Chemical composition of the small coastal lagoons of the Mediterranean Spanish littoral

About sixty small water bodies (coastal lagoons, marshes, salt pans, channels, springs, etc.) of the Spanish Mediterranean coast were sampled seasonally for one year (1979-1980), in order to study different aspects of their chemical composition. The concentrations of major ions (alkalinity, Cl-, Ca2+, Mg2+, Na+, and K+), nutrients (N.NO-3, N.NO2-, TRP and Si), oxygen and pH were determined for this purpose. The salt concentrations measured range between 0.4 and 361.3 g l-1. The samples have been divided into four classes of salinity (in g l-1): Cl, S < 5; C2, 5 40. Within these classes, the pattern of ionic dominance recorded is remarkably constant and similar to that found in most coastal lagoons (Cl- > So42- > Alk., for the anions, and Na+ > Mg2+ > Ca2+ > K+, for the cations), although other models occur especially in the first class. The dominance of Na+ and Cl-, as well as the molar ratios Mg2+/Ca2+ and Cl- / SO42- ,clearly increase from class Cl to class C4. The hyperhaline waters include different subtypes of the major brine type"c",, of EUGSTER & HARDIE (1978), the Na+ - (Mg2+) - Cl- - (SO42-) being the most frequent. Nutrient concentrations fall within a wide range (N.NO3 from 0.1 to 1100 mg-at 1-1; PRT from 0.01 to 23.56 mg-at l-1 and Si from 1.0 to 502.0 mg-at l-1). The oxygen values are very variable too, ranging between 0 and 14.4 ml l-1. Four different patterns of nutrient distribution have been distinguished based on the mean concentrations of N.NO3-, and TRP (mean values in mg-at l-1): A, N.NO3- < 10, TRP > l ; B, N.NO3- > 100, TRP < 1; C, 10 < N.NO3- < 100, TRP < 1; C, D, N.NO3- < 10, TRP < 1. As a rule, lagoons of low salinity (C1 and C2 classes) display the nutrient pattern C, and lagoons of high salinity (C3 and C4) show the nutrient pattern D. Model A only appears in waters of very low salinity, whereas model B does not seem to be related to salinity.

Structural organization of alpha-subunit from purified and microsomal toad kidney (Na+ + K+)-ATPase as assessed by controlled trypsinolysis

The membrane organization of the alpha-subunit of purified (Na+ + K+)-ATPase ((Na+ + K+)-dependent adenosine triphosphate phosphorylase, EC 3.6.1.3) and of the microsomal enzyme of the kidney of the toad Bufo marinus was compared by using controlled trypsinolysis. With both enzyme preparations, digestions performed in the presence of Na+ yielded a 73 kDa fragment and in the presence of K+ a 56 kDa, a 40 kDa and small amounts of a 83 kDa fragment from the 96 kDa alpha-subunit. In contrast to mammalian preparations (Jørgensen, P.L. (1975) Biochim. Biophys. Acta 401, 399-415), trypsinolysis of the purified amphibian enzyme led to a biphasic loss of (Na+ + K+)-ATPase activity in the presence of both Na+ and K+. These data could be correlated with an early rapid cleavage of 3 kDa from the alpha-subunit in both ionic conditions and a slower degradation of the remaining 93 kDa polypeptide. On the other hand, in the microsomal enzyme, a 3 kDa shift of the alpha-subunit could only be produced in the presence of Na+. Our data indicate that (1) purification of the amphibian enzyme with detergent does not influence the overall topology of the alpha-subunit but produces a distinct structural alteration of its N-terminus and (2) the amphibian kidney enzyme responds to cations with similar conformational transitions as the mammalian kidney enzyme. In addition, anti alpha-serum used on digested enzyme samples revealed on immunoblots that the 40 kDa fragment was better recognized than the 56 kDa fragment. It is concluded that the NH2-terminal of the alpha-subunit contains more antigenic sites than the COOH-terminal domain in agreement with the results of Farley et al. (Farley, R.A., Ochoa, G.T. and Kudrow, A. (1986) Am. J. Physiol. 250, C896-C906).

