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In recent years, porous or sponge based elastomers have been considered to be very useful for wearable applications; however, the one step, economical and rapid fabrication of large area porous elastomers is still a daunting task. In this pursuit, we report a leavening agent mediated fabrication of large area, 3D porous polydimethylsiloxane (PDMS) via a rapid (20 min), cost effective and single step method. The leavening agent used decomposes itself during the leavening process by its inherent autolysis property providing a single step method. A unique feature of this method producing highly uniform and robust 3D porous features is that it can facilitate tuning of sensitivity along with obtaining sponges of desired size and shape. The piezocapacitive devices fabricated using these 3D porous PDMS sponges exhibited exceptional sensitivities of 0.756 kPa(-1) (in the linear region) and 0.0117 kPa(-1) (in the saturation region). These sensors demonstrated an excellent response in small scale as well as large scale motion sensing applications such as pulse pressure sensing, tactile sensing, touch pads, and wrist and palm movements.

Despite the wide application of quantum dots (QDs) in electronics, pharmacy and nanomedicine, limited data is available on their environmental health risk. To advance our current understanding of the environmental impact of these engineered nanomaterials, the aimof this review is to give a detailed insight on the existing information concerning the behaviour, transformation and fate of QDs in the aquatic environment, as well as on its mode of action (MoA), ecotoxicity, trophic transfer and biomagnification at various trophic levels (micro-organisms, aquatic invertebrates and vertebrates). Data show that several types of Cd-based QDs, even at low concentrations (bmg Cd L-1), induce different toxic effects compared to their dissolved counterpart, indicating nano-specific ecotoxicity. QD ecotoxicity at different trophic levels is highly dependent on its physico-chemical properties, environmental conditions, concentration and exposure time, as well as, species, while UV irradiation increases its toxicity. The state of the art regarding the MoA of QDs according to taxonomic groups is summarised and illustrated. Accumulation and trophic transfer of QDs was observed in freshwater and seawater species, while limited biomagnification and detoxification processes were detected. Finally, current knowledge gaps are discussed and recommendations for future research identified. Overall, the knowledge available indicates that in order to develop sustainable nanotechnologies there is an urgent need to develop Cd-free QDs and new "core-shell-conjugate" QD structures. (C) 2016 Elsevier Ltd. All rights reserved. Science Without Borders Program from the Brazilian National Council for Scientific and Technological Development (CNPq) [239524/2012-8] Postdoctoral National Program (PNPD) from the Coordination for the Improvement of Higher Education Personnel (CAPES) Portuguese Foundation for Science and Technology (FCT) project (NANOECOTOX) [PTDC/AAC-AMB/121650/2010] info:eu-repo/semantics/publishedVersion

Cells constantly adapt to environmental fluctuations. These physiological changes require time and therefore cause a lag phase during which the cells do not function optimally. Interestingly, past exposure to an environmental condition can shorten the time needed to adapt when the condition re-occurs, even in daughter cells that never directly encountered the initial condition. Here, we use the molecular toolbox of Saccharomyces cerevisiae to systematically unravel the molecular mechanism underlying such history-dependent behavior in transitions between glucose and maltose. In contrast to previous hypotheses, the behavior does not depend on persistence of proteins involved in metabolism of a specific sugar. Instead, presence of glucose induces a gradual decline in the cells' ability to activate respiration, which is needed to metabolize alternative carbon sources. These results reveal how trans-generational transitions in central carbon metabolism generate history-dependent behavior in yeast, and provide a mechanistic framework for similar phenomena in other cell types. ispartof: ELIFE vol:7 ispartof: location:England status: published

SummaryArchaeal membrane lipid composition is distinct from Bacteria and Eukarya, consisting of isoprenoid chains etherified to the glycerol carbons. Biosynthesis of these lipids is poorly understood. Here we identify and characterize the archaeal membrane protein CDP-archaeol synthase (CarS) that catalyzes the transfer of the nucleotide to its specific archaeal lipid substrate, leading to the formation of a CDP-activated precursor (CDP-archaeol) to which polar head groups are attached. The discovery of CarS enabled reconstitution of the entire archaeal lipid biosynthesis pathway in vitro, starting from simple isoprenoid building blocks and using a set of five purified enzymes. The cell free synthetic strategy for archaeal lipids we describe opens opportunity for studies of archaeal lipid biochemistry. Additionally, insights into archaeal lipid biosynthesis reported here allow addressing the evolutionary hypothesis of the lipid divide between Archaea and Bacteria.

