Regulation of durum wheat Na<Superscript>+</Superscript>/H<Superscript>+</Superscript> exchanger TdSOS1 by phosphorylation

We have identified a plasma membrane Na⁺/H⁺ exchanger from durum wheat, designated TdSOS1. Heterologous expression of TdSOS1 in a yeast strain lacking endogenous Na⁺ efflux proteins showed complementation of the Na⁺- and Li⁺-sensitive phenotype by a mechanism involving cation efflux. Salt tolerance conferred by TdSOS1 was maximal when co-expressed with the Arabidopsis protein kinase complex SOS2/SOS3. In vitro phosphorylation of TdSOS1 with a hyperactive form of the Arabidopsis SOS2 kinase (T/DSOS2∆308) showed the importance of two essential serine residues at the C-terminal hydrophilic tail (S1126, S1128). Mutation of these two serine residues to alanine decreased the phosphorylation of TdSOS1 by T/DSOS2∆308 and prevented the activation of TdSOS1. In addition, deletion of the C-terminal domain of TdSOS1 encompassing serine residues at position 1126 and 1128 generated a hyperactive form that had maximal sodium exclusion activity independent from the regulatory SOS2/SOS3 complex. These results are consistent with the presence of an auto-inhibitory domain at the C-terminus of TdSOS1 that mediates the activation of TdSOS1 by the protein kinase SOS2. Expression of TdSOS1 mRNA in young seedlings of the durum wheat variety Om Rabia3, using different abiotic stresses (ionic and oxidative stress) at different times of exposure, was monitored by RT–PCR.     

Problemas de seguridad relacionados con la aplicación de dispositivos de restricción física en personas mayores

The use of physical restraint devices on frail elderly could have significant negative consequences on their health. Apart from complications due to prolonged immobility, the use of this procedure is associated with other serious adverse effects which occur when a person is restricted in a position which carries a risk of asphyxiation. The devices most implicated in these incidents are bedrails, vests and restraining belts. Physical restraint could also be associated as much with the sudden death of patients, due to the stress that it causes, as with injuries from falling. This article presents the recommendations which experts, manufacturers and institutions dealing with the quality of health care have issued for the safest use of this procedure. It stresses the need for better training of professionals, as well as the importance of investigating the factors which can lead to accidents with the aim of preventing them.     

Safety problems associated with using physical restraint devices on the elderly

The use of physical restraint devices on frail elderly could have significant negative consequences on their health. Apart from complications due to prolonged immobility, the use of this procedure is associated with other serious adverse effects which occur when a person is restricted in a position which carries a risk of asphyxiation. The devices most implicated in these incidents are bedrails, vests and restraining belts. Physical restraint could also be associated as much with the sudden death of patients, due to the stress that it causes, as with injuries from falling. This article presents the recommendations which experts, manufacturers and institutions dealing with the quality of health care have issued for the safest use of this procedure. It stresses the need for better training of professionals, as well as the importance of investigating the factors which can lead to accidents with the aim of preventing them.     

Sustained release of PI3K inhibitor from PHA nanoparticles and in vitro growth inhibition of cancer cell lines

The phosphoinositide-3-kinases (PI3Ks) are a conserved family of lipid kinases that phosphorylate the 3-hydroxyl group of phosphatidylinositols in response to extracellular stimuli. PI3K pathway is enrolled in different kinds of human cancer and plays a prominent role in cancer cell growth and survival. Several PI3K inhibitors have been recently identified but some PI3K inhibitors with high potency in vitro do not show satisfactory effects in animal cancer models because of the poor pharmaceutical properties in vivo such as poor solubility, instability, and fast plasma clearance rate. In this study, we developed a sustained release system of PI3K inhibitor (TGX221) based on polyhydroxyalkanoate nanoparticles (NP) and used it to block proliferation of cancer cell lines. TGX221 was gradually released from PHA-based NP and growth of cancer cell lines was significantly slower in NP-TGX221-treated cells than in either negative controls or in cells receiving free TGX221. Since poor bioavailability and limited in vivo half-life are common features of hydrophobic PI3K inhibitors, our results open the way to similar formulation of other PI3K blockers and to new strategies in cancer treatment.     

