AtA6PR1 and AtA6PR2 encode putative aldose 6-phosphate reductases that are cytosolically localized and respond differentially to cold and salt stress in Arabidopsis thaliana

Sorbitol is synthesized in Rosaceae species, especially in source organs, in a pathway involving aldose 6-phosphate reductase (A6PR, EC 1.1.1.200). As compatible solutes, sorbitol and other sugar alcohols assist in the ability of the plant to withstand abiotic stress conditions. Here, we identify two tandemly-duplicated genes in a non-Rosaceae species (Arabidopsis thaliana L.), and show that the proteins encoded by At2g21250 (AtA6PR1) and At2g21260 (AtA6PR2) possess the molecular characteristics of A6PRs. Consistent with bioinformatic predictions, we determined that green fluorescent protein-tagged versions of AtA6PR1 and AtA6PR2 are cytosolically localized, a finding supported by immunoblotting using a specific anti-AtA6PR1 antisera after subcellular fractionation of Arabidopsis leaves. We also show that under standard growth conditions, both genes are widely-expressed, whilst AtA6PR1 protein accumulates in both source and sink organs. Under short term (24 h) cold (4 °C) or saline (150 mM NaCl) stress, both genes respond differentially and reciprocally in leaf and root tissues, suggesting a sub-functionalization of their roles. The identification of homozygous T-DNA insertional ata6pr1-mutants will facilitate the functional characterization of this gene, and help to determine its role under abiotic stress conditions.

     

Deciphering the principles that govern mutually exclusive expression of Plasmodium falciparum clag3 genes

The product of the Plasmodium falciparum genes clag3.1 and clag3.2 plays a fundamental role in malaria parasite biology by determining solute transport into infected erythrocytes. Expression of the two clag3 genes is mutually exclusive, such that a single parasite expresses only one of the two genes at a time. Here we investigated the properties and mechanisms of clag3 mutual exclusion using transgenic parasite lines with extra copies of clag3 promoters located either in stable episomes or integrated in the parasite genome. We found that the additional clag3 promoters in these transgenic lines are silenced by default, but under strong selective pressure parasites with more than one clag3 promoter simultaneously active are observed, demonstrating that clag3 mutual exclusion is strongly favored but it is not strict. We show that silencing of clag3 genes is associated with the repressive histone mark H3K9me3 even in parasites with unusual clag3 expression patterns, and we provide direct evidence for heterochromatin spreading in P. falciparum. We also found that expression of a neighbor ncRNA correlates with clag3.1 expression. Altogether, our results reveal a scenario where fitness costs and non-deterministic molecular processes that favor mutual exclusion shape the expression patterns of this important gene family.     

Extracellular proteinases from the phytopathogenic fungus Fusarium culmorum

The growth of Fusarium culmorum fungus on a medium containing thermostable proteins from potato tubers was accompanied by the production of proteinases, exhibiting activity over a broad pH range (from 6.0–10.0). When studied by SDS-PAGE in the presence of β-mercaptoethanol, extracellular proteinases were represented by at least five species with a molecular weight of 30–60 kDa. Inhibitor analysis and studies of enzyme activities with synthetic substrates demonstrated that the culture liquid of Fusarium culmorum contained serine proteinases of various classes. The amount of subtilisin-like proteinases was the highest. A near-complete inhibition of the enzymes was caused by proteinaceous proteinase inhibitors from potato tubers. These data suggest that proteinases of the phytopathogen Fusarium culmorum serve as a metabolic target for natural inhibitors of potato proteinases.     

Feedback on the Rate and Depth of Chest Compressions during Cardiopulmonary Resuscitation Using Only Accelerometers

Background

Quality of cardiopulmonary resuscitation (CPR) is key to increase survival from cardiac arrest. Providing chest compressions with adequate rate and depth is difficult even for well-trained rescuers. The use of real-time feedback devices is intended to contribute to enhance chest compression quality. These devices are typically based on the double integration of the acceleration to obtain the chest displacement during compressions. The integration process is inherently unstable and leads to important errors unless boundary conditions are applied for each compression cycle. Commercial solutions use additional reference signals to establish these conditions, requiring additional sensors. Our aim was to study the accuracy of three methods based solely on the acceleration signal to provide feedback on the compression rate and depth.

Materials and Methods

We simulated a CPR scenario with several volunteers grouped in couples providing chest compressions on a resuscitation manikin. Different target rates (80, 100, 120, and 140 compressions per minute) and a target depth of at least 50 mm were indicated. The manikin was equipped with a displacement sensor. The accelerometer was placed between the rescuer’s hands and the manikin’s chest. We designed three alternatives to direct integration based on different principles (linear filtering, analysis of velocity, and spectral analysis of acceleration). We evaluated their accuracy by comparing the estimated depth and rate with the values obtained from the reference displacement sensor.

Results

The median (IQR) percent error was 5.9% (2.8–10.3), 6.3% (2.9–11.3), and 2.5% (1.2–4.4) for depth and 1.7% (0.0–2.3), 0.0% (0.0–2.0), and 0.9% (0.4–1.6) for rate, respectively. Depth accuracy depended on the target rate (p < 0.001) and on the rescuer couple (p < 0.001) within each method.

Conclusions

Accurate feedback on chest compression depth and rate during CPR is possible using exclusively the chest acceleration signal. The algorithm based on spectral analysis showed the best performance. Despite these encouraging results, further research should be conducted to asses the performance of these algorithms with clinical data.     

Probiotic Lactobacilli Administration Induces Changes in the Fecal Microbiota of Preweaned Dairy Calves

Probiotics and Antimicrobial Proteins - Early microbial colonization is a determinant factor in animal health, and probiotic administration has been demonstrated to modulate intestinal microbiota...     

Leaf herbivory and fluctuating asymmetry as indicators of mangrove stress

Wetlands Ecology and Management - Fluctuating asymmetry (FA), a widely used measure of developmental instability in plants and animals, which describes random differences in size and/or shape...     

TNF triggers mitogenic signals in NIH 3T3 cells but induces apoptosis when the cell cycle is blocked

Tumor necrosis factor (TNF) is known to be a mediator of a variety of cellular responses including apoptotic death or proliferation depending on the target cell and the environmental conditions. We show here that TNF triggers both growth and death signals in NIH 3T3 murine fibroblasts. In cells arrested in G(0) by serum deprivation, TNF drives approximately 50% of them to enter the cell cycle, but kills the cells that remain quiescent. The presence of serum prevents toxic effects of TNF, suggesting that TNF can cooperate to drive cells through the cell cycle, but is unable to do so by itself and alternatively it triggers death signals in cells unable to proliferate. Interestingly, TNF induces a similar toxic effect in cells forced to stay at the G(1)/S border, S or M phases. We have explored the TNF apoptotic pathway in arrested cells. This mechanism is not due to the loss of the anti-apoptotic capacity of NFkappaB and is mediated by mitochondria since Bcl-2 overexpression partially inhibits cell death. There are, however, interesting differences in the kinetics of mitochondrial events which indicate that this form of sensitization to TNF leads to an apoptotic mechanism different from that observed after sensitization by RNA synthesis inhibition.