Deconjugation of the Atg8/LC3 protein family members from phosphatidylethanolamine (PE) by Atg4 proteases is essential for autophagy progression, but how this event is regulated remains to be understood. Here, we show that yeast Atg4 is recruited onto autophagosomal membranes by direct binding to Atg8 via two evolutionarily conserved Atg8 recognition sites, a classical LC3-interacting region (LIR) at the C-terminus of the protein and a novel motif at the N-terminus. Although both sites are important for Atg4-Atg8 interaction in vivo, only the new N-terminal motif, close to the catalytic center, plays a key role in Atg4 recruitment to autophagosomal membranes and specific Atg8 deconjugation. We thus propose a model where Atg4 activity on autophagosomal membranes depends on the cooperative action of at least two sites within Atg4, in which one functions as a constitutive Atg8 binding module, while the other has a preference toward PE-bound Atg8.

26 p.-1 fig.-1 tab. Galluzzi, Lorenzo et al., Over the past two decades, the molecular machinery that underlies autophagic responses has been characterized with ever increasing precision in multiple model organisms. Moreover, it has become clear that autophagy and autophagy-related processes have profound implications for human pathophysiology. However, considerable confusion persists about the use of appropriate terms to indicate specific types of autophagy and some components of the autophagy machinery, which may have detrimental effects on the expansion of the field. Driven by the overt recognition of such a potential obstacle, a panel of leading experts in the field attempts here to define several autophagy-related terms based on specific biochemical features. The ultimate objective of this collaborative exchange is to formulate recommendations that facilitate the dissemination of knowledge within and outside the field of autophagy research.

A random library of Escherichia coli MG1655 genomic fragments fused to a promoterless green fluorescent protein (GFP) gene was constructed and screened by differential fluorescence induction for promoters that are induced after exposure to a sublethal high hydrostatic pressure stress. This screening yielded three promoters of genes belonging to the heat shock regulon ( dnaK , lon , clpPX ), suggesting a role for heat shock proteins in protection against, and/or repair of, damage caused by high pressure. Several further observations provide additional support for this hypothesis: (i) the expression of rpoH , encoding the heat shock-specific sigma factor σ 32 , was also induced by high pressure; (ii) heat shock rendered E. coli significantly more resistant to subsequent high-pressure inactivation, and this heat shock-induced pressure resistance followed the same time course as the induction of heat shock genes; (iii) basal expression levels of GFP from heat shock promoters, and expression of several heat shock proteins as determined by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins extracted from pulse-labeled cells, was increased in three previously isolated pressure-resistant mutants of E. coli compared to wild-type levels.

Genetic transformation is a natural process during which foreign DNA enters a cell and integrates into the genome. Apart from its relevance for horizontal gene transfer in nature, transformation is also the cornerstone of today's recombinant gene technology. Despite its importance, relatively little is known about the factors that determine transformation efficiency. We hypothesize that differences in DNA accessibility associated with nucleosome positioning may affect local transformation efficiency. We investigated the landscape of transformation efficiency at various positions in the Saccharomyces cerevisiae genome and correlated these measurements with nucleosome positioning. We find that transformation efficiency shows a highly significant inverse correlation with relative nucleosome density. This correlation was lost when the nucleosome pattern, but not the underlying sequence was changed. Together, our results demonstrate a novel role for nucleosomes and also allow researchers to predict transformation efficiency of a target region and select spots in the genome that are likely to yield higher transformation efficiency. ispartof: Nucleic Acids Research vol:40 issue:19 pages:9506-9512 ispartof: location:England status: published

A typical antioxidant, N-acetyl-L-cysteine (NAC) generally protects cells from oxidative damage induced by reactive oxygen species (ROS). 9,10-Phenanthrenequinone (9,10-PQ), a major quinone in diesel exhaust particles, produces ROS in redox cycling following two-electron reduction by NAD(P)H:quinone oxidoreductase 1 (NQO1), which has been considered as a cause of its cyto- and genotoxicity. In this study, we show that NAC unexpectedly augments the toxicity of 9,10-PQ in cells with low NQO1 activity. In four human skin cell lines, the expression and the activity of NQO1 were lower than in human adenocarcinoma cell lines, A549 and MCF7. In the skin cells, the cytotoxicity of 9,10-PQ was significantly enhanced by addition of NAC. The formation of DNA double strand breaks accompanying phosphorylation of histone H2AX, was also remarkably augmented. On the other hand, the cyto- and genotoxicity were suppressed by addition of NAC in the adenocarcinoma cells. Two contrasting experiments: overexpression of NQO1 in CHO-K1 cells which originally expressed low NQO1 levels, and knock-down of NQO1 in the adenocarcinoma cell line A549 by transfection of RNAi, also showed that NAC suppressed 9,10-PQ-induced toxicity in cell lines expressing high NQO1 activity and enhanced it in cell lines with low NQO1 activity. The results suggested that dual effects of NAC on the cyto- and genotoxicity of 9,10-PQ were dependent on tissue-specific NQO1 activity.

