Effect of physical training on the adipose tissue of diet-induced obesity mice: interaction between reactive oxygen species and lipolysis.

It is well known that high-fat diets (HFDs) induce obesity and result in an increase in oxidative stress in adipose tissue, which leads to an impairment of fat mobilization by a downregulation of the lipases, such as hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL). On the other hand, exercise training leads to a reduction in adipose tissue and an improvement of antioxidant status and the lipolytic pathway. Our aim was to examine the influence of exercise and moderate intensity training on oxidative stress parameters and the relationship between the proteins involved in the lipolysis of animals subjected to a high-fat fed diet. Twenty-four mice were used and divided into 4 groups (n=6): standard diet (SD); standard diet plus exercise (SD+Ex); high-fat diet (HFD); and high-fat diet plus exercise (HFD+Ex). The animals received HFD for 90 days and submitted to a daily training protocol in swinging. The animals were euthanized 48 h after the last session of exercise. White adipose tissue epididymal fat was excised for the measurement of oxidative stress parameters and protein levels of lipolytic enzymes by Western blotting. The results show an increase in body weight after 90 days of HFD, and exercise training prevented great gain. In adipose tissue, lipid peroxidation and protein carbonylation increased after HFD and decreased significantly after exercise training. The protein level of CGI-58 was reduced, and FAS was increased in the HFD than in SD, whereas ATGL exhibited an increase (p<0.05) in HFD than in SD. The exercise plays a significant role in reducing oxidative damage, along with the regulation of proteins that are involved in the lipolysis of animals exposed to HFD.

Exercise training performed simultaneously to a high-fat diet reduces the degree of insulin resistance and improves adipoR1-2/APPL1 protein levels in mice.

BACKGROUND: The aim of the present study was to evaluate the protective effect of concurrent exercise in the degree of the insulin resistance in mice fed with a high-fat diet, and assess adiponectin receptors (ADIPOR1 and ADIPOR2) and endosomal adaptor protein APPL1 in different tissues. METHODS: Twenty-four mice were randomized into four groups (n = 6): chow standard diet and sedentary (C); chow standard diet and simultaneous exercise training (C-T); fed on a high-fat diet and sedentary (DIO); and fed on a high-fat diet and simultaneous exercise training (DIO-T). Simultaneously to starting high-fat diet feeding, the mice were submitted to a swimming exercise training protocol (2 x 30 minutes, with 5 minutes of interval/day), five days per week, for twelve weeks (90 days). Animals were then euthanized 48 hours after the last exercise training session, and adipose, liver, and skeletal muscle tissue were extracted for an immunoblotting analysis. RESULTS: IR, IRs, and Akt phosphorylation decreased in the DIO group in the three analyzed tissues. In addition, the DIO group exhibited ADIPOR1 (skeletal muscle and adipose tissue), ADIPOR2 (liver), and APPL1 reduced when compared with the C group. However, it was reverted when exercise training was simultaneously performed. In parallel, ADIPOR1 and 2 and APPL1 protein levels significantly increase in exercised mice. CONCLUSIONS: Our findings demonstrate that exercise training performed concomitantly to a high-fat diet reduces the degree of insulin resistance and improves adipoR1-2/APPL1 protein levels in the hepatic, adipose, and skeletal muscle tissue.

Phase IV study comparing diurnal glycemic profile following the administration of 2 NPH plus regular human DNA recombinant insulin regimens in type 1 diabetes mellitus (T1DM) adult patients.

Intensive insulin therapy (IIT) based on multiple daily injections of long plus rapid-acting insulin has been demonstrated to reduce mortality and morbidity associated with chronic hyperglycemia in T1DM patients. The objective of this study was to assess and compare the postprandial glycemic profile over a diurnal 12 h-period produced by the administration of a new NPH plus regular human DNA recombinant IIT (test regimen) relative to the reference IIT in T1DM patients. A phase IV, single-center, open-label, randomized, multiple-dose, balanced, cross-over study in 12 T1DM patients was conducted. Patients were assigned to receive either the test (Densulin® N (NPH) plus Densulin® R (regular),100 UI/ml, Denver Farma, Argentina) followed by the reference (InsulatardHM® (NPH) plus ActrapidHM®,100 UI/ml, Novo Nordisk Pharma Argentina) regimens or viceversa, according to a random sequence. Each treatment regimen consisted of 2 phases of an ambulatory run-in period of 7 days followed by 12 h confinement period. Blood glucose levels were measured. Glycemic profile was evaluated through glycemic plasma-concentration time curves, area under the time-concentration glycemic curves from basal to 2 h (GlyAUC0-2) and to 12 h (GlyAUC0-12) postprandial, and maximum glycemic postprandial concentration (GlyCmax). 12 hour glycemic concentration-time curves were similar for both test and reference regimens. Geometric least square means ratios Test/ref regimens and their 90% confidence interval for GlyAUC0-2, GlyAUC0-12 and GlyCmax were 94.33 (81.13-125.09), 107.75 (94.05-123.45) and 105 (92.89-118.68), respectively. Both regimens presented similar safety profile. This study demonstrated that the new human DNA recombinant NPH and regular insulin is equally effective to the reference regimen for postprandial diurnal glycemic profile.

