Telomere length regulation: coupling DNA end processing to feedback regulation of telomerase

The conventional DNA polymerase machinery is unable to fully replicate the ends of linear chromosomes. To surmount this problem, nearly all eukaryotes use the telomerase enzyme, a specialized reverse transcriptase that utizes its own RNA template to add short TG-rich repeats to chromosome ends, thus reversing their gradual erosion occurring at each round of replication. This unique, non-DNA templated mode of telomere replication requires a regulatory mechanism to ensure that telomerase acts at telomeres whose TG tracts are too short, but not at those with long tracts, thus maintaining the protective TG repeat cap at an appropriate average length. The prevailing notion in the field is that telomere length regulation is brought about through a negative feedback mechanism that counts TG repeat-bound protein complexes to generate a signal that regulates telomerase action. This review summarizes experiments leading up to this model and then focuses on more recent experiments, primarily from yeast, that begin to suggest how this counting mechanism might work. The emerging picture is that of a complex interplay between the conventional DNA replication machinery, DNA damage response factors, and a specialized set of proteins that help to recruit and regulate the telomerase enzyme

Plasticity of inhibition.

Until recently, the study of plasticity of neural circuits focused almost exclusively on potentiation and depression at excitatory synapses on principal cells. Other elements in the neural circuitry, such as inhibitory synapses on principal cells and the synapses recruiting interneurons, were assumed to be relatively inflexible, as befits a role of inhibition in maintaining stable levels and accurate timing of neuronal activity. It is now evident that inhibition is highly plastic, with multiple underlying cellular mechanisms. This Review considers these recent developments, focusing mainly on functional and structural changes in GABAergic inhibition of principal cells and long-term plasticity of glutamateric recruitment of inhibitory interneurons in the mammalian forebrain. A major challenge is to identify the adaptive roles of these different forms of plasticity, taking into account the roles of inhibition in the regulation of excitability, generation of population oscillations, and precise timing of neuronal firing

An approach to holistically assess (dairy) farm eco-efficiency by combining Life Cycle Analysis with Data Envelopment Analysis models and methodologies

Eco-efficiency is a useful guide to dairy farm sustainability analysis aimed at increasing output (physical or value added) and minimizing environmental impacts (EIs). Widely used partial eco-efficiency ratios (EIs per some functional unit, e.g. kg milk) can be problematic because (i) substitution possibilities between EIs are ignored, (ii) multiple ratios can complicate decision making and (iii) EIs are not usually associated with just the functional unit in the ratio's denominator. The objective of this study was to demonstrate a 'global' eco-efficiency modelling framework dealing with issues (i) to (iii) by combining Life Cycle Analysis (LCA) data and the multiple-input, multiple-output production efficiency method Data Envelopment Analysis (DEA). With DEA each dairy farm's outputs and LCA-derived EIs are aggregated into a single, relative, bounded, dimensionless eco-efficiency score, thus overcoming issues (i) to (iii). A novelty of this study is that a model providing a number of additional desirable properties was employed, known as the Range Adjusted Measure (RAM) of inefficiency. These properties altogether make RAM advantageous over other DEA models and are as follows. First, RAM is able to simultaneously minimize EIs and maximize outputs. Second, it indicates which EIs and/or outputs contribute the most to a farm's eco-inefficiency. Third it can be used to rank farms in terms of eco-efficiency scores. Thus, non-parametric rank tests can be employed to test for significant differences in terms of eco-efficiency score ranks between different farm groups. An additional DEA methodology was employed to 'correct' the farms' eco-efficiency scores for inefficiencies attributed to managerial factors. By removing managerial inefficiencies it was possible to detect differences in eco-efficiency between farms solely attributed to uncontrollable factors such as region. Such analysis is lacking in previous dairy studies combining LCA with DEA. RAM and the 'corrective' methodology were demonstrated with LCA data from French specialized dairy farms grouped by region (West France, Continental France) and feeding strategy (regardless of region). Mean eco-efficiency score ranks were significantly higher for farms with 30% maize in the total forage area before correcting for managerial inefficiencies. Mean eco-efficiency score ranks were higher for West than Continental farms, but significantly higher only after correcting for managerial inefficiencies. These results helped identify the eco-efficiency potential of each region and feeding strategy and could therefore aid advisors and policy makers at farm or region/sector level. The proposed framework helped better measure and understand (dairy) farm eco-efficiency, both within and between different farm groups

