Mejora de sensores vítreos sol-gel para la conservación preventiva de materiales históricos frente a la acidez

[ES] Los sensores a base de recubrimientos vítreos sol-gel dopados con ácido 2[4-(dimetil-amino) fenilazo] benzoico son capaces de cambiar su absorción óptica cuando se someten a distintas concentraciones de iones H3O+ y OH-. La respuesta de los sensores en ensayos de campo se estudió en Cracovia (Polonia) variando el procedimiento normal de uso, con el fin de mejorar su respuesta. Se midieron tanto los parámetros ópticos de los sensores como las condiciones ambientales (temperatura, humedad, presión y concentraciones de SO2 y de NOx). La respuesta de los sensores se analizó en términos de los cambios de su absorción visible. Dichos cambios se deben a reacciones locales de neutralización que tienen lugar en la superficie de los sensores, debido al efecto conjunto de los contaminantes de carácter ácido y a la humedad ambiental. Se establecieron correlaciones entre la concentración del contaminante principal (SO2) y la respuesta de los sensores para elaborar una calibración directa entre la absorción óptica y el pH ambiental. Los sensores pueden detectar y evaluar la acidez ambiental, así como alertar sobre la concentración de contaminantes ácidos que pueden dañar a la mayoría de los materiales históricos.[EN] Sensors based on sol-gel glassy coatings doped with 2[4-(dimethyl-amino) phenylazo] benzoic acid are able to change their optical absorption when they are submitted to different concentration of H3O+ and OH-. The sensors behaviour in field tests was studied in Cracow (Poland), varying the normal procedure of operation to improve their response. Both the sensors optical parameters and the environmental conditions (temperature, humidity, pressure, SO2 and NOx concentrations) were measured. The sensors response was analysed in terms of their visible absorbance changes, which are due to local neutralisation reactions in the sensors surface by the join effect of acid pollutants and humidity. Correlations between the main acid pollutant (SO2) concentration and the sensors response are established to provide a relation between the optical absorption and the environmental pH. The sensors are able to detect and monitorise environmental acidity, as well as to alert on the pollutant concentration that may damage most of the historical materials.The authors wish to acknowledge bilateral Polish-Spanish project Ref. PAN-CSIC 2003PL0011, European Marie Curie project Ref. MERG-CT-2004-516436 and Spanish project Ref. CICYT-MAT-2003-03231 for financing support. N.C. acknowledges CSIC-ESF for an I3P postdoctoral contract.Peer reviewe

Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTERS 1 and 2: fructose and xylitol/H+ symporters in pollen and young xylem cells

The genome of Arabidopsis thaliana contains six genes, AtPMT1 to AtPMT6 (Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTER 1–6), which form a distinct subfamily within the large family of more than 50 monosaccharide transporter-like (MST-like) genes. So far, only AtPMT5 [formerly named AtPLT5 (At3g18830)] has been characterized and was shown to be a plasma membrane-localized H+-symporter with broad substrate specificity. The characterization of AtPMT1 (At2g16120) and AtPMT2 (At2g16130), two other, almost identical, members of this transporter subfamily, are presented here. Expression of the AtPMT1 and AtPMT2 cDNAs in baker's yeast (Saccharomyces cerevisiae) revealed that these proteins catalyse the energy-dependent, high-capacity transport of fructose and xylitol, and the transport of several other compounds with lower rates. Expression of their cRNAs in Xenopus laevis oocytes showed that both proteins are voltage-dependent and catalyse the symport of their substrates with protons. Fusions of AtPMT1 or AtPMT2 with the green fluorescent protein (GFP) localized to Arabidopsis plasma membranes. Analyses of reporter genes performed with AtPMT1 or AtPMT2 promoter sequences showed expression in mature (AtPMT2) or germinating (AtPMT1) pollen grains, as well as in growing pollen tubes, hydathodes, and young xylem cells (both genes). The expression was confirmed with an anti-AtPMT1/AtPMT2 antiserum (αAtPMT1/2) raised against peptides conserved in AtPMT1 and AtPMT2. The physiological roles of the proteins are discussed and related to plant cell wall modifications

