Systematic review and meta-analysis on coronary calcifications in COVID-19.

Chest CT is valuable to detect alternative diagnoses/complications of COVID-19, while its role for prognostication requires further investigation. Non-pulmonary radiological findings such as cardiovascular calcifications could increase the predictivity of clinical outcomes of COVID-19 patients beyond pulmonary involvement. Several observational studies have reported mixed results on the role of coronary calcifications in COVID-19 patients as a predictor of hospitalization, ventilatory support, and mortality. The purpose of the study is to systematically review the available evidence on the predictive role of cardiovascular calcifications in SARS-CoV2 disease. The meta-analysis confirms the prognostic significance of coronary calcifications on hospital mortality, and coronary calcifications (CAC ≠ 0) were associated with an OR for mortality of 2.19 (95% CI 1.36-3.52). CAC was neutral on respiratory outcomes, but it was associated with an increased trend of cardiovascular events. Coronary calcium appears as a promising biomarker imaging even in short-term outcomes (MACEs, hospital mortality) in a non-cardiovascular disease such as Sars-CoV2 infection. Further large studies are needed to confirm promising results of this imaging biomarker in non-cardiovascular disease.

Effect of compost application on the dynamics of carbon in a nectarine orchard ecosystem.

The aim of the present study was to compare the quantity and the type of carbon (C) stored during the 14-year lifetime of a commercial nectarine orchard ecosystem fertilized with mineral or organic fertilizers. The study was carried out in the Po valley, Italy, in a nectarine orchard of the variety Stark RedGold, grafted on GF677 hybrid peach × almond. Since orchard planting in August 2001, the following treatments were applied in a randomized complete block design with four replicates per block and compared: 1. unfertilized control; 2. mineral fertilization (including P and K at planting and N applied as NO3NH4 yearly at the rate of 70-130 kg ha-1); 3. compost application at a rate of 5 Mg DW ha-1 yr-1; 4. compost application at a rate of 10 Mg DW ha-1 yr-1. Compost was obtained from domestic organic wastes mixed with pruning material from urban ornamental trees and garden management after a 3-month stabilization period. Application of compost at the highest rate increased C in the soil; the amount of C sequestered was approximately 60% from amendment source and 40% from the net primary production of trees and grasses with a net increase of C compared to mineral fertilization. Compost application was found to be a win-win strategy to increase C storage in soil and, at the same time, to promote plant growth and yield to levels similar to those obtained with mineral fertilization. The rate of C application is crucial, indicated by the fact that compost supply at the rate of 10 Mg ha-1 yr-1 was the only fertilization strategy of the ones tested that resulted in higher C sequestration. This shows that compost amendment may stimulate an increase in the net primary production of plants.

Faulty neuronal determination and cell polarization are reverted by modulating HD early phenotypes.

Increasing evidence suggests that early neurodevelopmental defects in Huntington's disease (HD) patients could contribute to the later adult neurodegenerative phenotype. Here, by using HD-derived induced pluripotent stem cell lines, we report that early telencephalic induction and late neural identity are affected in cortical and striatal populations. We show that a large CAG expansion causes complete failure of the neuro-ectodermal acquisition, while cells carrying shorter CAGs repeats show gross abnormalities in neural rosette formation as well as disrupted cytoarchitecture in cortical organoids. Gene-expression analysis showed that control organoid overlapped with mature human fetal cortical areas, while HD organoids correlated with the immature ventricular zone/subventricular zone. We also report that defects in neuroectoderm and rosette formation could be rescued by molecular and pharmacological approaches leading to a recovery of striatal identity. These results show that mutant huntingtin precludes normal neuronal fate acquisition and highlights a possible connection between mutant huntingtin and abnormal neural development in HD.

Identification of two mutations in cis in the SCN1A gene in a family showing genetic epilepsy with febrile seizures plus (GEFS+) and idiopathic generalized epilepsy (IGE).

