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Biomedical subjects

A Peres

Publications and source records attributed to A Peres.

At least 19 recordsLinked to original sources
PubMed1992

Two currents activated by epidermal growth factor in EGFR-T17 fibroblasts.

Application of 10 nM Epidermal Growth Factor (EGF) to single EGFR-T17 fibroblasts induced a marked hyperpolarization that could last for tens of minutes; in many cases the first transient was followed by a series of oscillations of the membrane potential. The outward current responsible for the hyperpolarizing response could be recorded simultaneously to an increase in the intracellular calcium concentration, as measured with the fluorescent indicator fura-2. The conductance was nearly linear in the voltage range from -100 to +50 mV. While the EGF-induced current had many characteristics of a K+ current and was strongly reduced by 50 nM charybdotoxin (ChTx), its reversal potential was apparently more negative than the potassium equilibrium potential (VK). The application of 2 microM ouabain prior to EGF stimulation produced responses that were similar to those obtained without ouabain; however, under these conditions the EGF-induced current showed a reversal potential of -96.6 +/- 3.2 mV, very close to VK. Simultaneous application of both 2 microM ouabain and 50 nM ChTx completely abolished the response. It can be concluded that the response to EGF stimulation in EGFR-T17 cells consists of two components: the first is a current carried through Ca(2+)-activated K+ channels; the second is due to the acceleration of the operation of the Na+/K(+)-ATPase.

3T3 Cells↗
PubMed1992

P2Y purinoceptors in normal NIH 3T3 and in NIH 3T3 overexpressing c-ras.

The ability of purinergic agonists to induce Ca2+ responses has been tested in two lines of murine fibroblasts: normal NIH 3T3 fibroblasts and NIH 115.14, a clone expressing high levels [1] of the c-ras protooncogene. Both kinds of cells are responsive to ATP in the range 1 microM-1 mM; ADP and ATP gamma S are almost as potent as ATP, while AMP is unable to elicit a response. Ca2+ measurements performed in single cells by image analysis show great variability among cells but in each individual responding cell the Ca2+ rise occurs in an all-or-none fashion. The transient Ca2+ response does not depend on influx from the extracellular medium. Electrophysiological experiments reveal the activation of an outward current (at -50 mV) by ATP, probably due to Ca(2+)-activated K+ channels, confirming the absence of a substantial Ca2+ influx. Finally, stimulation by ATP produces a small but significant increase in the production of inositol phosphates. These results indicate that these cell lines possess purinergic receptors which are not integral membrane channels and which are coupled to InsP3 formation and may be therefore classified as P2Y.

3T3 Cells↗
PubMed1992

[Supraventricular tachycardia due to retrograde decremental conduction over accessory pathways].

PURPOSE: To discuss the clinical and the electrophysiologic findings, the differential diagnosis and the behaviour of concealed retrograde long conduction time accessory pathways. METHODS: Seventeen patients were submitted to electrophysiologic study using programmed electrical stimulation of the heart to assess the electrophysiologic properties of the accessory pathway. RESULTS: In all 17 studied patients, it was possible to advance the next atrial activation by giving a ventricular premature beat during the refractoriness of the His bundle. Of 17 patients, 7 developed signs and symptoms of heart failure (tachycardia-induced cardiomyopathy) due to the presence of incessant tachycardia. Eight patients were cured surgically and 1 underwent DC catheter ablation of the AV node. Six patients were successfully treated with antiarrhythmic drugs, one die of cancer and one still presents incessant tachycardia. CONCLUSION: The electrophysiologic study is essential for the differential diagnosis of the supraventricular tachycardias with a R-P' interval longer than P'-R interval where the incidence of tachycardiomyopathy is high in this group of patients. Surgery provides definitive cure of those patients leading to the regression of the signs and symptoms of heart failure.

Adolescent↗
PubMed1991

Characterization of the tyrosine phosphorylation of calpactin I (annexin II) induced by platelet-derived growth factor.

Stimulation in vivo of Swiss 3T3 fibroblasts with platelet-derived growth factor (PDGF) in the presence of orthovanadate induces the tyrosine phosphorylation of a 39 kDa protein, identified as the phosphorylated slow-migrating form of calpactin I (annexin II) heavy chain, p36. In fact, in PDGF-stimulated cells, anti-(calpactin I) antibodies recognize a doublet of bands, p36 and p39, and the latter disappears upon treatment with phosphatase. In many regards phosphorylation of p39 differs from the rapid and transient phosphorylation of the PDGF receptor and of other substrates: (a) it has slower kinetics but is then stable for longer periods of time; (b) it occurs at 37 degrees C but not at 4 degrees C; and (c) whereas most of the tyrosine-phosphorylated proteins are associated with membrane-enriched preparations, membrane association of p39 only occurs in the presence of Ca2+. Moreover, calpactin I leaks out of permeabilized cells at 0.1 microM free Ca2+, whereas it remains associated with the cells at concentrations of Ca2+ greater than or equal to 1 microM. PDGF does not stimulate phosphoinositide turnover (and thus Ca2+ mobilization) at 4 degrees C; thus it can be suggested that the Ca(2+)-dependent translocation of the protein to membrane/cytoskeletal structures is a necessary condition for its phosphorylation. In addition, calpactin I may not be a direct substrate for the PDGF receptor kinase, but rather the substrate of another tyrosine kinase activated by the receptor.