The relationship of dairy farm eco-efficiency with intensification and self-sufficiency. Evidence from the French dairy sector using Life Cycle Analysis, Data Envelopment Analysis and Partial Least Squares Structural Equation Modelling

We aimed at quantifying the extent to which agricultural management practices linked to animal production and land use affect environmental outcomes at a larger scale. Two practices closely linked to farm environmental performance at a larger scale are farming intensity, often resulting in greater off-farm environmental impacts (land, non-renewable energy use etc.) associated with the production of imported inputs (e.g. concentrates, fertilizer); and the degree of self-sufficiency, i.e. the farm’s capacity to produce goods from its own resources, with higher control over nutrient recycling and thus minimization of losses to the environment, often resulting in greater on-farm impacts (eutrophication, acidification etc.). We explored the relationship of these practices with farm environmental performance for 185 French specialized dairy farms. We used Partial Least Squares Structural Equation Modelling to build, and relate, latent variables of environmental performance, intensification and self-sufficiency. Proxy indicators reflected the latent variables for intensification (milk yield/cow, use of maize silage etc.) and self-sufficiency (home-grown feed/total feed use, on-farm energy/total energy use etc.). Environmental performance was represented by an aggregate ‘eco-efficiency’ score per farm derived from a Data Envelopment Analysis model fed with LCA and farm output data. The dataset was split into two spatially heterogeneous (bio-physical conditions, production patterns) regions. For both regions, eco-efficiency was significantly negatively related with milk yield/cow and the use of maize silage and imported concentrates. However, these results might not necessarily hold for intensive yet more self-sufficient farms. This requires further investigation with latent variables for intensification and self-sufficiency that do not largely overlap- a modelling challenge that occurred here. We conclude that the environmental ‘sustainability’ of intensive dairy farming depends on particular farming systems and circumstances, although we note that more self-sufficient farms may be preferable when they may benefit from relatively low land prices and agri-environment schemes aimed at maintaining grasslands

Pleosporales

One hundred and five generic types of Pleosporales are described and illustrated. A brief introduction and detailed history with short notes on morphology, molecular phylogeny as well as a general conclusion of each genus are provided. For those genera where the type or a representative specimen is unavailable, a brief note is given. Altogether 174 genera of Pleosporales are treated. Phaeotrichaceae as well as Kriegeriella, Zeuctomorpha and Muroia are excluded from Pleosporales. Based on the multigene phylogenetic analysis, the suborder Massarineae is emended to accommodate five families, viz. Lentitheciaceae, Massarinaceae, Montagnulaceae, Morosphaeriaceae and Trematosphaeriaceae

Involvement of Iron in Biofilm Formation by Staphylococcus aureus

Staphylococcus aureus is a human pathogen that forms biofilm on catheters and medical implants. The authors' earlier study established that 1,2,3,4,6-penta-O-galloyl-β-D-glucopyranose (PGG) inhibits biofilm formation by S. aureus by preventing the initial attachment of the cells to a solid surface and reducing the production of polysaccharide intercellular adhesin (PIA). Our cDNA microarray and MALDI-TOF mass spectrometric studies demonstrate that PGG treatment causes the expression of genes and proteins that are normally expressed under iron-limiting conditions. A chemical assay using ferrozine verifies that PGG is a strong iron chelator that depletes iron from the culture medium. This study finds that adding FeSO4 to a medium that contains PGG restores the biofilm formation and the production of PIA by S. aureus SA113. The requirement of iron for biofilm formation by S. aureus SA113 can also be verified using a semi-defined medium, BM, that contains an iron chelating agent, 2, 2′-dipyridyl (2-DP). Similar to the effect of PGG, the addition of 2-DP to BM medium inhibits biofilm formation and adding FeSO4 to BM medium that contains 2-DP restores biofilm formation. This study reveals an important mechanism of biofilm formation by S. aureus SA113

Tubulin Tyrosination Is Required for the Proper Organization and Pathfinding of the Growth Cone