Mutations in the SCN1A gene causing either loss or gain of function have been frequently found in patients affected by genetic epilepsy with febrile seizures plus (GEFS+) or Dravet syndrome (also named severe myoclonic epilepsy in infancy SMEI). By mutation screening of the SCN1A gene, we identified for the first time a case of two missense mutations in cis (p.[Arg1525Gln;Thr297Ile]) in all affected individuals of an Italian family showing GEFS+ and idiopathic generalized epilepsy (IGE). The p.Arg1525Gln mutation was not previously reported yet and was predicted to be pathological by prediction tools, whereas the p.Thr297Ile was already identified in patients showing SMEI. Functional studies revealed that the Nav1.1 channels harboring both mutations were characterized by a significant shift in the activation curve towards more positive potentials. Our data demonstrate that the p.Arg1525Gln represents a novel mutation in the SCN1A gene altering the channel properties in the co-presence of the p.Thr297Ile.

Organic fertilization leads to increased peach root production and lifespan.

We evaluated the effects of mineral and organic fertilizers on peach root dynamics in the growing season from 2003 to 2006 in a nectarine (Prunus persica L.) orchard, planted in 2001 and located in the Po valley, northeastern Italy. Very few studies have conducted long-term investigations of root dynamics of fruit crops. Our main objective was to determine whether organic fertilizers affect root dynamics differently than mineral fertilizers. The experiment was a completely randomized block design with four replicates of three treatments: unfertilized, mineral fertilized and composted with municipal waste. Mineral fertilizers included P (100 kg ha(-1) year(-1)) and K (200 kg ha(-1) year(-1)) applied only at planting and N (70-130 kg ha(-1) year(-1)) split into two applications, one at 40 days after full bloom (60%) and the other in September (40%) each year. The compost fertilization represented a yearly rate of 10 metric tons (t) dry weight ha(-1), which approximates (in kg ha(-1) year(-1)) 240 N, 100 P and 200 K, split similarly to that described for the mineral fertilization of N. Both root growth and survival were evaluated at 20-day intervals during the growing season by the minirhizotron technique. Compost increased the production of new roots compared with the other treatments (P < 0.01). Roots were mainly produced at a depth of 41-80 cm and from March to May and in late summer. An analysis of covariance indicated no significant effect of soil nitrate on root production (P = 0.47). The root lifespan was longer in compost-treated trees than in mineral-fertilized or unfertilized trees (P < 0.01) and it was strongly affected by time of birth; roots born later in the summer lived longer than those born in the spring. Across years and treatments, the average root lifespan was positively correlated with soil nitrate (r = 0.60; P < 0.001). Variation in root lifespan with method of fertilization could be accounted for by variation in soil nitrate concentration as indicated by no effect of fertilizer treatment on root lifespan when soil nitrate was included as a covariate. These results reveal how shifting from mineral to organic fertilizers may shift both soil properties and nutrient availability, leading to changes in both root production and lifespan.

An optimized experimental strategy for efficient conversion of embryonic stem (ES)-derived mouse neural stem (NS) cells into a nearly homogeneous mature neuronal population.

NS cells are a homogeneous population of neural stem cells which were previously derived from embryonic stem cells as well as from the fetal and adult brain. Our previous reports have described a 21 day long neuronal differentiation protocol able to reproducibly convert adult SVZ-derived NS (aNS) cells into a population composed of 65% mature neurons and 35% glial cells. Here we have developed a different procedure specifically applicable to ES-derived NS cells in order to fully explore their neurogenic capacity. Differently from the aNS differentiation procedure, optimized neuronal output from ES-derived NS cells requires replating of the cells on appropriate substrates followed by sequential exposure to modified media. In these conditions, ES-derived NS cells differentiate into neurons with a barely appreciable quota of astrocytes and occasional oligodendrocytes. In particular, 21 days after the beginning of the treatment, 85% of the cells has differentiated into molecularly and electrophysiologically mature neurons belonging to the GABAergic lineage. The procedure, which is applicable with no considerable differences to different ES-derived NS cell lines and to NS cells at different passages, opens to the possibility of molecular and biochemical studies on close-to-uniform stem cell derived neurons.

Setting the conditions for efficient, robust and reproducible generation of functionally active neurons from adult subventricular zone-derived neural stem cells.