Animals↗
PubMed1991

Cytosolic calcium responses induced by photolytic release of 1,4,5-inositol trisphosphate in single human fibroblasts.

We have used the whole cell technique to microinject human fibroblasts with either 1,4,5-inositol trisphosphate (InsP3) or 'caged' InsP3, in order to study the mechanisms of transmembrane signalling related to mitogenic stimulations. Cytosolic Ca2+ elevations in response to 1,4,5 InsP3 diffusing from the patch pipette were difficult to detect, while 1,4,5 InsP3, photoreleased after loading the cell with its inactive precursor, was capable of generating not only a single cytosolic Ca2+ rise but sometimes triggered an oscillatory calcium response, similar to that often observed under mitogenic stimulation. We estimated that less than 100 nM InsP3 was sufficient to generate Ca2+ responses. The Ca2+ rise produced by the photoreleased InsP3 could fully activate the K+ channels present in the plasma membrane of human fibroblasts.

Calcium↗
PubMed1991

Characterization of Ca2+ transients induced by intracellular photorelease of InsP3 in mouse ovarian oocytes.

Ca2+ transients (measured with Fluo-3) were induced in single mouse ovarian oocytes by photolytic liberation of InsP3. The time course of cytosolic Ca2+ changes induced in this way is composed of distinct phases: upstroke, fast decline, slow declining plateau and fast decline to rest level. All the phases reflect mainly intracellular redistributions of the ion and not influx, since they are not strongly dependent on external Ca2+ or on changes in transmembrane potential. Often sustained Ca2+ oscillations followed the first InsP3-induced Ca2+ transient. These persisted for several minutes in the absence of external Ca2+. The initial rate of Ca2+ rise and the delay between the InsP3 stimulus and Ca2+ upstroke are correlated with the amount of liberated InsP3. A second InsP3 stimulation, applied during the plateau, causes only small Ca2+ elevations, lacking the upstroke phase. A second, full sized, transient could be elicited only after a complete return to the basal level. Vanadate, applied intracellularly, appeared to inhibit the re-uptake phase into the stores, stabilizing the plateau level. The present observations suggest that in mouse oocytes the InsP3-sensitive stores provide only a small and graded Ca2+ release which may then act as a trigger for a more substantial Ca(2+)-induced Ca2+ release (CICR) process.

Animals↗
PubMed1991

[Ca2+]i recordings and the inactivation of the high-voltage activated Ca2+ currents in the adult rat sensory neuron.

Fast, single cell, measurement of the average cytosolic [Ca2+]i with the Fura-2 technique suggests that the depolarization induced [Ca2+]i rise is entirely due to entry through the voltage-activated Ca2+ channels. Involvement of a Ca(2+)-induced Ca(2+)-release process is not evident. Under physiological cytosolic buffering the current-induced [Ca2+]i rise persists for seconds and decays exponentially (tau = 7 s). Analysis of the [Ca2+]i changes during two-pulse protocols indicates that the purely voltage-dependent inactivation of the high voltage-activated (HVA) channels, in the range -80/+70 mV, is a slow process (0.2-1 s) which removes at most 40% of the current. On the contrary, Ca(2+)-dependent inactivation acts in a fast way and it is therefore responsible for the fast inactivating phase of the current; this phase disappears under sustained [Ca2+]i loads, and reappears when redistribution of free Ca2+ takes place. A suitable correction may be devised to compensate for the Ca(2+)-dependent inactivation.

Animals↗
PubMed1991

Longitudinal dissociation of atrioventricular accessory pathways.

Unusual properties of atrioventricular (AV) accessory pathways were found during electrophysiologic investigations in four patients (three men and one woman). Anterograde longitudinal dissociation of the accessory pathway was observed in two patients and retrograde longitudinal dissociation in two others. Two patients had an accessory pathway with a slow conduction time, one in anterograde direction and one in retrograde direction. These observations further expand our knowledge of the spectrum of electrophysiologic properties of accessory AV pathways.

