To explore whether CaMKII-dependent phosphorylation events mediate reperfusion arrhythmias, Langendorff perfused hearts were submitted to global ischemia/reperfusion. Epicardial monophasic or transmembrane action potentials and contractility were recorded. In rat hearts, reperfusion significantly increased the number of premature beats (PBs) relative to pre-ischemic values. This arrhythmic pattern was associated with a significant increase in CaMKII-dependent phosphorylation of Ser2814 on Ca2+-release channels (RyR2) and Thr17 on phospholamban (PLN) at the sarcoplasmic reticulum (SR). These phenomena could be prevented by the CaMKII-inhibitor KN-93. In transgenic mice with targeted inhibition of CaMKII at the SR membranes (SR-AIP), PBs were significantly decreased from 31 ± 6 to 5 ± 1 beats/3 min with a virtually complete disappearance of early-afterdepolarizations (EADs). In mice with genetic mutation of the CaMKII phosphorylation site on RyR2 (RyR2-S2814A), PBs decreased by 51.0 ± 14.7 %. In contrast, the number of PBs upon reperfusion did not change in transgenic mice with ablation of both PLN phosphorylation sites (PLN-DM). The experiments in SR-AIP mice, in which the CaMKII inhibitor peptide is anchored in the SR membrane but also inhibits CaMKII regulation of L-type Ca2+ channels, indicated a critical role of CaMKII-dependent phosphorylation of SR proteins and/or L-type Ca2+ channels in reperfusion arrhythmias. The experiments in RyR2-S2814A further indicate that up to 60% of PBs related to CaMKII are dependent on the phosphorylation of RyR2 Ser2814 site and could be ascribed to delayed-afterdepolarizations (DADs). Moreover, phosphorylation of PLN-Thr17 and L-type Ca2+ channels might contribute to reperfusion-induced PBs, by increasing SR Ca2+ content and Ca2+ influx.

Returning to normal pH after acidosis, similar to reperfusion after ischemia, is prone to arrhythmias. The type and mechanisms of these arrhythmias have never been explored and were the aim of the present work. Langendorff-perfused rat/mice hearts and rat-isolated myocytes were subjected to respiratory acidosis and then returned to normal pH. Monophasic action potentials and left ventricular developed pressure were recorded. The removal of acidosis provoked ectopic beats that were blunted by 1 muM of the CaMKII inhibitor KN-93, 1 muM thapsigargin, to inhibit sarcoplasmic reticulum (SR) Ca(2+) uptake, and 30 nM ryanodine or 45 muM dantrolene, to inhibit SR Ca(2+) release and were not observed in a transgenic mouse model with inhibition of CaMKII targeted to the SR. Acidosis increased the phosphorylation of Thr(17) site of phospholamban (PT-PLN) and SR Ca(2+) load. Both effects were precluded by KN-93. The return to normal pH was associated with an increase in SR Ca(2+) leak, when compared with that of control or with acidosis at the same SR Ca(2+) content. Ca(2+) leak occurred without changes in the phosphorylation of ryanodine receptors type 2 (RyR2) and was blunted by KN-93. Experiments in planar lipid bilayers confirmed the reversible inhibitory effect of acidosis on RyR2. Ectopic activity was triggered by membrane depolarizations (delayed afterdepolarizations), primarily occurring in epicardium and were prevented by KN-93. The results reveal that arrhythmias after acidosis are dependent on CaMKII activation and are associated with an increase in SR Ca(2+) load, which appears to be mainly due to the increase in PT-PLN.