Although new culture conditions enable homogeneous and long-term propagation of radial glia-like neural stem (NS) cells in monolayer and serum-free conditions, the efficiency of the conversion of NS cells into terminally differentiated, functionally mature neurons is relatively limited and poorly characterized. We demonstrate that NS cells derived from adult mouse subventricular zone robustly develop properties of mature neurons when exposed to an optimized neuronal differentiation protocol. A high degree of cell viability was preserved. At 22 days in vitro, most cells (65%) were microtubule-associated protein 2(+) and coexpressed gamma-aminobutyric acid (GABA), GAD67, calbindin and parvalbumin. Nearly all neurons exhibited sodium, potassium and calcium currents, and 70% of them fired action potentials. These neurons expressed functional GABA(A) receptors, whereas activable kainate, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and N-methyl-D-aspartic acid receptors were present in approximately 80, 30 and 2% of cells, respectively. Antigenic and functional properties were efficiently and reliably reproduced across experiments and cell passages (up to 68). This is the first report showing a consistent and reproducible generation of large amounts of neurons from long-term passaged adult neural stem cells. Remarkably, the neuronal progeny carries a defined set of antigenic, biochemical and functional characteristics that make this system suitable for studies of NS cell biology as well as for genetic and chemical screenings.

Differentiating embryonic stem-derived neural stem cells show a maturation-dependent pattern of voltage-gated sodium current expression and graded action potentials.

A population of mouse embryonic stem (ES)-derived neural stem cells (named NS cells) that exhibits traits reminiscent of radial glia-like cell population and that can be homogeneously expanded in monolayer while remaining stable and highly neurogenic over multiple passages has been recently discovered. This novel population has provided a unique in vitro system in which to investigate physiological events occurring as stem cells lose multipotency and terminally differentiate. Here we analysed the timing, quality and quantity of the appearance of the excitability properties of differentiating NS cells which have been long-term expanded in vitro. To this end, we studied the biophysical properties of voltage-dependent Na(+) currents as an electrophysiological readout for neuronal maturation stages of differentiating NS cells toward the generation of fully functional neurons, since the expression of neuronal voltage-gated Na(+) channels is an essential hallmark of neuronal differentiation and crucial for signal transmission in the nervous system. Using the whole cell and single-channel cell-attached variations of the patch-clamp technique we found that the Na(+) currents in NS cells showed substantial electrophysiological changes during in vitro neuronal differentiation, consisting mainly in an increase of Na(+) current density and in a shift of the steady-state activation and inactivation curves toward more negative and more positive potentials respectively. The changes in the Na(+) channel system were closely related with the ability of differentiating NS cells to generate action potentials, and could therefore be exploited as an appropriate electrophysiological marker of ES-derived NS cells undergoing functional neuronal maturation.

A fast transient outward current in layer II/III neurons of rat perirhinal cortex.

The perirhinal cortex (PRC) is a supra-modal cortical area that collects and integrates information originating from uni- and multi-modal neocortical regions, transmits it to the hippocampus, and receives a feedback from the hippocampus itself. The elucidation of the mechanisms that underlie the specific excitable properties of the different PRC neuronal types appears as an important step toward the understanding of the integrative functions of PRC. In this study, we investigated the biophysical properties of the transient, I (A)-type K(+) current recorded in pyramidal neurons acutely dissociated from layers II/III of PRC of the rat (P8-P16). The current activated at about -50 mV and showed a fast monoexponential decay (tau(h) >> 14 ms at -30 to +10 mV). I (A) recovery from inactivation also had a monoexponential time course. No significant differences in the biophysical properties or current density of I (A) were found in pyramidal neurons from rats of different ages. Application of 4-AP (1-5 mM) reversibly and selectively blocked I (A), and in current clamp conditions it increased spike duration and shortened the delay of the first spike during repetitive firing evoked by sustained depolarizing current injection. These properties are similar to those of the I (A) found in thalamic neurons and other cortical pyramidal neurons. Our results suggest that I (A) contributes to spike repolarization and to regulate both spike onset timing and firing frequency in PRC neurons.

Design of surface properties of PET films: effect of fluorinated block copolymers.