Adult↗
PubMed1990

InsP3- and Ca2(+)-induced Ca2+ release in single mouse oocytes.

To better understand the mechanism of intracellular Ca2+ mobilization, mouse oocytes were micro-injected with 'caged'-inositol-1,4,5 triphosphate caged-InsP3) together with the Ca2+ indicator Fluo-3 to directly induce and monitor Ca2+ redistribution. Photo-released InsP3 elicits [Ca2+]i changes exhibiting several kinetic phases and threshold behaviour. Often Ca2+ oscillations were induced after a single InsP3 pulse. Autoregenerative Ca2+ transients could also be induced by injections of Ca2+ itself, demonstrating unequivocally the presence of a Ca2(+)-induced Ca2(+)-release mechanism in these cells.

Animals↗
PubMed1990

Clinical and electrophysiologic characteristics of patients with antidromic circus movement tachycardia in the Wolff-Parkinson-White syndrome.

Antidromic circus movement tachycardia was documented in 36 of 345 consecutive patients with Wolff-Parkinson-White syndrome undergoing detailed electrophysiologic evaluation. Twenty-six patients were men and 10 were women (mean age +/- standard deviation 26 +/- 12 years [range 12 to 45]). Multiple accessory pathways were identified in 12 of these 36 patients (33%). Ten of the patients (67%) with clinically documented antidromic tachycardia had multiple accessory pathways. Dizziness and syncope occurred in 61 and 50% of patients with antidromic circus movement tachycardia. Six patients had clinical documentation of atrial fibrillation, and 4 patients (11%) were resuscitated from ventricular fibrillation. In the 36 patients, 56 distinct antidromic tachycardias were recorded and several different pathways were observed. Orthodromic tachycardia was the most frequently associated arrhythmia (72%). Dual atrioventricular nodal pathways were present in 12 patients (33%); however, atrioventricular nodal tachycardia could be initiated in only 2 of them. Interruption of the accessory pathway was successfully performed in all 20 patients undergoing surgery.

Adolescent↗
PubMed1990

Mitogen-induced oscillations of membrane potential and Ca2+ in human fibroblasts.

Using the whole-cell technique, we have measured recurring hyperpolarizations induced by fetal calf serum and bradykinin in human fibroblasts. By coupling fura-2 microfluorimetry to electrophysiology, we have also measured directly cytosolic Ca2+ and found that Ca2+ oscillations occur in synchrony with membrane currents. Mitogen stimulation of cells in which intracellular K+ had been replaced with Cs+ resulted in the abolishment of the outward current. We conclude then that the mitogen-induced recurring hyperpolarizations in human fibroblasts are due to the opening of Ca2(+)-activated K+ channels.

Benzofurans↗
PubMed1990

Cytosolic calcium and membrane conductance in response to platelet-derived growth factor and bradykinin stimulation in single human fibroblasts.

Bradykinin (BK) and platelet-derived growth factor (PDGF) act as mitogens and stimulate phosphatidylinositol (PI) turnover in human fibroblasts. By coupling whole-cell electrophysiological measurements with cytosolic Ca2+ determinations using fura-2 microfluorimetry, we have studied the changes in cytosolic calcium and in membrane conductance in single cells following stimulation with BK or PDGF. Both agonists produce variable patterns of response which include: single transient, sustained pulsations, damped oscillations, no response. In all cases, there is a very good temporal correlation between increases in intracellular Ca2+ and membrane current. The cytosolic calcium elevation appears to be insensitive to membrane potential changes, indicating that Ca2+ is released from an intracellular source. The Ca2(+)-activated current is not blocked by 1 microM apamin or by 0.5 mM (+)-tubocurarine; it is instead strongly reduced by 5 mM tetraethylammonium (TEA). We can conclude that BK and PDGF induce very similar early responses in human fibroblasts, and that the variable pattern of response does not depend on the particular mitogen used. The membrane currents are due to a kind of Ca2(+)-activated K+ channels which, according to their voltage-dependence and specific blockers, belong to the "maxi K+" class.

Bradykinin↗
PubMed1989

Voltage-dependent calcium current in adherent mouse 3T3 fibroblasts.

Whole-cell recording was performed on adherent mouse Swiss 3T3 fibroblasts. Depolarizations from a holding potential of -100 mV gave rise to a transient inward current. The voltage dependence, kinetic properties, and ionic selectivity of this current are identical to those described in the same cells kept in suspension after detachment from the culture dish [A. Pandiella, A. Malgaroli, J. Meldolesi, and L.M. Vicentini (1987) Exp. Cell. Res. 170, 175-185; W.H. Moolenaar, L.G.J. Tertoolen, and S.W. de Laat (1984) J. Biol. Chem. 259, 8066-8069].