Kidins220/ARMS is an essential modulator of cardiovascular and nervous system development

The growth factor family of neurotrophins has major roles both inside and outside the nervous system. Here, we report a detailed histological analysis of key phenotypes generated by the ablation of the Kinase D interacting substrate of 220 kDa/Ankyrin repeat-rich membrane spanning (Kidins220/ARMS) protein, a membrane-anchored scaffold for the neurotrophin receptors Trk and p75NTR. Kidins220 is important for heart development, as shown by the severe defects in the outflow tract and ventricle wall formation displayed by the Kidins220 mutant mice. Kidins220 is also important for peripheral nervous system development, as the loss of Kidins220 in vivo caused extensive apoptosis of DRGs and other sensory ganglia. Moreover, the neuronal-specific deletion of this protein leads to early postnatal death, showing that Kidins220 also has a critical function in the postnatal brain

Malate transport by the vacuolar AtALMT6 channel in guard cells is subject to multiple regulation

Gas exchange in plants is controlled by guard cells, specialized cells acting as turgor pressure-driven valves. Malate is one of the major anions accumulated inside the vacuole during stomatal opening counteracting the positive charge of potassium. AtALMT6, a member of the aluminum-activated malate transporter family, is expressed in guard cells of leaves and stems as well as in flower organs of Arabidopsis thaliana. An AtALMT6-GFP fusion protein was targeted to the vacuolar membrane both in transient and stable expression systems. Patch-clamp experiments on vacuoles isolated from AtALMT6-GFP over-expressing Arabidopsis plants revealed large inward-rectifying malate currents only in the presence of micromolar cytosolic calcium concentrations. Further analyses showed that vacuolar pH and cytosolic malate regulate the threshold of activation of AtALMT6-mediated currents. The interplay of these two factors determines the AtALMT6 function as a malate influx or efflux channel depending on the tonoplast potential. Guard cell vacuoles isolated from Atalmt6 knock-out plants displayed reduced malate currents compared with wild-type vacuoles. This reduction, however, was not accompanied by phenotypic differences in the stomatal movements in knock-out plants, probably because of functional redundancy of malate transporters in guard cell vacuoles

The Arabidopsis central vacuole as an expression system for intracellular transporters: functional characterization of the Cl-/H+ exchanger CLC-7

Functional characterization of intracellular transporters is hampered by the inaccessibility of animal endomembranes to standard electrophysiological techniques. Here, we used Arabidopsis mesophyll protoplasts as a novel heterologous expression system for the lysosomal chlorideproton exchanger CLC-7 from rat. Following transient expression of a rCLC-7:EGFP construct in isolated protoplasts, the fusion protein efficiently targeted to the membrane of the large central vacuole, the lytic compartment of plant cells. Membrane currents recorded from EGFP-positive vacuoles were almost voltage independent and showed time-dependent activation at elevated positive membrane potentials as a hallmark. The shift in the reversal potential of the current induced by a decrease of cytosolic pH was compatible with a 2Cl-/1H+ exchange stoichiometry. Mutating the so-called gating glutamate into alanine (E245A) uncoupled chloride fluxes from the movement of protons, transforming the transporter into a chloride channel-like protein. Importantly, CLC-7 transport activity in the vacuolar expression system was recorded in the absence of the auxiliary subunit Ostm1, differently to recent data obtained in Xenopus oocytes using a CLC-7 mutant with partial plasma membrane expression. We also show that plasma membrane-targeted CLC-7E245A is non-functional in Xenopus oocytes when expressed without Ostm1. In summary, our data suggest the existence of an alternative CLC-7 operating mode, which is active when the protein is not in complex with Ostm1. The vacuolar expression system has the potential to become a valuable tool for functional studies on intracellular ion channels and transporters from animal cells