This paper demonstrates that the addition of fluorinated block copolymers to PET solutions can be used to prepare PET films with controlled surface morphology, porosity and chemical composition, by exploiting the phenomenon known as breath figures (BF) formation during a spin-coating procedure. Surface features, such as number, depth and diameter of pores and chemical composition, can be tuned by varying the experimental conditions: relative humidity, solution composition and amount of the fluorinated block copolymer added to the PET solutions (in the range of 0.5-10 wt% with respect to PET). BF patterns are more evident at relatively high concentrations of PET (3 wt%) and content of fluorinated block copolymer (10 wt% with respect to PET) in the solution. According to the obtained results, the fluorinated block copolymer seems to play a role in different steps of the mechanism of BF formation. XPS measurements showed a surface composition much richer in fluorinated segments than expected from bulk composition. The combined surface roughness and surface segregation of fluorinated segments have only a limited effect on the macroscopic wettability of the surfaces.

Antibacterial activity of plastics coated with silver-doped organic-inorganic hybrid coatings prepared by sol-gel processes.

Silver-doped organic-inorganic hybrid coatings were prepared starting from tetraethoxysilane- and triethoxysilane-terminated poly(ethylene glycol)-block-polyethylene by the sol-gel process. They were applied as a thin layer (0.6-1.1 microm) to polyethylene (PE) and poly(vinyl chloride) (PVC) films and the antibacterial activity of the coated films was tested against Gram-negative (Escherichia coli ATCC 25922) and Gram-positive (Staphylococcus aureus ATCC 6538) bacteria. The effect of several factors (such as organic-inorganic ratio, type of catalyst, time of post-curing, silver ion concentration, etc.) was investigated. Measurements at different contact times showed a rapid decrease of the viable count for both tested strains. The highest antibacterial activity [more than 6 log reduction within 6 h starting from 106 colony-forming units (cfu) mL-1] was obtained for samples with an organic-inorganic weight ratio of 80:20 and 5 wt % silver salt with respect to the coating. For the coatings prepared by an acid-catalyzed process, a high level of permanence of the antibacterial activity of the coated films was demonstrated by repeatedly washing the samples in warm water or by immersion in physiological saline solution at 37 degrees C for 3 days. The release of silver ions per square meter of coating is very similar to that previously observed for polyamides filled with metallic silver nanoparticles; however, when compared on the basis of Ag content, the concentration of silver ions released from the coating is much higher than that released from 1 mm thick specimens of polyamide (PA) filled with silver nanoparticles. Transparency and good adhesion of the coating to PE and PVC plastic substrates without any previous surface treatment are further interesting features.

[Rotavirus diarrhea in a population covered by private health insurance in Córdoba, Argentina]. / Diarrea por rotavirus en una población hospitalaria privada de Córdoba (Argentina).

BACKGROUND: Rotavirus is the most common cause of severe diarrhea worldwide, especially in children under 5 years of age. OBJECTIVES: To study the incidence of rotavirus diarrhea in a population in Cordoba, Argentina. To determine signs and symptoms with potential predictive clinical diagnostic value and evaluate how rotavirus diarrhea affects length of hospital stay. MATERIAL AND METHODS: A case-control study was performed from April 1, 2002 to March 31, 2003. RESULTS: Seventy-three patients were included. There were 33 rotavirus (1) cases and 40 rotavirus (-) controls. The mean age of the rotavirus (1) patients was 16.8 6 2.3 months. There were 14 (42.4 %; 95 % CI: 27.2-59.2) girls and 19 (57.6 %; 95 % CI: 40.8-72.8) boys. The seasonal peak of the disease was observed in the fall (30.3 %; 95 % CI: 17.4-47.3) and at the beginning of the austral winter (39.3 %; 95 % CI: 24.7-56.3). The signs and symptoms associated with the infection were vomiting (OR 8.40; 95 % CI: 2.39-31.33) and dehydration (OR 3.73; 95 % CI: 1.06-13.81). In the 33 rotavirus (1) patients, the mean length of hospital stay was 2.6 days. CONCLUSIONS: One half of the hospitalized cases were produced by rotavirus. The seasonal peak was observed in autumn and winter (southern hemisphere). Rotavirus (1) patients more frequently presented severe vomiting and dehydration than rotavirus (-) patients, although length of hospital stay and diarrhea were longer in the control group.

