Drug Management of Epilepsy: Current Problems and the Possible Role of Calcium Antagonists

Prologue and Introduction Epilepsy is a disease which has struck fear into the hearts of both sufferers and onlookers for many centuries. Only in the past hundred years or so has effective medication become available, and medical management still relies on a small group of antiepileptic drugs (AEDs). These can often abolish seizures, and very frequently diminish their frequency such that patients can enjoy a normal life-style. A significant minority of patients, however, do not respond satisfactorily. Combining different drugs is complicated by their sharing the same side-effect - sedation - which seems more additive than does any therapeutic effect. It is for the benefit of these patients with refractory epilepsy that research continues, to improve our use of the present AEDs, and to find new drugs which might, when used alone or in combination, improve their lot. Therapeutic Drug Monitoring (TDM) In Chapter 1, current use of TDM in the epilepsy clinic is analysed. By recording physicians' decisions both before and after serum anticonvulsant concentrations were made available at 488 clinic visits, we found that management decisions were affected at 23% of these consultations. However, physicians did not appear to follow a "target concentration strategy" as a high proportion of results (26%) in the "target range" were followed by a change in dosage. A drawback with the "target concentration strategy" was highlighted by the correlation between carbamazepine concentration and time since dosing (P< 0.005). The possible benefits of an approach combining clinical and biochemical information are discussed. Cognitive function In Chapter 2, the effects of many AEDs on mental function are assessed. Deterioration in "cognitive" or "psychomotor" abilities is a generally recognised side-effect of all current AEDs. However, most evidence compares patients taking AEDs with healthy volunteers, the effect on mental function of the disease itself thus being uncontrolled. Other studies show short term deterioration in volunteers given AEDs for a limited period, or in patients abruptly taking an increased dose. These respectively fail to allow for any beneficial effect which controlling seizures might have on mental function, and the effect of tolerance to the drugs' side-effects. In EXPT. 2, 66 patients on AED therapy performed a battery of psychomotor function tests, and their results were compared with those of 14 untreated epileptic patients and 11 healthy controls. A clear "step-wise" deterioration in function was seen with reaction times, short-term memory, card-sorting, and finger-tapping speeds. Untreated epileptics fared worse than controls (P <0.05 - P <0.001) but better than treated patients (P< 0.05 -P <0.01). This demonstrated the deliterious effect of epilepsy itself. The drugs may aggravate this, though clearly the treated patients had more severe epilepsy. No differences were found between the individual drugs. In EXPT. 3, the effect of tolerance was demonstrated in a small group (n=13) of new patients commenced on carbamazepine. After an initial deterioration in reaction time (P<0.05) and finger-tapping (P< 0.001) at one week, these abilities returned to normal by twelve weeks, while mean serum concentrations of carbamazepine only fell from 8.5 mg/L to 7.1 mg/L. The relevance of short-term cognitive deterioration demonstrated in many studies is thus brought into question. Since diurnal variation in serum concentrations has been shown to correlate with carbamazepine neurotoxicity, EXPT. 4 tested the pharmacokinetics of a controlled-release preparation (Tegretol Retard, CBZ-CR). Eight healthy volunteers took this and conventional carbamazepine 200mg bd for two weeks in a double-blind, crossover fashion. Serum concentrations "plateaued" for 56h after single dose CBZ-CR, while chronic dosing resulted in diurnal fluctuation of only 12% compared with 24% on conventional carbamazepine (P <0.025), and produced less rapid changes in concentration (P< 0.02). Enzyme induction appeared similar with both preparations, but the bioavailability of CBZ-CR was possibly slightly lower. The "smoother" pharmacokinetic profile of CBZ-CR did not produce a detectable improvement in psychomotor function. Enzyme induction Many AEDs (carbamazepine, phenytoin, phenobarbitone) induce an increase in hepatic metabolising enzyme activity. This results in accelerated metabolism of the drugs themselves, of some other drugs which undergo oxidative metabolism, and of endogenous hormones. The clinical implications of this last aspect remain unclear. (Abstract shortened by ProQuest.)