Effects of dietary Spirulina on vascular reactivity.

There are several reports suggesting that Spirulina (Arthrospira) may have a beneficial effect in the prevention of cardiovascular diseases. Here we review the results of studies on the effects of dietary Spirulina on the vasomotor reactivity of aortic rings excised from either lean or obese Wistar rats. We also review preliminary results on the effects of Spirulina intake on plasma lipids and blood pressure in humans. The results of the former studies strongly suggest that Spirulina induces a tone-related increase in the synthesis/release of nitric oxide by the endothelium as well as an increase in the synthesis/release of a vasodilating cyclooxygenase-dependent metabolite of arachidonic acid and/or a decrease in the synthesis/release of a vasoconstricting eicosanoid by the endothelium. In humans, Spirulina maxima intake decreases blood pressure and plasma lipid concentrations, especially triacylglycerols and low-density lipoprotein-cholesterol, and indirectly modifies the total cholesterol and high-density lipoprotein-cholesterol values.

Oviposition and predation by Speciomerus revoili (Coleoptera, Bruchidae) on seeds of Acrocomia aculeata (Arecaceae) in Brasília, DF, Brazil.

Oviposition and predation levels by Speciomerus revoili bruchid beetles were quantified on fruits and seeds of the macaúba palm, Acrocomia aculeata, collected from below mother-trees within the Sarah Kubitschek Park of Brasília, DF, Brazil. A maximum of 12 eggs per fruit were found, with high variations observed between samples. No clear pattern was found for the distribution of the number of eggs per fruit, perhaps due to the artificial conditions of the study area, the absence of dispersers and/or the plasticity in the oviposition behavior of the insect. The number of eggs per fruit was not related to fruit size, but was associated with their availability under the tree-mother. This suggests that the density of eggs per fruit is a balance between the availability of this resource and the number of females in the beetle population. The observed mortality rate, from the egg phase to the final larval stages, was over 75%. About 40% of the seeds of Acrocomia aculeata were predated by Speciomerus revoili.

Long-term depolarization regulates the alpha1s, subunit of skeletal muscle Ca2+ channels and the amplitude of L-type Ca2+ currents.

The effects of long-term depolarization on the level of alpha1s and on L-type Ca2+ currents of skeletal muscle were investigated. Long-term depolarization (14 h) caused a 50% decrease of alpha1s, revealed with the Western blot technique. This decline was prevented by preincubation with the Ca2+ channel blocker nifedipine. Electrophysiological experiments using the voltage-clamp technique were performed to measure the actions of long-term depolarization on Ca2+ currents and charge movement. A progressive decline in the amplitude of the Ca2+ currents by depolarizations lasting 0.5-14 h was observed. Similar to Western blot results, the fall in current amplitude was prevented by nifedipine, and it depended on external Ca2+. The nonlinear charge mobilized by step pulses was also significantly reduced (50%) by long-term depolarization. It is suggested that alpha1s subunit is down-regulated by long-term depolarization by a very stringent mechanism and that, in this process, Ca2+ ions permeating through L-type channels play a key role. A new role for the L-type Ca2+ current in skeletal muscle fibers in which the channels are self-regulated is proposed.

Oviposition and predation by Speciomerus revoili (Coleoptera, Bruchidae) on seeds of Acrocomia aculeata (Arecaceae) in Brasília, DF, Brazil

Oviposition and predation levels by Speciomerus revoili bruchid beetles were quantified on fruits and seeds of the macaúba palm, Acrocomia aculeata, collected from below mother-trees within the Sarah Kubitschek Park of Brasília, DF, Brazil. A maximum of 12 eggs per fruit were found, with high variations observed between samples. No clear pattern was found for the distribution of the number of eggs per fruit, perhaps due to the artificial conditions of the study area, the absence of dispersers and/or the plasticity in the oviposition behavior of the insect. The number of eggs per fruit was not related to fruit size, but was associated with their availability under the tree-mother. This suggests that the density of eggs per fruit is a balance between the availability of this resource and the number of females in the beetle population. The observed mortality rate, from the egg phase to the final larval stages, was over 75 percent. About 40 percent of the seeds of Acrocomia aculeata were predated by Speciomerus revoili

Functional properties of a truncated recombinant GIRK5 potassium channel.

