The Spatial-Numerical Association of Response Codes (SNARC) effect in highly math-anxious individuals: An ERP study.

The Spatial-Numerical Association of Response Codes (SNARC) effect was examined in highly (HMA) and low math-anxious (LMA) individuals performing a number comparison in an ERP study. The SNARC effect consists of faster latencies when the response side is congruent with number location in the mental number line (MNL). Despite the stronger SNARC effect in the HMA group, their responses in incongruent trials were slower than in congruent trials only for the largest numerical magnitudes. Moreover, HMAs showed a less positive centroparietal P3b component in incongruent trials than in congruent ones, but only for the largest magnitudes. Since the SNARC effect arises during response selection and P3b positivity decreases with the difficulty of decision, this result suggests that HMA individuals might find it more difficult than LMAs to control the conflict between the automatically activated location of numbers in the MNL and the response side, especially in more cognitively demanding trials.

FAK phosphorylation plays a central role in thrombin-induced RPE cell migration.

The migration of retinal pigment epithelial (RPE) cells is an important step in various pathologic conditions including subretinal neovascularization (SRN), proliferative vitreoretinopathy (PVR) and, importantly, as a consequence of retinal surgery. Therefore, the elucidation of the mechanisms underlying RPE trans-differentiation and migration is essential for devising effective treatments aimed to the prevention of these disorders. A common event in these pathologies is the alteration of the blood-retina barrier (BRB), which allows the interaction of RPE cells with thrombin, a pro-inflammatory protease contained in serum. Our previous work has demonstrated that thrombin induces RPE cell cytoskeletal remodeling and migration, hallmark processes in the development of PVR; however, the molecular mechanisms involved are still unclear. Cell migration requires the disassembly of focal adhesions induced by Focal Adhesion Kinase (FAK) phosphorylation, together with the formation of actin stress fibers. The aim of the present work was to identify thrombin-activated signaling pathways leading to FAK phosphorylation and to determine FAK participation in thrombin-induced RPE cell migration. Results demonstrate that the activation of PAR1 by thrombin induces FAK autophosphorylation at Y397 and the subsequent phosphorylation of Y576/577 within the activation loop. FAK phosphorylation was shown to be under the control of c/nPKC and PI3K/PKC-ζ, as well as by Rho/ROCK, since the inhibition of these pathways prevented thrombin-induced FAK phosphorylation and the consequent disassembly of focal adhesions, in parallel to FAK-dependent actin stress fiber formation and RPE cell migration. These findings demonstrate, for the first time, that thrombin stimulation of RPE cell transformation and migration are regulated by FAK tyrosine phosphorylation. Thus, targeting FAK phosphorylation may provide a strategical basis for PVR treatment.

Mathematical anxiety effects on simple arithmetic processing efficiency: an event-related potential study.

This study uses event-related brain potentials to investigate the difficulties that high math anxious individuals face when processing dramatically incorrect solutions to simple arithmetical problems. To this end, thirteen high math-anxious (HMA) and thirteen low math-anxious (LMA) individuals were presented with simple addition problems in a verification task. The proposed solution could be correct, incorrect but very close to the correct one (small-split), or dramatically incorrect (large-split). The two groups did not differ in mathematical ability or trait anxiety. We reproduced previous results for flawed scores suggesting HMA difficulties in processing large-split solutions. Moreover, large-split solutions elicited a late positive component (P600/P3b) which was more enhanced and delayed in the HMA group. Our study proposes that the pattern of flawed scores found by previous studies (and that we replicate) has to do with HMA individuals'difficulties in inhibiting an extended processing of irrelevant information (large-split solutions).

Thrombin activation of PI3K/PDK1/Akt signaling promotes cyclin D1 upregulation and RPE cell proliferation.