A Complete Characterization of Maximal Symmetric Difference-Free families on {1,…\u3cem\u3en\u3c/em\u3e}.

Prior work in the field of set theory has looked at the properties of union-free families. This thesis investigates families based on a different set operation, the symmetricc difference. It provides a complete characterization of maximal symmetric differencefree families of subsets of {1, . . . n

Is Bauman's "liquid modernity" influencing the way we are doing science?

This commentary analyzes the possible effects of lightness—a typical attribute of modern (liquid) society, according to Bauman—on the way we are doing science. We share our opinion in an attempt to discern whether some unwanted practices that may affect our scientific results (such as technology misuse, bonus rewards, publishing under pressure, or indolence for getting accurate results) can be attributed, at least partially, to the liquid characteristic of modern society. We also examine whether the different systems that support science favor these actions, conspiring against what should be the primary goal of science: the search for truth. We finally consider several aspects that should be taken into account to rescue science from the intrusion of weightless actions.Fil: Mattiazzi, Ramona Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Vila Petroff, Martin Gerarde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentin

AKAP18δ Puts CaMKII in the Right Place at the Right Time: Implications for Cardiac Ca 2+ Handling

A delicate control of myocyte Ca2+ handling is essential for efficient excitation-contraction coupling (ECC) in the heart, and its alteration is associated with decreased contractility, arrhythmias, hypertrophy, and heart failure. During ECC, entry of Ca2+ from the extracellular space occurs through L-type Ca2+ channels and mediates Ca2+-dependent opening of RyR2 (ryanodine receptors) allowing massive movement of Ca2+ from the sarco-endoplasmic reticulum (SR) to the cytosol and ultimately triggering cell contraction. Thereafter, relaxation occurs primarily by the reuptake of Ca2+ into de SR by SERCA2a (SR Ca2+ ATPase 2a). Phosphorylation of proteins involved in Ca2+ cycling have critical functional consequences on ECC, including greater influx of Ca2+ through the L-type Ca2+ channels and a greater release of Ca2+ from the SR through RyR2 and a more efficient Ca2+ reuptake through SERCA2a as a result of phosphorylation of its regulatory protein PLN (phospholamban). Accumulating evidence indicates that spatial and temporal control of phosphorylation/dephosphorylation cycles are another crucial point of control of cardiac ECC. This control is achieved, at least in part, by a complex network of scaffolding, anchoring and adaptor proteins that recruit, compartmentalize, and regulate protein kinases in a location specific manner.1 AKAPs (A-kinase anchoring proteins) are the paradigm of this integrated regulatory system that have been extensively shown to coordinate spatially restricted cAMP-PKA (protein kinase A)-dependent signaling that provides a high level of specificity, contributing to adrenergic modulation of cardiomyocyte function.2 There are over 50 known AKAPs (including alternative-spliced forms) that target PKA to different sites within the cell. While AKAPs share their ability to bind PKA, they are remarkably diverse scaffolding proteins. Indeed, AKAPs couple PKA to different substrates, enhancing the rate and fidelity of their phosphorylation by the kinase. By bringing together different combinations of upstream and downstream signaling molecules, AKAPs provide the architectural infrastructure for specialization of the cAMP/PKA signaling network which is critical for the regulation of cardiac Ca2+ handling.Fil: Gonano, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Vila Petroff, Martin Gerarde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentin

TLR4 and NLRP3 Caspase 1- IL-1β- Axis are not Involved in Colon Ascendens Stent Peritonitis (Casp)-Associated Heart Disease