Diarrea por rotavirus en una población hospitalaria privada de Córdoba (Argentina) / Rotavirus diarrhea in a population covered by private health insurance in Córdoba, Argentina

Antecedentes: Los rotavirus son la causa más común de diarrea grave, especialmente en menores de 5 años de edad. Objetivos: Conocer la incidencia de diarrea por rotavirus en una población hospitalaria privada de Córdoba (Argentina). Investigar los signos y síntomas que pudieran tener valor predictivo en el diagnóstico clínico de la enfermedad y evaluar cómo afecta la diarrea por rotavirus a la duración de la hospitalización. Material y métodos: Se planeó un estudio de casos y controles, realizado entre el primero de abril de 2002 y el 31 de marzo de 2003. Resultados: Se incluyeron 73 pacientes: 33 casos rotavirus positivos y 40 controles rotavirus negativos. La edad media de los pacientes rotavirus positivos fue de 16,8 ± 2,3 meses, 14 (42,4 %; intervalo de confianza del 95 % [IC 95 %]: 27,2-59,2) de sexo femenino y 19 (57,6 %; IC 95 %: 40,8-72,8) de sexo masculino. La mayoría de los casos se presentó a finales del otoño y principios del invierno austral (30,3 %; IC 95 %: 17,4-47,3) y (39,3 %; IC 95 %: 24,7-56,3). Los signos y síntomas clínicos que se asociaron de manera significativa a la infección fueron los vómitos (odds ratio [OR]: 8,40; IC 95 %: 2,39-31,33) y la deshidratación (OR: 3,73; IC 95 %: 1,06-13,81). Los 33 pacientes rotavirus positivos presentaron un promedio de 2,6 días de internación. Conclusiones: La mitad de los casos de diarrea en esta población hospitalizada fueron causados por rotavirus. Se presentan en otoño e invierno. Los pacientes infectados por rotavirus han tenido vómitos y deshidratación grave con mayor frecuencia que la observada en cuadros diarreicos en los que no se detectaron rotavirus en heces, sin embargo, los días de internación y los días de diarrea fueron significativamente más prolongados en el grupo de pacientes control

Background: Rotavirus is the most common cause of severe diarrhea worldwide, especially in children under 5 years of age. Objectives: To study the incidence of rotavirus diarrhea in a population in Cordoba, Argentina. To determine signs and symptoms with potential predictive clinical diagnostic value and evaluate how rotavirus diarrhea affects length of hospital stay. Material and methods: A case-control study was performed from April 1, 2002 to March 31, 2003. Results: Seventy-three patients were included. There were 33 rotavirus (+) cases and 40 rotavirus (­-) controls. The mean age of the rotavirus (+) patients was 16.8 6 2.3 months. There were 14 (42.4 %; 95 % CI: 27.2-59.2) girls and 19 (57.6 %; 95 % CI: 40.8-72.8) boys. The seasonal peak of the disease was observed in the fall (30.3 %; 95 % CI: 17.4-47.3) and at the beginning of the austral winter (39.3 %; 95 % CI: 24.7-56.3). The signs and symptoms associated with the infection were vomiting (OR 8.40; 95 % CI: 2.39-31.33) and dehydration (OR 3.73; 95 % CI: 1.06-13.81). In the 33 rotavirus (+) patients, the mean length of hospital stay was 2.6 days. Conclusions: One half of the hospitalized cases were produced by rotavirus. The seasonal peak was observed in autumn and winter (southern hemisphere). Rotavirus (+) patients more frequently presented severe vomiting and dehydration than rotavirus (-­) patients, although length of hospital stay and diarrhea were longer in the control group

Caveolin-1 expression and membrane cholesterol content modulate N-type calcium channel activity in NG108-15 cells.