The aim of the present work conducted at the Refugio de Vida Silvestre Bahı´a Samborombo´n, Argentina, is to analyse the most relevant aspects of the life cycles of Chasmagnathus granulatus and Uca uruguayensis: their abundance, size distribution and sex ratio in order to be used for make management purposes. A total of 1200 individuals of U. uruguayensis (megalopae, juveniles and adults) and 957 individuals of C. granulatus (juveniles and adults) were collected

Spontaneously hypertensive rat (SHR) constitutes a genetic model widely used to study the natural evolution of hypertensive heart disease. Ca2+-handling alterations are known to occur in SHR. However, the putative modifications of Ca2+-handling proteins during the progression to heart failure (HF) are not well established. Moreover, the role of apoptosis in SHR is controversial. We investigated intracellular Ca2+, Ca2+-handling proteins and apoptosis in SHR vs. control Wistar rats (W) from 3 to 15 months (mo). Changes associated with the transition to HF (i.e. lung edema and decrease in midwall fractional shortening), occurred at 15 mo in 38% of SHR (SHRF). In SHRF, twitch and caffeine-induced Ca2+ transients, significantly decreased relative to 6/9 mo and 15 mo without HF signs. This decrease occurred in association with a decrease in the time constant of caffeine-Ca2+ transient decay and an increase in Na+/Ca2+ exchanger (NCX) abundance (p<0.05) with no changes in SERCA2a expression/activity. An increased Ca2+-calmodulin-kinase II activity, associated with an enhancement of apoptosis (TUNEL and Bax/Bcl2) was observed in SHR relative to W from 3 to 15 mo. Conclusions: 1. Apoptosis is an early and persistent event that may contribute to hypertrophic remodeling but would not participate in the contractile impairment of SHRF. 2. The increase in NCX expression/activity, associated with an increase in Ca2+ efflux from the cell, constitutes a primary alteration of Ca2+-handling proteins in the evolution to HF. 3. No changes in SERCA2a expression/activity are observed when HF signs become evident.

Previous results from our laboratory showed that phosphorylation of ryanodine receptor 2 (RyR2) by Ca(2+) calmodulin-dependent kinase II (CaMKII) was a critical but not the unique event responsible for the production of reperfusion-induced arrhythmogenesis, suggesting the existence of other mechanisms cooperating in an additive way to produce these rhythm alterations. Oxidative stress is a prominent feature of ischemia/reperfusion injury. Both CaMKII and RyR2 are proteins susceptible to alteration by redox modifications. This study was designed to elucidate whether CaMKII and RyR2 redox changes occur during reperfusion and whether these changes are involved in the genesis of arrhythmias. Langendorff-perfused hearts from rats or transgenic mice with genetic ablation of CaMKII phosphorylation site on RyR2 (S2814A) were subjected to ischemia-reperfusion in the presence or absence of a free radical scavenger (mercaptopropionylglycine, MPG) or inhibitors of NADPH oxidase and nitric oxide synthase. Left ventricular contractile parameters and monophasic action potentials were recorded. Oxidation and phosphorylation of CaMKII and RyR2 were assessed. Increased oxidation of CaMKII during reperfusion had no consequences on the level of RyR2 phosphorylation. Avoiding the reperfusion-induced thiol oxidation of RyR2 with MPG produced a reduction in the number of arrhythmias and did not modify the contractile recovery. Conversely, selective prevention of S-nitrosylation and S-glutathionylation of RyR2 was associated with higher numbers of arrhythmias and impaired contractility. In S2814A mice, treatment with MPG further reduced the incidence of arrhythmias. Taken together, the results suggest that redox modification of RyR2 synergistically with CaMKII phosphorylation modulates reperfusion arrhythmias.