Lipoxygenase 2 and the early steps of jasmonic acid biosynthesis in "Arabidopsis thaliana"

Résumé Les oxylipines, telles que l'acide jasmonique (AJ ou jasmonate), jouent un rôle central en réponse à la blessure et à la pathogenèse. De nombreuses études ont montré l'importance de la voie canonique du jasmonate lors de la défense des plantes. De plus, un précurseur cyclopentenone de l'AJ, l'acide oxo-phyto-dienoic (OPDA), a été impliqué comme jouant le rôle d'une molécule signal lors de la défense contre certains pathogènes. En utilisant des mutants bloqués dans la biosynthèse de l'acide jasmonique (aos) ou dans sa perception (coi1-1), nous avons cherché à définir dans quelle mesure l'OPDA joue un rôle de signal induisant l'expression génétique en réponse à la blessure chez Arabidopsis. A l'aide de puces à ADN (microarray), nous avons montré que les transcriptomes d'aos et de coi1-1 sont très semblables après blessure, ce qui suggère que les produits d'AOS sont tous perçus via COI1. Pourtant, lorsqu'on analyse les métabolites présents chez ces mutants, une différence est visible, puisque aos n'accumule pas d'AJ, alors que coi1-1 en accumule encore rapidement après blessure. Nous avons étudié la possibilité qu'un mécanisme de régulation post-traductionnelle sur la voie de biosynthèse du jasmonate explique l'accumulation d'AJ chez coi1-1 après blessure. La lipoxygenase 2 (LOX2) est la première enzyme impliquée dans la biosynthèse de l'AJ et est donc une cible potentielle d'un tel mécanisme. Un indice sur la manière dont l'activité LOX pourrait être régulée vient du mutant fou2 (pour fatty acid oxygenation upregcilated 2) dans lequel l'activité LOX ainsi que le niveau d'AJ sont constitutivement élevés. Cette mutation implique un flux de cation dans la régulation de la production de l'AJ. De plus, il a été montré que plusieurs LOXs, dans des organismes autres que des plantes, peuvent lier le calcium. Nous montrons que l'activité LOX requiert l'addition de cations divalents pour être maximale in vitro, et que non seulement le calcium mais aussi le magnésium joue ce rôle. De plus, nous caractérisons un mutant récessif de LOX2 chez Arabidopsis (lox2-1). Ces plantes sont fertiles, et une analyse quantitative montre qu'elles accumulent toujours un peu d'AJ après blessure. Ceci suggère que LOX2 n'est pas la seule LOX impliquée dans la synthèse d'AJ. Aussi les plantes lox2-1 ne sont pas plus sensibles que les plantes de type sauvage lorsqu'elles sont infectées par la moisissure Botrytis cinerea ou lorsqu'elles sont exposées à un détritivore, néanmoins elles sont plus sensibles lorsqu'elles sont offertes en nourriture à un insecte herbivore. Les insectes et les plantes ont co-évolué conjointement, ainsi une plante ne contenant qu'un niveau réduit d'AJ favorise l'insecte. La disponibilité d'un mutant avec un niveau intermédiaire d'AJ va permettre de mieux comprendre pourquoi les plantes produisent autant de jasmonate. Abstract Oxylipins such as jasmonic acid (JA) play central roles in the wound response and during pathogenesis and many studies have confirmed the important role of the canonical jasmonate pathway in plant defense. Moreover, the cyclopentenone precursor of JA, oxo-phytodienoic acid (OPDA), is also thought to function as a signaling molecule in defense towards some pathogens. Its action was reported to depend on a different signal pathway to JA. By using mutants blocked in the biosynthesis (aos) or perception (coil-1) of JA, we investigated to which extend OPDA works as signaling molecule to trigger gene expression in the wound response of Arabidopsis. Using microarrays, we showed that aos and coil-1 transcriptome are similar in response to wounding, suggesting that products of AOS are all perceived by COI1. However, we found a difference between the two mutants at the metobolomic level, since aos is devoid of JA, but coil-1 can still rapidly accumulate JA upon wounding. We investigated the possibility that the post-translational activation of JA biosynthesis could explain the fast accumulation of JA in coil-1 plants upon wounding. Lipoxygenase (LOX) 2 is the first enzyme implicated in JA synthesis and was thus chosen as a potential target for posttranslational regulation. A clue as to how LOX activity might be regulated came from the fatty acid oxygenation upregulated 2 (foul) mutant in which LOX activity and JA levels are elevated. The foul mutant implicates cations flux in the regulation of JA production, and several LOXs in organisms other than plants have been shown to bind calcium. We showed that Arabidopsis LOX requires divalent cations for full activity in vitro, and that not only calcium but also magnesium can play this role. Moreover, a single recessive mutant of AtLOX2 was characterized. These plants are fully fertile. Quantitative oxylipin analysis showed that lox2-1 can still accumulate some JA after wounding, which suggests that LOX2 is not the only LOX involved in JA biosynthesis. lox2-1 plants do not show altered susceptibility to the fungus Botrytis cinerea or to a detritivore, however, they are more susceptible to an insect herbivore. The insect and plants are closely co-evolved and a reduced ability to synthesize JA favors the insect. The availability of a lox2-1 mutant with intermediate JA levels will further help understanding why plants produce elevated JA levels.