Caveolins are the main structural proteins of glycolipid/cholesterol-rich plasmalemmal invaginations, termed caveolae. In addition, caveolin-1 isoform takes part in membrane remodelling as it binds and transports newly synthesized cholesterol from endoplasmic reticulum to the plasma membrane. Caveolin-1 is expressed in many cell types, including hippocampal neurons, where an abundant SNAP25-caveolin-1 complex is detected after induction of persistent synaptic potentiation. To ascertain whether caveolin-1 influences neuronal voltage-gated Ca2+ channel basal activity, we stably expressed caveolin-1 into transfected neuroblastoma x glioma NG108-15 hybrid cells [cav1(+) clone] that lack endogenous caveolins but express N-type Ca2+ channels upon cAMP-induced neuronal differentiation. Whole-cell patch-clamp recordings of cav1(+) cells demonstrated that N-type current density was reduced in size by approximately 70% without any significant change in the time course of activation and inactivation and voltage dependence. Moreover, the cav1(+) clone exhibited a significantly increased proportion of membrane cholesterol compared to wild-type NG108-15 cells. To gain insight into the mechanism underlying caveolin-1 lowering of N-current density, and more precisely to test whether this was indirectly caused by caveolin-1-induced enhancement of membrane cholesterol, we compared single N-type channel activities in cav1(+) clone and wild-type NG108-15 cells enriched with cholesterol after exposure to a methyl-beta-cyclodextrin-cholesterol complex. A lower Ca2+ channel activity was recorded from cell-attached patches of both cell types, thus supporting the view that the increased proportion of membrane cholesterol is ultimately responsible for the effect. This is due to a reduction in the probability of channel opening caused by a significant decrease of channel mean open time and by an increase of the frequency of null sweeps.

Attenuation of G protein-mediated inhibition of N-type calcium currents by expression of caveolins in mammalian NG108-15 cells.

1. Caveolins are integral proteins of glycolipid/cholesterol-rich plasmalemmal caveolae domains, where, they may function as a plasma membrane scaffold onto which many classes of signalling molecules, including receptors and heterotrimeric G proteins, can assemble. To ascertain whether caveolins influence G protein-mediated signal transduction, we stably expressed caveolin-1 and -3 isoforms in the neuroblastoma x glioma NG108-15 hybrid cell line, lacking endogenous caveolins. Subsequently, using whole-cell voltage clamp methods, we examined whether the modulation of N-type voltage-gated Ca2+ channels by G(o) protein-coupled, delta-type opioid receptors might be affected by recombinant caveolin expression. 2. In transfected NG108-15 cells, caveolins localized at the plasma membrane and, upon subcellular fractionation on sucrose density gradients, they co-localized in Triton-resistant, low buoyancy fractions, with endogenous G(o) protein alpha-subunits. 3. The voltage-dependent inhibition of omega-conotoxin GVIA-sensitive Ba2+ currents following either activation of delta-opioid receptors by the agonist [o-pen2,o-pen5]-enkephalin (DPDPE), or direct stimulation of G proteins with guanosine 5'-O-(thiotriphosphate) (GTPgammaS) was significantly attenuated in caveolin-expressing cells. The kinetics of Ca2+ channel inhibition were also modified by caveolins. 4. Overall, these results suggest that caveolins may negatively affect G protein-dependent regulation of voltage-gated N-type Ca2+ channels, presumably by causing a reduction of the available pool of activated G proteins.

ST14A cells have properties of a medium-size spiny neuron.

The ST14A cell line was previously derived from embryonic day 14 rat striatal primordia by retroviral transduction of the temperature-sensitive SV40 large T antigen. We showed that cell division and expression of nestin persists at 33 degrees C, the permissive temperature, whereas cell division ceases, nestin expression decreases, and MAP2 expression increases at the nonpermissive temperature of 39 degrees C. In this study, we further characterized the cells and found that they express other general and subtype-specific neuronal characteristics. ST14A cells express enolase and beta III-tubulin. Furthermore, they express the striatal marker DARPP-32, which is up-regulated upon differentiation of the cells by growth in serum-free medium. Stimulation with dopamine, the D2-dopamine receptor agonist quinpirole, or the D1-dopamine receptor agonist SKF82958 results in phosphorylation of CREB. Treatment of the cells with a mixture of reagents which stimulate the MAPK and adenylyl cyclase pathways radically changes the morphology of the ST14A cells. The cells develop numerous neurite-like appearing processes which stain with beta III-tubulin. Moreover, under these conditions, intracellular injection of rectangular depolarizing current stimuli elicits overshooting action potentials with a relatively fast depolarization rate when starting from a strongly hyperpolarized membrane potential. Taken together, these data imply that the ST14A cell line displays some of the characteristics of a medium-size spiny neuron subtype and provides a new tool to elucidate the pathways and molecules involved in medium-size spiny neuron differentiation and disease.