Tolerance to high levels of ethanol is an ecologically and industrially relevant phenotype of microbes, but the molecular mechanisms underlying this complex trait remain largely unknown. Here, we use long-term experimental evolution of isogenic yeast populations of different initial ploidy to study adaptation to increasing levels of ethanol. Whole-genome sequencing of more than 30 evolved populations and over 100 adapted clones isolated throughout this two-year evolution experiment revealed how a complex interplay of de novo single nucleotide mutations, copy number variation, ploidy changes, mutator phenotypes, and clonal interference led to a significant increase in ethanol tolerance. Although the specific mutations differ between different evolved lineages, application of a novel computational pipeline, PheNetic, revealed that many mutations target functional modules involved in stress response, cell cycle regulation, DNA repair and respiration. Measuring the fitness effects of selected mutations introduced in non-evolved ethanol-sensitive cells revealed several adaptive mutations that had previously not been implicated in ethanol tolerance, including mutations in PRT1, VPS70 and MEX67. Interestingly, variation in VPS70 was recently identified as a QTL for ethanol tolerance in an industrial bio-ethanol strain. Taken together, our results show how, in contrast to adaptation to some other stresses, adaptation to a continuous complex and severe stress involves interplay of different evolutionary mechanisms. In addition, our study reveals functional modules involved in ethanol resistance and identifies several mutations that could help to improve the ethanol tolerance of industrial yeasts., Author Summary Organisms can evolve resistance to specific stress factors, which allows them to thrive in environments where non-adapted organisms fail to grow. However, the molecular mechanisms that underlie adaptation to complex stress factors that interfere with basic cellular processes are poorly understood. In this study, we reveal how yeast populations adapt to high ethanol concentrations, an ecologically and industrially relevant stress that is still poorly understood. We exposed six independent populations of genetically identical yeast cells to gradually increasing ethanol levels, and we monitored the changes in their DNA sequence over a two-year period. Together with novel computational analyses, we could identify the mutational dynamics and molecular mechanisms underlying increased ethanol resistance. Our results show how adaptation to high ethanol is complex and can be reached through different mutational pathways. Together, our study offers a detailed picture of how populations adapt to a complex continuous stress and identifies several mutations that increase ethanol resistance, which opens new routes to obtain superior biofuel yeast strains.

Rice blast fungus ( Magnaporthe grisea) forms a highly specialized infection structure for plant penetration, the appressorium, the formation and growth of which are regulated by the Mst11- Mst7- Pmk1 mitogen- activated protein kinase cascade. We characterized the MST50 gene that directly interacts with both MST11 and MST7. Similar to the mst11 mutant, the mst50 mutant was defective in appressorium formation, sensitive to osmotic stresses, and nonpathogenic. Expressing a dominant active MST7 allele in mst50 complemented its defects in appressorium but not lesion formation. The sterile alpha-motif ( SAM) domain of Mst50 was essential for its interaction with Mst11 and for appressorium formation. Although the SAM and Ras-association domain ( RAD) of Mst50 were dispensable for its interaction with Mst7, deletion of RAD reduced appressorium formation and virulence on rice ( Oryza sativa) seedlings. The interaction between Mst50 and Mst7 or Mst11 was detected by coimmunoprecipitation assays in developing appressoria. Mst50 also interacts with Ras1, Ras2, Cdc42, and Mgb1 in yeast two- hybrid assays. Expressing a dominant active RAS2 allele in the wild- type strain but not in mst50 stimulated abnormal appressorium formation. These results indicate that MST50 functions as an adaptor protein interacting with multiple upstream components and plays critical roles in activating the Pmk1 cascade for appressorium formation and plant infection in M. grisea.

The organization of multicellular organisms depends on cell-cell communication. The signal molecules are often soluble components in the extracellular fluid, but also include odors and light. A large array of surface receptors is involved in the detection of these signals. Signals are then transduced across the plasma membrane so that enzymes at the inner face of the membrane are activated, producing second messengers, which by a complex network of interactions activate target proteins or genes. Vertebrate cells have been used to study hormone and neurotransmitter action, vision, the regulation of cell growth and differentiation. Sensory transduction in lower eukaryotes is predominantly used for other functions, notably cell attraction for mating and food seeking. By comparing sensory transduction in lower and higher eukaryotes general principles may be recognized that are found in all organisms and deviations that are present in specialised systems. This may also help to understand the differences between cell types within one organism and the importance of a particular pathway that may or may not be general. In a practical sense, microorganisms have the advantage of their easy genetic manipulation, which is especially advantageous for the identification of the function of large families of signal transducing components.