Effects of long-term irrigation with treated wastewater. Part I: Evolution of soil physico-chemical properties

The long-term impact of irrigation on a Mediterranean sandy soil irrigated with Treated wastewater (TWW) since 1980 was evaluated. The main soil properties (CEC, pH, size distribution, exchangeable cations and chloride, hydraulic conductivity) as well as the organic matter and Cu, Cr and Pb speciation in an irrigated soil and a non-irrigated control soil at various soil depths were monitored and compared during a 2 years experiment. In this first part, the evolution of the physico-chemical soil properties was described. The irrigation with TWW was beneficial with regard to water and nutrient supplying. All the exchangeable cations other than K(+) were higher in the irrigated soil than in the reference one. A part of the exchangeable cations was not fixed on the exchange complex but stored as labile salts or in concentrated soil solution. Despite the very sandy soil texture, both saturated and unsaturated hydraulic conductivity exhibited a significant diminution in the irrigated soil, but remained high enough to allow water percolation during rainy periods and subsequent leaching of accumulated salts, preventing soil salinization. In the irrigated soil, exchangeable sodium percentage (ESP) exhibited high values (20% on average) and the soil organic C was lower than in the reference. No significant effect was noticed on soil mineralogical composition due to irrigation. (C) 2010 Published by Elsevier Ltd.

Balanç hidroquímic d'una conca mediterrània en estat seminatural: el cas de la riera d'Arbúcies

A study of hydrochemical characteristics of the Arbúcies river (a seminatural mediterranean basin) has been undertaken during 1991. Mean solute yield for each ion has been determined using the correlation curves between discharge and concentration, and the flow duration curve. The hydrochemical budget of the Arbúcies drainage basin shows that the basin is a net source for al1 the elements. The cationic denudation rate obtained in this granitic basin is 5.24 keqhalyear. This high rate is due to the intensive chemical weathering of the granodionte, that delivers an important amount of cations which will release to be transported. The results demostrate clearly the strength of lithological control on stream loads, particulary on the dissolved component.

Topical and intravitreous administration of cationic nanoemulsions to deliver antisense oligonucleotides directed towards VEGF KDR receptors to the eye.

Antisense oligonucleotides (ODNs) specific for VEGFR-2-(17 MER) and inhibiting HUVEC proliferation in-vitro were screened. One efficient sequence was selected and incorporated in different types of nanoemulsions the potential toxicity of which was evaluated on HUVEC and ARPE19 cells. Our results showed that below 10 microl/ml, a 2.5% mid-chain triglycerides cationic DOTAP nanoemulsion was non-toxic on HUVEC and retinal cells. This formulation was therefore chosen for further experiments. In-vitro transfection of FITC ODNs in ARPE cells using DOTAP nanoemulsions showed that nanodroplets do penetrate into the cells. Furthermore, ODNs are released from the nanoemulsion after 48 h and accumulate into the cell nuclei. In both ex-vivo and in-vivo ODN stability experiments in rabbit vitreous, it was noted that the nanoemulsion protected at least partially the ODN from degradation over 72 h. The kinetic results of fluorescent ODN (Hex) distribution in DOTAP nanoemulsion following intravitreal injection in the rat showed that the nanoemulsion penetrates all retinal cells. Pharmacokinetic and ocular tissue distribution of radioactive ODN following intravitreal injection in rabbits showed that the DOTAP nanoemulsion apparently enhanced the intraretinal penetration of the ODNs up to the inner nuclear layer (INL) and might yield potential therapeutic levels of ODN in the retina over 72 h post injection.

Critical values of nitrogen indices in tomato plants grown in soil and nutrient solution determined by different statistical procedures

The objective of this study was to establish critical values of the N indices, namely soil-plant analysis development (SPAD), petiole sap N-NO3 and organic N in the tomato leaf adjacent to the first cluster (LAC), under soil and nutrient solution conditions, determined by different statistical approaches. Two experiments were conducted in randomized complete block design with four repli-cations. Tomato plants were grown in soil, in 3 L pot, with five N rates (0, 100, 200, 400 and 800 mg kg-1) and in solution at N rates of 0, 4, 8, 12 and 16 mmol L-1. Experiments in nutrient solution and soil were finished at thirty seven and forty two days after transplanting, respectively. At those times, SPAD index and petiole sap N-NO3 were evaluated in the LAC. Then, plants were harvested, separated in leaves and stem, dried at 70ºC, ground and weighted. The organic N was determined in LAC dry matter. Three statistical procedures were used to calculate critical N values. There were accentuated discrepancies for critical values of N indices obtained with plants grown in soil and nutrient solution as well as for different statistical procedures. Critical values of nitrogen indices at all situations are presented.