The retinal pigment epithelium (RPE) plays an essential role in the survival and function of the neural retina. RPE uncontrolled proliferation leads to the development of proliferative ocular pathologies, among which proliferative vitreoretinopathy (PVR) is the main cause of retinal surgery failure. Upon the breakdown of the BRB due to trauma or metabolic imbalance the contact of RPE with serum-contained thrombin has been shown to stimulate the proliferation of otherwise quiescent RPE cells. Although the molecular mechanisms involved in this effect are still undetermined, thrombin proteolytic activation of protease-activated G protein coupled receptor-1 (PAR-1) activates PI3K and Akt, known to play an essential role in proliferation. The present study demonstrates that: 1) thrombin stimulates Ser 473 Akt phosphorylation without affecting Thr 308 basal phosphorylation in RPE cells; 2) thrombin-induced Akt stimulation promotes cyclin D1 accumulation through the phosphorylation/ inhibition of GSK-3ß, thus preventing Thr 286 cyclin D1 phosphorylation, nuclear export and degradation; 3) Akt signaling requires the upstream activation of PI3K and PLC. Since the pharmacological inhibition of these pathways or the silencing of cyclin expression prevent thrombin-induced RPE cell proliferation, these results contribute relevant evidence for establishing the mechanism involved in the development of proliferative eye diseases.

Developmental expression of N-methyl-D-aspartate glutamate receptor 1 splice variants in the chick retina.

Glutamate is the major excitatory neurotransmitter in the vertebrate retina. The N-methyl-D-aspartate glutamate receptor (NMDAR) is assembled as a tetramer containing NR1 and NR2, and possibly NR3 subunits, NR1 being essential for the formation of the ion channel. The NMDAR1 (NR1) gene encodes for mRNAs that generate at least eight functional variants by alternative splicing of exon 5 (cassette N1), 21 (cassette C1), or 22 (cassettes C2 or C2'). NR1 splice variants were identified in the mature chick retina, and their variation during embryonic development (ED) was analyzed. NR1 was shown to lack N1 in early ED, shifting to N1-containing variants in the mature retina, which could contribute to explaining the distinct biochemical properties of retinal NMDARs compared with the CNS. Sequence analysis of C-terminal variants containing C1 and C2 cassettes suggests a membrane-targeting mechanism for avian NMDARs distinct from that in mammals. An NR1 variant containing a novel alternative C-terminal splice exon named C3 was found, which encodes six amino acids containing a predicted casein kinase II phosphorylation site. This new variant is expressed in the retina during a restricted period of ED, coincident with the generation of spontaneous calcium activity waves, which precedes synapse formation in the retina, suggesting its participation in this process.

Red pigment-concentrating hormone induces a calcium-mediated retraction of distal retinal pigments in the crayfish.

The octapeptide red pigment-concentrating hormone is capable of eliciting the aggregation of intracellular pigment granules in distal retinal pigment cells of isolated retinas of the crayfish Procambarus clarkii (Girard). The final level and the time course of pigment aggregation are dose dependent within a range of 10(-10) mol l(-1) to 10(-4) mol l(-1). The effect of red pigment-concentrating hormone is prevented by previous incubation with an anti- red pigment-concentrating hormone antibody; however, application of the antibody after the onset of the red pigment-concentrating hormone effect, does not prevent its full development. A similar effect to that elicited by red pigment-concentrating hormone is induced by the calcium ionophores ionomycin and A-23187. Red pigment-concentrating hormone evokes entry of 45Ca2+ to retinal cells. However, the red pigment-concentrating hormone-induced pigment aggregation persists in the presence of the calcium channel blocker verapamil and in Ca2+-free solutions. Caffeine and thapsigargin, known to release calcium from intracellular stores, elicit distal pigment aggregation, while ryanodine and dantrolene, blockers of intracellular calcium release, as well as the intracellular calcium chelator bapta-AM suppress the effect of red pigment-concentrating hormone. These results suggest that red pigment-concentrating hormone elicits distal retinal pigment aggregation by increasing intracellular calcium concentration, acting via a dual mechanism: (1) promoting calcium entry, and (2) releasing intracellular calcium.

Glial transporters for glutamate, glycine, and GABA III. Glycine transporters.

Glial cells possess transport systems for the three major amino acid neurotransmitters glutamate, gamma-aminobutyric acid (GABA) and glycine, involved in the arrest of neurotransmission mediated by these compounds. Two glycine transporters have been cloned: GLYT1, mainly expressed by glial cells and shown to colocalize with NMDA receptors, and GLYT2, exclusively expressed by neurons and colocalized with the inhibitory glycine receptors. The way in which the regulation of extracellular glycine concentration by glial glycine transporters affects physiological and pathological conditions is discussed. The presence, differential pharmacology and specific regulation of glycine transporters in glial cells strongly support an important role for glia in the modulation of both, excitatory and inhibitory neurotransmission.