Role of motor imitation in traversability of surfaces by walking infants.

The ecological approach by Gibson stressed the role of visual cues in toddlers' locomotor activities. In particular, a rigid surface offers more traversability "affordances" to walkers (13-16 mo. old) than a deformable surface. On the other hand, imitation plays a relevant role in human behaviors. In our study a group of 17 walking infants (12-19 mo.) were asked to choose between locomoting on a rigid versus a deformable surface after having seen an older child walking on the deformable surface. A control group of 19 infants were asked to do the same task without having seen a model. The task was repeated three times. No relevant differences appeared between the two groups as to the choice of surfaces: the rigid surface was preferred by both groups. Some slight differences in the behavior of the experimental group were seen as a result of observing the model. Clearly, however, social stimulus. such as the sight of an older child performing a specific task, does not overcome the affordances which induce infants not to walk on the deformable surface.

Kinetic study of N-type calcium current modulation by delta-opioid receptor activation in the mammalian cell line NG108-15.

The voltage-dependent inhibition of N-type Ca2+ channel current by the delta-opioid agonist [D-pen2, D-pen5]-enkephalin (DPDPE) was investigated in the mammalian cell line NG108-15 with 10 microM nifedipine to block L-type channels, with whole-cell voltage clamp methods. In in vitro differentiated NG108-15 cells DPDPE reversibly decreased omega-conotoxin GVIA-sensitive Ba2+ currents in a concentration-dependent way. Inhibition was maximal with 1 microM DPDPE (66% at 0 mV) and was characterized by a slowing of Ba2+ current activation at low test potentials. Both inhibition and kinetic slowing were attenuated at more positive potentials and could be relieved up to 90% by strong conditioning depolarizations. The kinetics of removal of inhibition (de-inhibition) and of its retrieval (re-inhibition) were also voltage dependent. Both de-inhibition and re-inhibition were single exponentials and, in the voltage range from -20 to +10 mV, had significantly different time constants at a given membrane potential, the time course of re-inhibition being faster than that of de-inhibition. The kinetics of de-inhibition at -20 mV and of reinhibition at -40 mV were also concentration dependent, both processes becoming slower at lower agonist concentrations. The rate of de-inhibition at +80/+120 mV was similar to that of Ca2+ channel activation at the same potentials measured during application of DPDPE (approximately 7 ms), both processes being much slower than channel activation in controls (<1 ms). Moreover, the amplitude but not the time course of tail currents changed as the depolarization to +80/+120 mV was made longer. The state-dependent properties of DPDPE Ca2+ channel inhibition could be simulated by a model that assumes that inhibition by DPDPE results from voltage- and concentration-dependent binding of an inhibitory molecule to the N-type channel.

Action-potential-like depolarizations relieve opioid inhibition of N-type Ca2+ channels in NG108-15 cells.

The ability of action-potential-like waveforms (APWs) to attenuate opioid-induced inhibition of N-type Ca2+ channels was investigated in the neuroblastoma x glioma cell line NG108-15 using whole-cell voltage clamp methods. In in vitro differentiated NG108-15 cells, the opioid agonist [d-ala2]-methionine-enkephalin (DAME) reversibly decreased omega-conotoxin-GVIA-sensitive Ba2+ currents (N-type currents). Agonist-mediated inhibition of N-type currents could be transiently relieved by strong unphysiological depolarizing prepulses to +80 mV (facilitation). Significant facilitation was also achieved by conditioning the cell with a train of 15 APWs, which roughly mimicked physiological action potentials (1- to 6-ms-long depolarizations to +30 mV from a holding potential of -40 mV). The APW-induced facilitation depended on both conditioning pulse frequency and duration. Summation of the disinhibition produced by each APW was possible because reinhibition following repolarization to -40 mV was a much slower process (tau=88 ms) than the onset of facilitation at +80 mV (tau=7 ms). These results provide evidence that N-type Ca2+ channel facilitation may be a physiologically relevant process, and suggest that neuronal firing may relieve agonist-induced inhibition of N-type currents to an extent depending on both the shape of action potentials and the frequency of firing.