Direct Thy-1/alphaVbeta3 integrin interaction mediates neuron to astrocyte communication.

Thy-1 is an abundant neuronal glycoprotein of poorly defined function. We recently provided evidence indicating that Thy-1 clusters a beta3-containing integrin in astrocytes to induce tyrosine phosphorylation, RhoA activation and the formation of focal adhesions and stress fibers. To date, the alpha subunit partner of beta3 integrin in DI TNC1 astrocytes is unknown. Similarly, the ability of neuronal, membrane-bound Thy-1 to trigger astrocyte signaling via integrin engagement remains speculation. Here, evidence that alphav forms an alphavbeta3 heterodimer in DI TNC1 astrocytes was obtained. In neuron-astrocyte association assays, the presence of either anti-alphav or anti-beta3 integrin antibodies reduced cell-cell interaction demonstrating the requirement of both integrin subunits for this association. Moreover, anti-Thy-1 antibodies blocked stimulation of astrocytes by neurons but not the binding of these two cell types. Thus, neuron-astrocyte association involved binding between molecular components in addition to the Thy-1-integrin; however, the signaling events leading to focal adhesion formation in astrocytes depended exclusively on the latter interaction. Additionally, wild-type (RLD) but not mutated (RLE) Thy-1 was shown to directly interact with alphavbeta3 integrin by Surface Plasmon Resonance analysis. This interaction was promoted by divalent cations and was species-independent. Together, these results demonstrate that the alphavbeta3 integrin heterodimer interacts directly with Thy-1 present on neuronal cells to stimulate astrocytes.

Cation Transport Coupled to ATP Hydrolysis By the (Na, K)-ATPase : an integrated, animated model

An Adobe (R) animation is presented for use in undergraduate Biochemistry courses, illustrating the mechanism of Na+ and K+ translocation coupled to ATP hydrolysis by the (Na, K)-ATPase, a P-2c-type ATPase, or ATP-powered ion pump that actively translocates cations across plasma membranes. The enzyme is also known as an E-1/E-2-ATPase as it undergoes conformational changes between the E-1 and E-2 forms during the pumping cycle, altering the affinity and accessibility of the transmembrane ion-binding sites. The animation is based on Horisberger's scheme that incorporates the most recent significant findings to have improved our understanding of the (Na, K)-ATPase structure function relationship. The movements of the various domains within the (Na, K)-ATPase alpha-subunit illustrate the conformational changes that occur during Na+ and K+ translocation across the membrane and emphasize involvement of the actuator, nucleotide, and phosphorylation domains, that is, the "core engine" of the pump, with respect to ATP binding, cation transport, and ADP and P-i release.

A simple halide-to-anion exchange method for heteroaromatic salts and ionic liquids

A broad and simple method permitted halide ions in quaternary heteroaromatic and ammonium salts to be exchanged for a variety of anions using an anion exchange resin (A− form) in non-aqueous media. The anion loading of the AER (OH− form) was examined using two different anion sources, acids or ammonium salts, and changing the polarity of the solvents. The AER (A− form) method in organic solvents was then applied to several quaternary heteroaromatic salts and ILs, and the anion exchange proceeded in excellent to quantitative yields, concomitantly removing halide impurities. Relying on the hydrophobicity of the targeted ion pair for the counteranion swap, organic solvents with variable polarity were used, such as CH3OH, CH3CN and the dipolar nonhydroxylic solvent mixture CH3CN:CH2Cl2 (3:7) and the anion exchange was equally successful with both lipophilic cations and anions.

Heavy metal ions are potent inhibitors of protein folding.

Environmental and occupational exposure to heavy metals such as cadmium, mercury and lead results in severe health hazards including prenatal and developmental defects. The deleterious effects of heavy metal ions have hitherto been attributed to their interactions with specific, particularly susceptible native proteins. Here, we report an as yet undescribed mode of heavy metal toxicity. Cd2+, Hg2+ and Pb2+ proved to inhibit very efficiently the spontaneous refolding of chemically denatured proteins by forming high-affinity multidentate complexes with thiol and other functional groups (IC(50) in the nanomolar range). With similar efficacy, the heavy metal ions inhibited the chaperone-assisted refolding of chemically denatured and heat-denatured proteins. Thus, the toxic effects of heavy metal ions may result as well from their interaction with the more readily accessible functional groups of proteins in nascent and other non-native form. The toxic scope of heavy metals seems to be substantially larger than assumed so far.