Inhibition of mitochondrial respiratory chain in the brain of rats after hepatic failure induced by acetaminophen

To explore the effect of LYRM1 over-expression on basal and insulin-stimulated glucose uptake in rat skeletal muscle cells, and to understand the underlying mechanisms, Rat myoblasts (L6) transfected with either an empty expression vector (pcDNA3.1Myc/His B) or a LYRM1 expression vector were differentiated into myotubes. Glucose uptake was determined by measuring 2-deoxy-D-[(3)H] glucose uptake into L6 myotubes. Western blotting was performed to assess the translocation of insulin-sensitive glucose transporter 4 (GLUT4). It was also used to measure the phosphorylation and total protein contents of insulin-signaling proteins, such as the insulin receptor (IR), insulin receptor substrate (IRS)-1, phosphatidylinositol-3-kinase (PI3K) p85, Akt, ERK1/2, P38, and JNK. LYRM1 over-expression in L6 myotubes reduced insulin-stimulated glucose uptake and impaired insulin-stimulated GLUT4 translocation. It also diminished insulin-stimulated tyrosine phosphorylation of IRS-1, PI3K (p85), and serine phosphorylation of Akt without affecting the phosphorylation of IR, ERK1/2, P38, and JNK. LYRM1 regulates the function of IRS-1, PI3K, and Akt, and decreases GLUT4 translocation and glucose uptake in response to insulin. These observations highlight the potential role of LYRM1 in glucose homeostasis and possibly in the pathophysiology of type 2 diabetes related to obesity.     

Validation of the educational potential of a simulator to develop abilities and skills for the creation and maintenance of an intravenous cannula

Historically, the method used in veterinary medicine to teach technical procedures which are to be performed on live animals, has taken the form of an apprenticeship. However, in the last decade, there have been several new developments in technologies oriented toward the development of such abilities and skills by students - for example, manuals, videos, pictures, and virtual reality simulators. Unfortunately, these simulators are inaccessible to many, due to their high cost. For this reason, it is necessary to create simulators that are easy to manufacture at low cost, and that are also portable. These simulators also need to be validated with regard to their ability to fulfil the required educational objectives. The validation of a venous simulator is described in this study. Fifty-two veterinary students, with no previous experience in the creation and maintenance of an indwelling venous cannula, were selected at random. They were divided into two groups: one experimental group (n = 35), who had training practice on the simulator, and the remainder (n = 17), who acted as the control group (i.e. they had no training practice on the simulator). The outcome measure was the number of attempts required to successfully cannulate the cephalic vein of an anaesthetised rabbit. The students in the experimental group showed more skill in cannulating the vein, with 45% effectiveness, as compared to 20% effectiveness in the control group. The difference between the groups was statistically significant (p < 0.05).     

Alterations in Inflammatory Mediators, Oxidative Stress Parameters and Energetic Metabolism in the Brain of Sepsis Survivor Rats

Sepsis is characterized by biochemical alterations in the central nervous system at early times and cognitive impairment at late times after induction in sepsis animal model. In order to understand at least in part the mechanism of disease, we have evaluated the effects of sepsis on cytokine levels in the cerebrospinal fluid (CSF); oxidative parameters; the activity of the electron transport chain enzymes; and creatine kinase (CK) activity in the brain of sepsis survivor rats 10 days after cecal ligation and perforation (CLP). Male Wistar rats underwent CLP with "basic support" or sham-operated. Ten days after surgery, the animals were killed and prefrontal cortex, cortex, hippocampus, striatum, cerebellum, and CSF were obtained. It was found a decrease in the levels of TNF-α (P = 0.001), IL-1β (P = 0.008), IL-6 (P = 0.038), and IL-10 (P = 0.022) in the CSF; an increase in the TBARS only hippocampus (0.027); an up-regulation in the activity of complex II (P = 0.024), III (P = 0.018), and IV (P = 0.047) only in the prefrontal cortex; a decrease in the CK activity in the cerebellum (P = 0.001) and striatum (P = 0.0001), and an increase in the hippocampus (P = 0.0001) and cortex (P = 0.0001). Oxidative stress and mitochondrial alterations observed during early times in sepsis, persisted up to 10 days after surgery. The cytokines levels during the early times were found at high levels, decreasing to low levels after 10 days. In conclusion, these findings may contribute for a better comprehension of the cognitive damage in sepsis survivor rats.     

QualiTree, a virtual fruit tree to study the management of fruit quality. II. Parameterisation for peach, analysis of growth-related processes and agronomic scenarios

In this paper, QualiTree, a fruit tree model designed to study the management of fruit quality, and developed and described in a companion paper (Lescourret et al. in Trees Struct Funct, 2010), was combined with a simple light-interception sub-model, and then parameterised and tested on peach in different situations. Simulation outputs displayed fairly good agreement with the observed data concerning mean fruit and vegetative growth. The variability over time of fruit and vegetative growth was well predicted. QualiTree was able to reproduce the observed response of trees to heterogeneous thinning treatments in terms of fruit growth. A sensitivity analysis showed that the average seasonal growth rates of the different organs were sensitive to changes to the values of their respective initial relative growth rates and that stem wood was the tree organ the most affected by a change in the initial relative growth rates of other organs. QualiTree was able to react to simulated scenarios that combined thinning and pest attacks. As expected, thinning intensity and the percentage damage caused by pests significantly affected fruit yield and quality traits at harvest. These simulations showed that QualiTree could be a useful tool to design innovative horticultural practices.