Action Potentials↗
PubMed1989

Membrane conductance oscillations induced by serum in quiescent human skin fibroblasts.

1. Application of fetal calf serum to quiescent human fibroblasts, kept under whole-cell voltage clamp at positive potentials, induced a series of transient rises in membrane conductance. 2. The first transient increase in conductance developed with very short time lag (2-10 s) after serum addition, while the period between successive transients was 30-90 s, being remarkably constant in each particular cell. 3. Raising the Ca2(+)-buffering capacity of the intracellular solution with 1 mM-EGTA suppressed the appearance of the sustained oscillations. 4. The conductance increase was strongly voltage dependent: voltage ramps applied before, during and after the transients revealed the activation of an outwardly rectifying conductance with variable reversal potentials (between +14 and -55 mV). 5. No significant shifts of the reversal potential were observed when the extracellular K+ concentration was increased to 126 mM. Substitution of K+ with Cs+ as intracellular cation eliminated the outward current in response to serum. 6. External application of the Ca2+ ionophore A23187 elicited currents which were very similar in voltage dependence and time course to those triggered by serum. 7. The serum-induced response persisted unaffected by the absence of external Ca2+. The response was also seen in the presence of 1 mM-Cd2+ in the external solution. 8. Serum addition caused a rapid morphological rearrangement of the cells. 9. It is concluded that serum triggers a mobilization of Ca2+ from intracellular stores which in turn activates cationic channels.

Blood Physiological Phenomena↗
PubMed1989

The effects of polyinosinic:polycytidylic acid on the graft-versus-host reaction. III: Increased severity of the reaction with delayed pI:C treatment.

We have been investigating the effects of polyinosinic:polycytidylic acid (pI:C), an interferon inducer, on the graft-versus-host reaction. We have previously shown that pI:C treatment of C57BL/6xAF1 (B6AF1) recipient mice immediately before injection of C57BL/6 (B6) parental lymphocytes inhibited the immuno-suppression and pathological changes normally caused by the GVH reaction, by a mechanism apparently identical to that seen in F1 hybrid resistance (HR) to hematopoietic grafts. We now demonstrate that delaying pI:C treatment by as little as 48 hr produces the opposite effect. Treatment of recipient B6AF1 mice at different days after transfer of parental lymphocytes induced a marked increase in the severity of the GVH reaction, as measured by a decreased plaque-forming cell response to sheep erythrocytes; decreased proliferative response to the T and B cell mitogens PHA, Con A, and LPS; increased pathological changes in both lymphoid and nonlymphoid tissues; and increased GVH-associated mortality. This effect is unrelated to HR, as pI:C was able to augment the severity of the GVH reaction when A strain cells were injected into AxCBAF1 recipients, which do not manifest HR. Early pI:C treatment (1 and 2 days after parental cell transfer) increased the severity of the GVH reaction much more than later pI:C treatment (7 and 8 days after parental cell transfer). This observation, along with the demonstration of altered pathology in GVH mice treated with pI:C, suggests that the effect of pI:C is not mediated through a direct suppressive effect of IF on the cells responding in either the PFC or mitogen assays, but rather by the ability of IF to activate or suppress mechanisms involved in the development of GVH-induced alterations.

Animals↗
PubMed1988

Serum induces the immediate opening of Ca2+-activated channels in quiescent human fibroblasts.

Application of fetal calf serum to quiescent human fibroblasts produces an immediate (3-20 s delay) increase in membrane conductance which lasts about 20-30 s. This conductance is strongly outwardly-rectifying and has a reversal potential between -45 and -10 mV. The conductance increase may also be induced by application of the Ca2+ ionophore A23187 while it does not occur when intracellular K+ is replaced by Cs+. It is concluded that this early effect of serum is due to the opening of Ca2+-activated channels. This permeability change will alter the membrane potential and thus possibly interact with other voltage-sensitive processes induced by serum growth factors.

Blood↗
PubMed1988

A voltage-dependent calcium current in mouse Swiss 3T3 fibroblasts.

Patch-clamp experiments in the whole-cell mode have been performed in Swiss 3T3 mouse fibroblasts. Depolarizations from negative holding potential (Vh less than -60 mV) gave rise to a rapidly activating, fully inactivating, inward current of few tenths of nA in physiological saline at 35 degrees C. The current persisted when external Na+ was replaced by impermeant TMA+ and disappeared in 0 Ca2+, 1 mM EGTA. The current was reversible blocked by Co2+ and it was slightly reduced when external Ca2+ was substituted by Ba2+. Finally its reversal potential changed with Nernstian slope with increasing external Ca2+ concentrations. It is concluded that these cells possess a voltage-dependent Ca2+ channel.

Animals↗