Mid- to Late Holocene shoreline reconstruction and human occupation in Ancient Eretria (South Central Euboea, Greece)

Few studies have aimed to reconstruct landscape change in the area of Eretria (South Central Euboea, Greece) during the last 6000 years. The aim of this paper is to partially fill in this gap by examining the interaction be- tween Mid- to Late Holocene shoreline evolution and human occupation, which is documented in the area from the Late Neolithic to the Late Roman period (with discontinuities). Evidence of shoreline displacements is derived from the study of five boreholes (maximum depth of 5.25 m below the surface) drilled in the lowlands of Eretria. Based on sedimentological analyses and micro/macrofaunal identifications, different facies have been identified in the cores and which reveal typical features of deltaic progradation with marine, lagoonal, fluvio- deltaic and fluvial environments. In addition, a chronostratigraphy has been obtained based on 20 AMS 14C radio- carbon dates performed on samples of plant remains and marine/lagoonal shells found in situ. The main sequences of landscape reconstruction in the plain of Eretria can be summarized as follows: a marine environ- ment predominated from ca. 4000 to 3200 cal. BC and a gradual transition to shallow marine conditions is ob- served ca. 3200-3000 cal. BC due to the general context of deltaic progradation west of the ancient city. Subsequently, from ca. 3000 to 2000 cal. BC, a lagoon occupied the area in the vicinity of the Temple of Apollo and the settlement's development was restricted to several fluvio-deltaic levees, thus severely limiting human activities in the plain. From ca. 2000 to 800 cal. BC, a phase of shallow marine presence prevailed and constrained settlement on higher ground, forcing abandonment of the major part of the plain. Finally, since the eighth century BC, the sea has regressed southward and created the modern landscape.

Elastic properties and secondary structure formation of single-stranded DNA at monovalent and divalent salt conditions

Single-stranded DNA (ssDNA) plays a major role in several biological processes. It is therefore of fundamental interest to understand how the elastic response and the formation of secondary structures are modulated by the interplay between base pairing and electrostatic interactions. Here we measure force-extension curves (FECs) of ssDNA molecules in optical tweezers set up over two orders of magnitude of monovalent and divalent salt conditions, and obtain its elastic parameters by fitting the FECs to semiflexible models of polymers. For both monovalent and divalent salts, we find that the electrostatic contribution to the persistence length is proportional to the Debye screening length, varying as the inverse of the square root of cation concentration. The intrinsic persistence length is equal to 0.7 nm for both types of salts, and the effectivity of divalent cations in screening electrostatic interactions appears to be 100-fold as compared with monovalent salt, in line with what has been recently reported for single-stranded RNA. Finally, we propose an analysis of the FECs using a model that accounts for the effective thickness of the filament at low salt condition and a simple phenomenological description that quantifies the formation of non-specific secondary structure at low forces.

Soil Stabilization With Chemicals, Iowa Highway Research Board Bulletin No. 22, 1960

In the search for new soil stabilizing agents the effects of six organic cations on plastic limit, liquid limit, shrinkage limit, air-dry strength and rate of slaking of a highly plastic clay subsoil were studied. In all cases the plasticity index and shrinkage were reduced by the treatments. The air-dry strength was lowered in varying degree, which was the only undesirable effect noted. With one exception resistance to slaking was improved. It is concluded that large organic cations show promise as possible stabilizing agents for highly plastic fine-grained soils.

Intracellular thiol-mediated modulation of epithelial sodium channel activity.

The epithelial sodium channel ENaC is physiologically important in the kidney for the regulation of the extracellular fluid volume, and in the lungs for the maintenance of the appropriate airway surface liquid volume that lines the pulmonary epithelium. Besides the regulation of ENaC by hormones, intracellular factors such as Na(+) ions, pH, or Ca(2+) are responsible for fast adaptive responses of ENaC activity to changes in the intracellular milieu. In this study, we show that ENaC is rapidly and reversibly inhibited by internal sulfhydryl-reactive molecules such as methanethiosulfonate derivatives of different sizes, the metal cations Cd(2+) and Zn(2+), or copper(II) phenanthroline, a mild oxidizing agent that promotes the formation of disulfide bonds. At the single channel level, these agents applied intracellularly induce the appearance of long channel closures, suggesting an effect on ENaC gating. The intracellular reducing agent dithiothreitol fully reverses the rundown of ENaC activity in inside-out patches. Our observations suggest that changes in intracellular redox potential modulate ENaC activity and may regulate ENaC-mediated Na(+) transport in epithelia. Finally, substitution experiments reveal that multiple cysteine residues in the amino and carboxyl termini of ENaC subunits are responsible for this thiol-mediated inhibition of ENaC.