Xenopus laevis oocytes codify a G-protein-activated inward rectifier potassium channel (GIRK5 or Kir3.5). Coinjection of other GIRKs, the muscarinic m2 receptor, or Gbetagamma protein cRNAs is required to observe functional GIRKx-GIRK5 heteromultimers in oocytes. Studies with GIRK2 isoforms have shown that the size of the amino or carboxyl terminus plays a crucial role on giving functional K(+) channels. In this work we studied the properties of a GIRK5 with 25 amino acids deleted toward its amino-terminal domain. Injection of GIRK5-Delta25 cRNA alone displayed large basal and transient inward rectifying currents in oocytes. The instantaneous currents reached a stationary level after a long duration voltage pulse (10 s). For this relaxation, fast (tau(1)) and slow (tau(2)) time constants were estimated at different voltages. Recovery from inactivation followed a monoexponential function (tau=0.95+/-0.07 s). By contrast with other inward rectifier channels, blockade of GIRK5-Delta25 by extracellular Ba(2+) was voltage-independent (K(d)=102+/-2 microM), suggesting the presence of a Ba(2+) site at the external channel vestibule. To confirm this hypothesis, the Ba(2+) sensitivity of two charged mutants GIRK5-Delta25(N129E) and GIRK5-Delta25(K157E) at each of the external loops was determined. GIRK5-Delta25(N129E) and GIRK5-Delta25(K157E) showed a 100-fold and 2-fold higher affinity to Ba(2+), respectively, supporting the existence of this Ba(2+) binding site.

A long-term blockade of L-type calcium currents upregulates the number of Ca2+ channels in skeletal muscle.

The effects of a long-term blockade of L-type Ca2+ channels on membrane currents and on the number of dihydropyridine binding sites were investigated in skeletal muscle fibers. Ca2+ currents (ICa) and intramembrane charge movement were monitored using a voltage-clamp technique. The peak amplitude of ICa increased by more than 40% in fibers that were previously incubated for 24 hr in solutions containing the organic Ca2+ channel blocker nifedipine or in Ca2+-free conditions. A similar incubation period with Cd2+, an inorganic blocker, produced a moderate increase of 20% in peak ICa. The maximum mobilized charge (Qmax) increased by 50% in fibers preincubated in Ca2+-free solutions or in the presence of Cd2+. Microsomal preparations from frog skeletal muscle were isolated by differential centrifugation. Preincubation with Cd2+ prior to the isolation of the microsomal fraction doubled the number of 3H-PN200-110 binding sites and produced a similar increase in the values of the dissociation constant. The increase in the number of binding sites is consistent with the increase in the peak amplitude of ICa as well as with the increase in Qmax.

Modulation of a calcium-activated chloride current by Maitotoxin.

The effect of Maitotoxin (MTX) on the calcium-activated chloride current (ICl-Ca) from Xenopus oocytes was studied, applying the two-electrode voltage clamp technique. MTX increased the current amplitude at all the voltages explored and reduced the time to reach the maximum current level (time to peak). At low toxin concentrations (15 pM), both effects were fully reversible. Activation of ICl-Ca by MTX was secondary to the increment in the intracellular Ca2+ concentration induced by this toxin, since incubation of the oocytes with the cell-permeant Ca2+ chelator BAPTA-AM, greatly reduced the effect of MTX on ICl-Ca. Furthermore, external chloride ions removal also diminished the MTX effect on the current, strongly suggesting that the main current activated by MTX is ICl-Ca. Subsequent applications of a fixed toxin concentration after toxin washout resulted in enhanced ICl-Ca, suggesting that the toxin effect potentiates.

Modulation of Ca2+ channels, charge movement and Ca2+ transients by heparin in frog skeletal muscle fibres.

This study is an investigation into the modulatory effects of heparin, a component of the extracellular matrix that binds to dihydropyridine receptors, on contraction and Ca2+ channels in frog skeletal muscle. Using tension and Ca2+ signal measurements in single intact skeletal muscle cells we have found that heparin (100-200 micrograms ml-1) substantially potentiates twitch and tetanic tension (55% and 28%, respectively). In contrast, heparin reduces the amplitude of K+ contractures. Heparin most likely potentiates twitch tension by prolonging action potentials. The ionic basis of this effect was investigated in voltage-clamp experiments. Membrane currents were monitored in voltage-clamped segments of single fibres using the triple Vaseline gap technique. We found that heparin partially blocks delayed rectifier potassium channels. The depressive effects of heparin on K+ contractures prompted us to investigate the effects of heparin on charge movement and Ca2+ currents (ICa) under voltage-clamp. Charge movement was measured using a subtraction procedure that employed a -20 mV control pulse from a holding potential of -100 mV. Heparin depresses the total charge by 25%. We propose that the reduction in the amplitude of potassium contractures is related to a partial blockade of charge movement. Extracellular heparin shifts the ICa-V relation toward more negative voltages and delays the deactivation of tail currents. Double pulse experiments revealed that conditioning depolarizations speed the activation of ICa during test depolarizations. Heparin does not affect this process. The primary action of heparin is to accelerate the activation of ICa during pulses not preceded by conditioning depolarizations. Overall, the kinetic effects of heparin on ICa would increase the Ca2+ influx associated with action potentials. However, mechanical and optical experiments performed in Ca(2+) -free solutions and in the presence of Ca2+ channel blockers revealed that twitch and tetanic potentiation occur even in the absence of Ca(2+) -influx.