Hemodynamic collapse and myocardial dysfunction are among the major causes ofdeath in severe sepsis. The purpose of this study was to assess the role played by TLR4and by the NLRP3 inflammasome in the cardiac dysfunction that occurs after highgradepolymicrobial sepsis. We performed the colon ascendens stent peritonitis (CASP)surgery in Tlr4-/-, Nlrp3-/- and caspase-1-/- mice. We also assessed for the first time theelectrical heart function in the CASP model. The QJ interval was increased in wild-typeC57BL/6J mice after CASP when compared to sham controls, a result paralleled by anincrease in the cardiac action potential duration (APD). The decreases in ejectionfraction (EF), left-ventricle end diastolic volume (LVEDV), stroke volume, and cardiacoutput found after CASP were similar among all groups of mice. Similar heart responsewas found when Nlrp3-/- mice were submitted to high-grade CLP. Despite developingcardiac dysfunction similar to wild-types after CASP, Nlrp3-/- mice had reducedcirculating levels of IL-1β, IL-6 and TNF-α. Our results demonstrate that the geneticablation of Tlr4, Nlrp3, and caspase-1 does not prevent the cardiac dysfunction, despitepreventing the increase in pro-inflammatory cytokines, indicating that these are notfeasible targets to therapy in high-grade sepsis.Fil: López Alarcón, Maria Micaela. Universidade Federal do Rio de Janeiro; BrasilFil: Fernandez Ruocco, Maria Julieta. Universidade Federal do Rio de Janeiro; BrasilFil: Ferreira, Fabiano. Universidade Federal do Rio de Janeiro; BrasilFil: Paula Neto, Heitor A.. Universidade Federal do Rio de Janeiro; BrasilFil: Sepúlveda, Marisa Noemí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - la Plata. Centro de Investigaciones Cardiovasculares ; ArgentinaFil: Vila Petroff, Martin Gerarde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - la Plata. Centro de Investigaciones Cardiovasculares ; ArgentinaFil: Carvalho, Adriana Bastos. Universidade Federal do Rio de Janeiro; BrasilFil: Peroba Ramos, Isalira. Universidade Federal do Rio de Janeiro; BrasilFil: Branda, Hugo Justino. Universidade Federal do Rio de Janeiro; BrasilFil: Neto Paiva, Claudia. Universidade Federal do Rio de Janeiro; BrasilFil: Medei, Emiliano. Universidade Federal do Rio de Janeiro; Brasi

CaMKII-dependent ryanodine receptor phosphorylation mediates sepsis-induced cardiomyocyte apoptosis

Sepsis is associated with cardiac dysfunction, which is at least in part due to cardiomyocyte apoptosis. However, the underlying mechanisms are far from being understood. Using the colon ascendens stent peritonitis mouse model of sepsis (CASP), we examined the subcellular mechanisms that mediate sepsis-induced apoptosis. Wildtype (WT) CASP mice hearts showed an increase in apoptosis respect to WT-Sham. CASP transgenic mice expressing a CaMKII inhibitory peptide (AC3-I) were protected against sepsis-induced apoptosis. Dantrolene, used to reduce ryanodine receptor (RyR) diastolic sarcoplasmic reticulum (SR) Ca2+ release, prevented apoptosis in WTCASP. To examine whether CaMKII-dependent RyR2 phosphorylation mediates diastolic Ca2+ release and apoptosis in sepsis, we evaluated apoptosis in mutant mice hearts that have the CaMKII phosphorylation site of RyR2 (Serine 2814) mutated to Alanine (S2814A). S2814A CASP mice did not show increased apoptosis. Consistent with RyR2 phosphorylation-dependent enhancement in diastolic SR Ca2+ release leading to mitochondrial Ca2+ overload, mitochondrial Ca2+ retention capacity was reduced in mitochondria isolated from WT-CASP compared to Sham and this reduction was absent in mitochondria from CASP S2814A or dantrolene-treated mice. We conclude that in sepsis, CaMKII-dependent RyR2 phosphorylation results in diastolic Ca2+ release from SR which leads to mitochondrial Ca2+ overload and apoptosis.Fil: Sepúlveda, Marisa Noemí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Burgos, Juan Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Ciocci Pardo, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: González Arbeláez, Luisa Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Mosca, Susana Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Vila Petroff, Martin Gerarde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentin

Thyroid hormone disorder and the heart: The role of cardiolipin in calcium handling

New Findings: What is the central question of this study? Do alterations in thyroid status affect haemodynamic parameters and echocardiographic measurements in the rat postnatal heart, and calcium handling, contractility, relaxation and cardiolipin content in isolated rat cardiomyocytes? What is the main finding and its importance? An imbalance in phospholipids of the mitochondrial membrane such as cardiolipin is related to defects in mitochondrial function. T3-dependent cardiolipin signals contribute to the maintenance of mitochondrial homeostasis and involve Ca2+ handling, this pathway being more important in hypothyroidism. Abstract: The objective of this study was to evaluate whether alterations in thyroid status affect (1) haemodynamic parameters and echocardiographic measurements in the rat postnatal heart, and (2) calcium handling, contractility, relaxation and cardiolipin content in isolated rat cardiomyocytes. Sprague–Dawley rats aged 2 months treated with T3 (hyperthyroid, 20 μg/100 g body weight) or 0.02% methimazole (hypothyroid, w/v) for 28 days. Heart function was evaluated by echocardiography. Measurements of mean arterial pressure (MAP), heart rate, Ca2+ transients, cardiomyocyte shortening, number of spontaneous contractions per minute and cardiolipin (CL) content were performed. Thyroid disorders were associated with changes in pacemaker activity without modifications of MAP. Thyroid disorder induced changes in left ventricular diameter which were correlated with modifications of cardiac contractility (altered cell shortening and sarcoplasmic reticulum Ca2+ content). Endocrine disorders altered cardiomyocyte relaxation (reduction in the time to 50% re-lengthening and the time to 50% Ca2+ decay). Thyroid disorder increased the number of spontaneous contractions per minute (an index of pro-arrhythmogenic behaviour). CL content was increased only in hypothyroid rats. Changes in CL content, CL composition and CL–protein interaction in mitochondria from hypothyroid animals are responsible for alterations of contractile and relaxation cardiac function. This mechanism may be not be involved in T3-treated rats. Maintenance of euthyroidism is of crucial importance to preserve cardiac performance. An imbalance in relation to phospholipids of the mitochondrial membrane such as CL is related to defects in mitochondrial function. T3-dependent CL signals contribute to the maintenance of mitochondrial homeostasis and involve Ca2+ handling, this pathway being more important in hypothyroidism.Fil: D'Angelo, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Metabolismo del Fármaco. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Metabolismo del Fármaco; ArgentinaFil: Martinez, Candela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Metabolismo del Fármaco. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Metabolismo del Fármaco; ArgentinaFil: Arreche, Noelia Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Metabolismo del Fármaco. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Metabolismo del Fármaco; ArgentinaFil: Balaszczuk, Ana María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Metabolismo del Fármaco. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Metabolismo del Fármaco; ArgentinaFil: Fernandez, Maria del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Burgos, Juan Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Vila Petroff, Martin Gerarde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Fellet, Andrea L.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Metabolismo del Fármaco. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Metabolismo del Fármaco; Argentin

Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G

Background: The cGMP-dependent protein kinase G (PKG) phosphorylates the cardiac ryanodine receptor (RyR2) in vitro. We aimed to determine whether modulation of endogenous PKG alters RyR2-mediated spontaneous Ca2+ release and whether this effect is linked to a change in RyR2 phosphorylation. Methods: & Results: Human embryonic kidney (HEK293) cells with inducible RyR2 expression were treated with the cGMP analogue 8-Br-cGMP (100 μM) to activate endogenous PKG. In cells transfected with luminal Ca2+ sensor, D1ER, PKG activation significantly reduced the threshold for RyR2-mediated spontaneous Ca2+ release (93.9 ± 0.4% of store size with vehicle vs. 91.7 ± 0.8% with 8-Br-cGMP, P = 0.04). Mutation of the proposed PKG phosphorylation sites, S2808 and S2030, either individually or as a combination, prevented the decrease in Ca2+ release threshold induced by endogenous PKG activation. Interestingly, despite a functional dependence on expression of RyR2 phosphorylation sites, 8-Br-cGMP activation of PKG did not promote a detectable change in S2808 phosphorylation (P = 0.9). Paradoxically, pharmacological inhibition of PKG with KT 5823 (1 μM) also reduced the threshold for spontaneous Ca2+ release through RyR2 without affecting S2808 phosphorylation. Silencing RNA knockdown of endogenous PKG expression also had no quantifiable effect on RyR2 S2808 phosphorylation (P = 0.9). However, unlike PKG inhibition with KT 5823, PKG knockdown did not alter spontaneous Ca2+ release propensity or luminal Ca2+ handling. Conclusion: In an intact cell model, activation of endogenous PKG reduces the threshold for RyR2-mediated spontaneous Ca2+ release in a manner dependent on the RyR2 phosphorylation sites S2808 and S2030. This study clarifies the regulation of RyR2 Ca2+ release by endogenous PKG and functionally implicates the role of RyR2 phosphorylation.Fil: Gonano, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentina. University of Otago; Nueva ZelandaFil: Aitken Buck, Hamish M.. University of Otago; Nueva ZelandaFil: Chakraborty, Akash D.. University of Otago; Nueva ZelandaFil: Worthington, Luke P. I.. University of Otago; Nueva ZelandaFil: Cully, Tanya R.. University of Otago; Nueva ZelandaFil: Lamberts, Regis R.. University of Otago; Nueva ZelandaFil: Vila Petroff, Martin Gerarde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Jones, Peter P.. University of Otago; Nueva Zeland

Aldosterone stimulates the cardiac sodium/bicarbonate cotransporter via activation of the g protein-coupled receptor gpr30

Some cardiac non-genomic effects of aldosterone (Ald) are reported to be mediated through activation of the classic mineralocorticoid receptor (MR). However, in the last years, it was proposed that activation of the novel G protein-coupled receptor GPR30 mediates certain non-genomic effects of Ald. The aim of this study was to elucidate if the sodium/bicarbonate cotransporter (NBC) is stimulated by Ald and if the activation of GPR30 mediates this effect. NBC activity was evaluated in rat cardiomyocytes perfused with HCO3-/CO2 solution in the continuous presence of HOE642 (sodium/hydrogen exchanger blocker) during recovery from acidosis using intracellular fluorescence measurements. Ald enhanced NBC activity (% of δJHCO3-; control: 100±5.82%, n=7 vs Ald: 151.88±11.02%, n=5; P<0.05), which was prevented by G15 (GPR30 blocker, 90.53±7.81%, n=7). Further evidence for the involvement of GPR30 was provided by G1 (GPR30 agonist), which stimulated NBC (185.13±18.28%, n=6; P<0.05) and this effect was abrogated by G15 (124.19±10.96%, n=5). Ald- and G1-induced NBC stimulation was abolished by the reactive oxygen species (ROS) scavenger MPG and by the NADPH oxidase inhibitor apocynin. In addition, G15 prevented Ald- and G1-induced ROS production. Pre-incubation of myocytes with wortmannin (PI3K-AKT pathway blocker) prevented Ald- or G1-induced NBC stimulation. In summary, Ald stimulates NBC by GPR30 activation, ROS production and AKT stimulation.Fil: de Giusti, Verónica Celeste. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Orlowski, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Ciancio, Maria Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Espejo, María Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Gonano, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Caldiz, Claudia Irma. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Vila Petroff, Martin Gerarde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Villa-Abrille, María Celeste. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Aiello, Ernesto Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentin