibular glands and hypothalamus

Objective: The aim of this study was to investigate the factors that could participate on salivary glands hypofunction during inflammation and the participation of endocannabinoids in hyposalivation induced by the presence of inflammogens in the submandibular gland (SMG) or in the brain. Design: Salivary secretion was assessed in the presence of inflammogens and/or the cannabinoid receptor antagonist AM251 in the SMG or in the brain of rats. At the end of the experiments, some systemic and glandular inflammatory markers were measured and histopathological analysis was performed. Results: The inhibitory effect observed 1 h after lipopolysaccharide (LPS, 50 μg/50 μl) injection into the SMG (ig) was completely prevented by the injection of AM251 (5 μg/50 μl) by the same route (P < 0.05). The LPS (ig)-induced increase in PGE2 content was not altered by AM251 (ig), while the glandular production of TNFa induced by the endotoxin (P < 0.001) was partially blocked by it. Also, LPS injection produced no significant changes in the wet weight of the SMG neither damage to lipid membranes of its cells, nor significant microscopic changes in them, after hispopathological analysis, compared to controls. Finally, TNFα (100 ng/5 μl) injected intracerebro-ventricularly (icv) inhibited methacholine-induced salivary secretion evaluated 30 min after (P < 0.01), but the previous injection of AM251 (500 ng/5 μl, icv) prevented completely that effect. Conclusion: We conclude that endocannabinoids mediate the hyposialia induced by inflammogens in the SMG and in the brain. The hypofunction would be due to changes on signalling pathway produced by inflammatory compounds since anatomical changes were not observed. © 2013 Elsevier Ltd. All rights reserved.Fil: Prestifilippo, Juan Pablo. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Fisiología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Ciencias Biológicas. Cátedra de Fisiopatología; ArgentinaFil: Medina, Vanina Araceli. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Fisicomatemática. Cátedra de Física; ArgentinaFil: Mohn, Claudia Ester. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Fisiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Rodriguez, P. A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - La Plata. Unidad de Administración Territorial; Argentina. Universidad de Buenos Aires. Facultad de Odontología; ArgentinaFil: Elverdin, Juan Carlos. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Fisiología; ArgentinaFil: Fernández Solari, Jose Javier. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Fisiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentin

Utilisation de la très haute résolution spatiale pour la caractérisation des habitats de rongeurs, vecteurs de zoonoses à La Réunion

Connaître la distribution des espèces vectrices de zoonoses est fondamentale pour comprendre les dynamiques épidémiologiques et caractériser les zones à risque. Cette distribution peut être appréhendée à différentes échelles : régionalement, l'aire de répartition de ces espèces définit les limites d'occurrence de la transmission d'agents pathogènes et, localement, leurs niches déterminent l'aléa. Cette dernière échelle requiert une connaissance très précise des milieux qui peut être acquise par l'analyse d'images satellite à très haute résolution spatiale. Dans le cadre d'un projet de recherche sur la leptospirose à La Réunion (LeptOI), cette étude propose de mesurer le potentiel d'utilisation d'une image Pléiades (produit rééchantillonné à 50 cm) pour caractériser la distribution locale de différents petits mammifères terrestres autour de l'Etang de Saint-Paul. Trois campagnes de piégeage ont permis la capture de 134 animaux appartenant à quatre des cinq espèces présentes à La Réunion (Rattus rattus, R. norvegicus, Suncus murinus et Tenrec ecaudatus). Une analyse orientée-objet de l'image Pléiades a permis de discriminer dix classes d'occupation des sols avec une précision totale de 83,6%. Des indices paysagers ont été calculés à partir de cette classification autour de chacun des pièges (distances les plus courtes aux classes d&#8217;occupation, densité de contours et pourcentages de surface des classes au sein de zones tampon de 50 et de 100 mètres). Trois zones ("humide", "anthropisée" et "sèche") sont clairement discriminées par ces variables, qui pourront être utilisées comme déterminants écologiques de différents profils épidémiologiques : la zone humide étant propice à la leptospirose et la zone anthropisée étant un lieu de cooccurrence des quatre espèces et ainsi potentiellement une zone d'échanges de pathogènes

Multiple formin proteins participate in glioblastoma migration

BackgroundThe prognosis of glioblastoma remains poor, related to its diffuse spread within the brain. There is an ongoing search for molecular regulators of this particularly invasive behavior. One approach is to look for actin regulating proteins that might be targeted by future anti-cancer therapy. The formin family of proteins orchestrates rearrangement of the actin cytoskeleton in multiple cellular processes. Recently, the formin proteins mDia1 and mDia2 were shown to be expressed in glioblastoma in vitro, and their function could be modified by small molecule agonists. This finding implies that the formins could be future therapeutic targets in glioblastoma.MethodsIn cell studies, we investigated the changes in expression of the 15 human formins in primary glioblastoma cells and commercially available glioblastoma cell lines during differentiation from spheroids to migrating cells using transcriptomic analysis and qRT-PCR. siRNA mediated knockdown of selected formins was performed to investigate whether their expression affects glioblastoma migration.Using immunohistochemistry, we studied the expression of two formins, FHOD1 and INF2, in tissue samples from 93 IDH-wildtype glioblastomas. Associated clinicopathological parameters and follow-up data were utilized to test whether formin expression correlates with survival or has prognostic value.ResultsWe found that multiple formins were upregulated during migration. Knockdown of individual formins mDia1, mDia2, FHOD1 and INF2 significantly reduced migration in most studied cell lines. Among the studied formins, knockdown of INF2 generated the greatest reduction in motility in vitro. Using immunohistochemistry, we demonstrated expression of formin proteins FHOD1 and INF2 in glioblastoma tissues. Importantly, we found that moderate/high expression of INF2 was associated with significantly impaired prognosis.ConclusionsFormins FHOD1 and INF2 participate in glioblastoma cell migration. Moderate/high expression of INF2 in glioblastoma tissue is associated with worse outcome. Taken together, our in vitro and tissue studies suggest a pivotal role for INF2 in glioblastoma. When specific inhibiting compounds become available, INF2 could be a target in the search for novel glioblastoma therapies.Peer reviewe