The aim of the present work conducted at the Refugio de Vida Silvestre Bahía Samborombón is to analyse the most relevant aspects of the fecundity of Chasmagnathus granulatus and Uca uruguayensis. Samplings were carried out from March 2001 to February 2003. Ovigerous females of U. uruguayensis (N = 13) and C. granulatus (N = 25) were found during spring and summer, their sizes (CW) varied from 9.1 to 11.7 mm for the former species and from 22.8 to 32.4 mm for the latter. The egg diameter in U. uruguayensis ranged from 245 to 260 µm for embryos in the early stage of development and from 250 to 345 µm for those in mid-developmental stage, while in C. granulatus from 250 to 345 µm and from 260 to 365 µm respectively. Fecundity varied from 1126 to 6745 eggs/brood in U. uruguayensis and 15688-57418 eggs/brood in C. granulatus. For those females with broods in mid-developmental stage, several relationships were made. For U. uruguayensis the best correlation coefficients were obtained for the relationships: female weight vs. egg mass weight and carapace width vs. egg mass weight; for C. granulatus the best association was obtained between female size and the egg number and the egg mass weight.Keywords: fecundity, Uca uruguayensis, Chasmagnathus granulatus, Argentina. Fecundidade deUca uruguayensis e Chasmagnathus granulatus (Decapoda, Brachyura) no "Refúgio de Vida Silvestre" Bahía Samborombón, Argentina Resumo O objetivo do presente trabalho foi analisar os aspectos mais relevantes da fecundidade de Chasmagnathus granulatus e Uca uruguayensis no Refúgio da Vida Silvestre Bahia Samborombón. As amostragens foram realizadas de março de 2001 a fevereiro de 2003. As fêmeas ovígeras de U. uruguayensis (N = 13) e de C. granulatus (N = 25) foram capturadas na primavera e verão. A largura da carapaça (LC) de U. uruguayensis variou de 9.1 a 11.7 mm, e de 22.8 a 32.4 mm para C. granulatus. O diâmetro dos ovos de U. uruguayensis variou de 245 a 260 µm para embriões em estágio de desenvolvimento inicial e de 250 a 345 µm para aqueles em estágio intermediário; para C. granulatus as variações foram de 250 a 345 µm e de 260 a 365 µm, respectivamente. A fecundidade de U. uruguayensis variou de 1126 a 6745 ovos/desova e para C. granulatus de 15688 a 57418 ovos/desova. Para as fêmeas com massa de ovos em estágio de desenvolvimento intermediário foram efetuadas várias relações: para U. uruguayensis os melhores coeficientes de correlação foram obtidos nas relações: peso da fêmea vs. peso da massa de ovos, e largura da carapaça vs. peso da massa de ovos. Para C. granulatus, a melhor associação foi obtida entre o número de ovos e o peso da massa de ovos.

We investigated the biology and ecology of the tanaidacean Sinelobus stanfordi from the littoral water of Martín García Island at five sites along the coast chosen for their substrate characteristics, and classified the individuals according to size (mm) and ectosomatic characterinalysisstics as: manca III (0.60-1.19), juvenile male and female I (0.90-1.49), juvenile female II and copulatory male I (1.2-1.79), preparatory female I (1.8-2.39), preparatory female II (2.40-2.99), copulatory female I (2.40-2.69), copulatory female II (2.99-4.19), ovigerous female (1.80-3.59), copulatory male II (1.5-1.79), copulatory male III (2.10-2.69) and copulatory male IV (2.40-3.89). The smallest female with rudimentary ovisacs measured 1.84 mm. The tanaid density ranged from 0 to 10,548 ind.m-2. The average female-to-male sex ratio was 3.04:1. An abundance analysis indicated no significant differences among the sampling sites (X2 =4.037, p>0.001), while the fecundity (number of eggs) did not vary significantly with female size (r2 =0.2164, n = 19). The almost permanent presence of all developmental stages during every season of the year suggested a likely continuous year-long reproduction of S. stanfordi. Relationship between the water variables and S. stanfordi populations: The lower than optimal average electrical conductivity (160.24 μS.cm-1) for the species did not seem to limit the population-growth kinetics. The postmarsupial development throughout almost all the stages evidenced a significant positive correlation with pH. The soluble-reactive-phosphorus levels were within the low tolerance values as well as the NO-3 and the NH4 + and both close to optimal along with the near-optimal percent saturation of oxygen.

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