Glial transporters for glutamate, glycine and GABA I. Glutamate transporters.

The termination of chemical neurotransmission in the CNS involves the rapid removal of neurotransmitter from synapses by specific transport systems. Such mechanism operates for the three major amino acid neurotransmitters glutamate, gamma-aminobutyric acid (GABA) and glycine. To date, five different high-affinity Na(+)-dependent glutamate (Glu) transporters have been cloned: GLT1, GLAST, EAAC1, EAAT4 and EAAT5. The first two are expressed mainly by glial cells, and seem to be the predominant Glu transporters in the brain. A major function of Glu uptake in the nervous system is to prevent extracellular Glu concentrations from raising to neurotoxic levels in which glial transporters seem to play a critical role in protecting neurons from glutamate-induced excitotoxicity. Under particular conditions, glial GluTs have been shown to release Glu by reversal of activity, in a Ca(2+)--and energy-independent fashion. Furthermore, an activity of these transporters as ion channels or transducing units coupled to G-proteins has recently been reported. The localization, stoichiometry, and regulation of glial GluTs are outlined, as well as their possible contributions to nervous system diseases as ALS, AD and ischemic damage.

Glial transporters for glutamate, glycine, and GABA: II. GABA transporters.

The termination of chemical neurotransmission in the central nervous system (CNS) involves the rapid removal of neurotransmitter from synapses. This is fulfilled by specific transport systems in neurons and glia, including those for gamma-aminobutyric acid (GABA), the main inhibitory neurotransmitter in the brain. Glial cells express the cloned Na(+)/Cl(-)-dependent, high-affinity GABA transporters (GATs) GAT1, GAT2, and GAT3, as well as the low-affinity transporter BGT1. In situ hybridization and immunocytochemistry have revealed that each transporter shows distinct regional distribution in the brain and the retina. The neuronal vs. glial localization of the different transporters is not clear-cut, and variations according to species, neighboring excitatory synapses, and developmental stage have been reported. The localization, stoichiometry, and regulation of glial GATs are outlined, and the participation of these structures in development, osmoregulation, and neuroprotection are discussed. A decrease in GABAergic neurotransmission has been implicated in the pathophysiology of several CNS disorders, particularly in epilepsy. Since drugs which selectively inhibit glial but not neuronal GABA uptake exert anticonvulsant activity, clearly the establishment of the molecular mechanisms controlling GATs in glial cells will be an aid in the chemical treatment of several CNS-related diseases.

The influence of native-language phonology on lexical access: exemplar-Based versus abstract lexical entries.

This study used medium-term auditory repetition priming to investigate word-recognition processes. Highly fluent Catalan-Spanish bilinguals whose first language was either Catalan or Spanish were tested in a lexical decision task involving Catalan words and nonwords. Spanish-dominant individuals, but not Catalan-dominant individuals, exhibited repetition priming for minimal pairs differing in only one feature that is nondistinctive in Spanish (e.g.,[see text]), thereby indicating that they processed these words as homophones. This finding provides direct evidence both that word recognition uses a language-specific phonological representation and that lexical entries are stored in the mental lexicon as abstract forms.

[Comparison of the effectiveness and cost of treatment with humid environment as compared to traditional cure. Clinical trial on primary care patients with venous leg ulcers and pressure ulcers]. / Comparación de la efectividad y coste de la cura en ambiente húmedo frente a la cura tradicional. Ensayo clinico en pacientes de atención primaria con úlceras vasculares y por presión.

INTRODUCTION: The discovery of moist environment dressings as alternatives to the traditional treatments based on exposing wounds to air, opened new expectations for the care and treatment of chronic wounds. Over the years, these expectations have led to the availability of new moist environment dressings which have made it possible to improve the care provided to patients suffering this kind of wounds, as well as providing important reasons to weigh in terms of cost-benefit-effectiveness at the time of selecting which type of treatment should be employed. The lack of comparative analysis among traditional treatments and moist environment treatments for chronic wounds among patients receiving primary health care led the authors to perform an analysis comparing these aforementioned options of treatment on patients suffering venous leg ulcers or pressure ulcers. PATIENTS, MATERIALS AND METHODS: The authors designed a Randomized Clinical Trial involving patients receiving ambulatory care in order to compare the effectiveness and cost-benefit of traditional versus moist environment dressing during the treatment of patients suffering stage II or III pressure ulcers or venous leg ulcers. In this trial, variables related to effectiveness of both treatments, as well as their costs were analyzed. MAIN RESULTS: 70 wounds were included in this Randomized Clinical Trial, 41 were venous leg ulcers of which 21 received a moist environment treatment while 20 received traditional cure, the other 29 wounds were pressure ulcers of which 15 received moist environment dressings treatment and 14 received traditional dressings. No statistically significant differences were found among the defining variables for these lesions in either group under treatment. In the venous leg ulcer study group, the authors conclusions were an average of 18.13 days, 16.33 treatment sessions and a cost of 10,616 pesetas to heal one square centimeter of the initial surface area of a wound on patients treated with traditional treatment compared to an average of 18.22 days, 4.54 treatment sessions and a cost of 2409 pesetas to heal one square centimeter of the initial surface area of a wound on patients treated with moist environment dressings. In the pressure ulcers study group, the authors conclusions were an average of 12.18 days, 12.1 treatment sessions and a cost of 15,490 pesetas to heal one square centimeter of the initial surface area of a wound on patients treated with traditional treatment compared to an average of 7.12 days, 1.86 treatment sessions and a cost of 2610 pesetas to heal one square centimeter of the initial surface area of a wound on patients treated with moist environment dressings. COMMENTS: The results of this randomized clinical trial demosntrated that the moist environment treatment group was more effective and had a better cost-benefit ratio than the traditional treatment group in the treatment of pressure ulcers and venous leg ulcers on patients cared for by nursing personnel in primary health care centers all of which agrees with publications consulted by authors.

Calcium-independent release of [3H]spermine from chick retina.

Spermine has been shown to influence NMDA receptor function through an interaction at the coagonist site for glycine in the central nervous system (CNS) and the retina. In order to support a role for spermine as neurotransmitter or neuromodulator in the chick retina, specific stimulated-release of spermine should be demonstrated. Isolated chick retinas, preloaded with [3H]spermine, were stimulated with 1 mM NMDA and other glutamate agonists at ionotropic receptors, in a continuous superfusion system. [3H]spermine was released from the retina by depolarization with 50 mM KCl, in a Ca2+-independent manner. Inhibition of Na+/K+-ATPase by ouabain or digitoxigenin also induced spermine release following 36 min in the presence of the drugs; such effect seems unrelated to changes in Na+ electrochemical gradients, since nigericin and veratrine did not induce release in Na+ containing medium. The lack of effect of glutamate, NMDA and kainate at 1 mM concentration, suggests that release of spermine in the retina is mediated by the reversal of uptake and not necessarily linked to EAA-receptor activation.

Neocortical hyperexcitability after GABA withdrawal in vitro.

The sharp interruption of the intracortical instillation of exogenous gamma-aminobutyric acid (GABA), generates an epileptic focus in mammals. Seizures elicited by GABA withdrawal last several days or weeks. The present work reports that GABA withdrawal-induced hyperexcitability can be produced in vitro: a sudden withdrawal of GABA (5 mM; 120 min) or benzodiazepine (60 microM flunitrazepam) from the superfusion, induced a gradual increase in the amplitude of the evoked population spike (PS) recorded on neocortical slices. PS enhancement reached 150% above the control value 2.5 h after GABA withdrawal. GABA withdrawal-induced hyperexcitability was facilitated by progesterone. PS enhancement induced by GABA withdrawal was associated with an impairment of GABA transmission occurring before epileptiform discharges were fully established. Paired pulse inhibition and evoked [3H]-GABA release appear decreased; suggesting that cortical hyperexcitability as a result of GABA withdrawal involves pre-synaptic changes. Specific muscimol binding decreased during GABA superfusion but recovered after GABA withdrawal. However, the sensitivity of the post-synaptic response to 3alpha-OH-5alpha-pregnan-20-one or allopregnanolone (alloP) was enhanced after GABA withdrawal, suggesting a functional change in the GABA(A) receptors. The changes described may be the cellular correlates of the withdrawal syndromes appearing after interruption of the administration of GABA(A) receptor agonists.

[3H]Spermine binding to synaptosomal membranes from the chick retina.

The binding of [3H]spermine to synaptosomal membranes from chick retina was examined. Saturable specific binding of [3H]spermine to synaptosomal membranes from plexiform layers of retina (P1 and P2) has been characterized, and found to concentrate in the inner plexiform layer compared to the outer plexiform layer (Bmax=9.3 and 37 pmol/mg protein for P1 and P2, respectively). Kinetics of specific [3H]spermine binding yield a sigmoidal saturation curve, indicating positive cooperativity (nH: 2.4 and 3.2 for P1 and P2, respectively) with high affinity: Kapp=61 and 67 nM for P1 and P2. The time required to attain equilibrium at room temperature was less than 5 min in both fractions. Dose-response curves for spermine, spermidine, and diethylene-triamine (DET) show different potencies for inhibiting [3HDET. Our results support a role for polyamines (PA) as neurotransmitters or neuromodulators in the vertebrate retina.

The adenylate cyclase inhibitor MDL-12330A has a non-specific effect on glycine transport in Müller cells from the retina.

Müller glial cells express two transport systems for glycine (Gly): one with low affinity and another identified as GLYT1 with high affinity. The latter colocalizes with NMDA receptors in the CNS. Gly is considered as an obligatory coagonist at NMDA receptors, and, hence, the Gly transport system could contribute to the modulation of glutamate (Glu) excitatory transmission in the vertical pathways of the retina. For this reason, the regulation of Gly transport by cAMP was studied. We report here a non-specific effect of MDL-12330A, a compound reported to inhibit adenylate cyclase (AC), on Gly transport in Müller glia. This effect might be due to a toxic action on the cells, decreasing cell viability, and not to a specific inhibition of the adenylate cyclase. Non-specific effects of this drug should be considered when the participation of cAMP in any biological process is studied. We have clearly demonstrated that cAMP does not participate in the regulation of Gly transport in Müller glia.

Characterization of glycine transport in cultured Müller glial cells from the retina.

Rapid termination of the synaptic action of glutamate (Glu) and glycine (Gly) is achieved by uptake into the presynaptic terminal and glial cells. In the vertebrate CNS, Gly acts both as an inhibitory neurotransmitter and as a Glu modulator or coagonist at postsynaptic N-methyl-D-aspartate (NMDA) receptors. We have previously described NMDA receptors in Müller cells of chick retina coupled to the phosphoinositide cascade, the entry of calcium, and the activation of protein kinase C (PKC; López-Colomé et al. Glia 9:127-135, 1993). A colocalization of Gly transporters and NMDA receptors has been reported in brain tissue (Smith et al. Neuron 8:927-936, 1992); since the concentration of Gly could participate in the modulation of Glu excitatory transmission in the vertical pathways of the retina, transport of Gly in monolayer cultures of Müller cells was studied. Gly transport was found pH-sensitive with an optimum at pH 7.4. Kinetic analysis of the saturation curve for Gly within a concentration range of 0.01-2 mM, revealed two components of transport: a low-affinity system with Km = 1.7 mM, Vmax = 30 nmol/10 min/mg protein, and a high-affinity one with a Km = 27 microM, Vmax = 3 nmol/10 min/mg protein. Both systems were Na+ -dependent; the high-affinity system proved also dependent on external Cl- and was inhibited by sarcosine, characteristic of GLYT1 transporters. The inhibition of low-affinity uptake by 2-(methylamino)isobutyric acid (MeAIB) and 2-aminoisobutyric acid (AIB) suggests the presence of transport system A in Müller cells. The process is energy-requiring, since Gly transport was decreased by metabolic inhibitors. Data obtained are in keeping with a modulatory role for Müller glia on excitatory transmission in the retina.

Excitatory amino acid-induced inositol phosphate formation in cultured retinal pigment epithelium.

Excitatory amino acid (EAA)-induced production of inositolphosphates (IPs) was studied in primary cultures of chick retinal pigment epithelium (RPE) following in vitro incorporation of [3H] myo-inositol. Glutamic acid (L-glu) significantly increased [3H]-IPs accumulation (215%). L-glu agonists stimulated [3H]IPs accumulation in the following order of efficiency: N-methyl-D-aspartate (NMDA) > or = L-glu > quisqualate > or = kainate > (IS,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD). Stimulation was dependent on external Ca2+. The NMDA-induced response was blocked by (+)-5-methyl-10,11-dihydro-5H-dibenzo-cyclohepten-5,10-imine maleate (MK-801) and 3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP) and was decreased by the L-Ca2+-channel blockers verapamil and nifedipine as well as by dantrolene. The metabotropic glutamate receptor (mGluR) antagonist (+)-alpha-methyl-4-carboxyphenylglycine (+)MCPG inhibited 3,5-dihydroxyphenylglycine (DHPG) and ACPD-induced stimulation, which demonstrates the presence in RPE of mGluRs 1 and/or 5, as well as NMDA receptors coupled directly, or through the influx of external Ca2+, to phospholipase C activation. L-glu agonists showed no effect either on basal level of intracellular cyclic adenosine monophosphate, nor on forskolin- or carbachol-induced stimulation of adenylyl cyclase. Since L-glu is released from the retina upon illumination, and receptors for this compound are present in RPE, the activation of the inositide pathway could be involved in the regulation of retina-RPE interaction, which is essential for the visual process.

Spermine inhibits [3H]glycine binding at the NMDA receptors from plexiform layers of chick retina.

Saturable specific binding of glycine to synaptosomal membranes from plexiform layers of the retina has been described, which seems to correspond to the modulatory site on NMDA-receptors (26). Spermine inhibited specific [3H]glycine binding to membranes from synaptosomal fractions from the outer (P1) and the inner (P2) plexiform layers of 1-3 day-old chick retinas in a dose-dependent manner with an IC50 = 35 microM for the P1 fraction and 32 microM for the P2 fraction. Kinetic experiments and non-linear regression analysis of [3H]glycine-specific binding showed a Kd approximately 100-150 nM in both fractions, and a higher Bmax (4.11 +/- 0.47 pmol/mg protein) for the inner plexiform layer compared to the outer plexiform layer (Bmax = 2.76 +/- 0.25 pmol/mg protein). Strychnine-insensitive [3H]glycine binding was inhibited by 100 microM spermine, due to a reduction in Bmax (P1 = 0.84 +/- 0.16 pmol/mg protein; P2 = 0.81 +/- 0.16 pmol/mg protein) without affecting the Kd. Association and dissociation constants in the absence and presence of 50 microM spermine remained unchanged. Results demonstrate the presence of a single modulatory site for spermine on NMDA receptors, in both synaptic layers of the chick retina.

Strychnine-insensitive [3H] glycine binding to synaptosomal membranes from the chick retina.

The pharmacology and kinetics of strychnine-insensitive [3H] glycine binding to synaptic membranes from the outer (P1) and the inner (P2) plexiform layers of chick retina was studied. Inhibition curves for glycine, D-serine, 1-amincyclopropanecarboxylic acid (ACPC) and strychnine were analyzed by non-linear regression. Hill slopes for glycine and D-serine were not different from unity, whereas those for ACPC were < 1 in both fractions, revealing heterogeneity of binding sites in these membranes. Non-linear regression analysis of time course and saturation experiments strengthen the idea that [3H] glycine binds to more than one class of sites, with similar affinities at equilibrium. Antagonists of strychnine-insensitive glycine receptors in the CNS did not inhibit [3H] glycine binding to these membranes, which demonstrates that NMDA receptors in the retina have different structural requirements for ligand interaction at these sites. pH affected the specific binding, in agreement with the participation of specific amino acid residues at glycine binding sites on NMDA receptors, and also with functional studies in which the modulation of affinity at this site by protons has been observed. These results support previous studies regarding CPP and MK-801 binding, and provide evidence which indicates that the pharmacophore for glycine and other NMDA-related ligands is distinct for the retina, compared to the CNS, mainly regarding the effects of glycine-site antagonists.