International audienceBACKGROUND: During development, neuronal growth cones integrate diffusible and contact guidance cues that are conveyed to both actin and microtubule (MT) cytoskeletons and ensure axon outgrowth and pathfinding. Although several post-translational modifications of tubulin have been identified and despite their strong conservation among species, their physiological roles during development, especially in the nervous sytem, are still poorly understood. METHODOLOGY/FINDINGS: Here, we have dissected the role of a post-translational modification of the last amino acid of the alpha-tubulin on axonal growth by analyzing the phenotype of precerebellar neurons in Tubulin tyrosin ligase knock-out mice (TTL(-/-)) through in vivo, ex vivo and in vitro analyses. TTL(-/-) neurons are devoid of tyrosinated tubulin. Their pathway shows defects in vivo, ex vivo, in hindbrains open-book preparations or in vitro, in a collagen matrix. Their axons still orient toward tropic cues, but they emit supernumerary branches and their growth cones are enlarged and exhibit an emission of mis-oriented filopodia. Further analysis of the TTL(-/-) growth cone intracellular organization also reveals that the respective localization of actin and MT filaments is disturbed, with a decrease in the distal accumulation of Myosin IIB, as well as a concomitant Rac1 over-activation in the hindbrain. Pharmacological inhibition of Rac1 over-activation in TTL(-/-) neurons can rescue Myosin IIB localization. CONCLUSIONS/SIGNIFICANCE: In the growth cone, we propose that tubulin tyrosination takes part in the relative arrangement of actin and MT cytoskeletons, in the regulation of small GTPases activity, and consequently, in the proper morphogenesis, organization and pathfinding of the growth cone during development

Autocrine Activation of the MET Receptor Tyrosine Kinase in Acute Myeloid Leukemia

Although the treatment of acute myeloid leukemia (AML) has improved significantly, more than half of all patients develop disease that is refractory to intensive chemotherapy. Functional genomics approaches offer a means to discover specific molecules mediating aberrant growth and survival of cancer cells. Thus, using a loss-of-function RNA interference genomic screen, we identified aberrant expression of the hepatocyte growth factor (HGF) as a critical factor in AML pathogenesis. We found HGF expression leading to autocrine activation of its receptor tyrosine kinase, MET, in nearly half of the AML cell lines and clinical samples studied. Genetic depletion of HGF or MET potently inhibited the growth and survival of HGF-expressing AML cells. However, leukemic cells treated with the specific MET kinase inhibitor crizotinib developed resistance due to compensatory upregulation of HGF expression, leading to restoration of MET signaling. In cases of AML where MET is coactivated with other tyrosine kinases, such as fibroblast growth factor receptor 1 (FGFR1), concomitant inhibition of FGFR1 and MET blocked compensatory HGF upregulation, resulting in sustained logarithmic cell kill both in vitro and in xenograft models in vivo. Our results demonstrate widespread dependence of AML cells on autocrine activation of MET, as well as the importance of compensatory upregulation of HGF expression in maintaining leukemogenic signaling by this receptor. We anticipate that these findings will lead to the design of additional strategies to block adaptive cellular responses that drive compensatory ligand expression as an essential component of the targeted inhibition of oncogenic receptors in human cancers

Metagenomes of the Picoalga Bathycoccus from the Chile Coastal Upwelling

Among small photosynthetic eukaryotes that play a key role in oceanic food webs, picoplanktonic Mamiellophyceae such as Bathycoccus, Micromonas, and Ostreococcus are particularly important in coastal regions. By using a combination of cell sorting by flow cytometry, whole genome amplification (WGA), and 454 pyrosequencing, we obtained metagenomic data for two natural picophytoplankton populations from the coastal upwelling waters off central Chile. About 60% of the reads of each sample could be mapped to the genome of Bathycoccus strain from the Mediterranean Sea (RCC1105), representing a total of 9 Mbp (sample T142) and 13 Mbp (sample T149) of non-redundant Bathycoccus genome sequences. WGA did not amplify all regions uniformly, resulting in unequal coverage along a given chromosome and between chromosomes. The identity at the DNA level between the metagenomes and the cultured genome was very high (96.3% identical bases for the three larger chromosomes over a 360 kbp alignment). At least two to three different genotypes seemed to be present in each natural sample based on read mapping to Bathycoccus RCC1105 genome