A gene expression study on strains of Nostoc (Cyanobacteria) revealing antimicrobial activity under mixotrophic conditions

Cyanobacteria are well known for their production of a multitude of highly allelopathic compounds. These products have features such as incorporation of non-proteinogenic amino acids which are characteristics of peptides biosynthesized by non-ribosomal peptide synthetases (NRPSs). Some of these peptides have acetate-derived moieties, suggesting that their biosynthesis also involves polyketide synthases (PKSs). Among the photosynthetic microorganisms, cyanobacteria belonging to the genus Nostoc are regarded as good candidates for producing biologically active secondary metabolites. Aiming at the maximization in the production of natural product, we compared autotrophic, and mixotrophic growth at high light intensity of two Nostoc species in relation to the production of bioactive compounds with the antimicrobial activity at different source of sugar. Glucose was shown to be the best substrate for the production of high natural product when compared with sucrose. Also, the rate of biomass production and antimicrobial activity was reaching ~2.0 to 2.5 and ~1.5 times greater than that of the autotrophic and sucrose grown cultures, respectively. Also, we conduct a combined NRPSs and PKSs polymerase chain reaction (PCR). The sequences presented in this study was deposited in GenBank and had accession numbers JF795278 and JF795279 (NRPS A domains) and JF795280 and JF795281 (PKS KS domains). Computer modeling and phylogenetic analysis was conducted to predict the putative amino acid recognized by the unknown adenylation domain in the NRPS sequences. This study highlights the importance of environmental and nutrimental factors in maximization of antibiotic production of two Nostoc species.Keywords: Peptide synthetase gene, polyketide synthase gene, Nostoc, secondary metabolites, mixotrophic condition

The Effects of Excess Copper on Antioxidative Enzymes, Lipid Peroxidation, Proline, Chlorophyll, and Concentration of Mn, Fe, and Cu in Astragalus neo-mobayenii

To probe the physiological and biochemical tolerance mechanisms in Astragalus neo-mobayenii Maassoumi, an endemic plant around the Cu-rich areas from the North West of Iran, the effect of different copper concentrations at toxic levels on this plant was investigated. Copper was applied in the form of copper sulfate (CuSO4·5H2O) in four levels (0, 50, 100, and 150 μM). We observed no visible symptoms of Cu toxicity in this plant species. During the exposure of plants to excess copper, the antioxidant defense system helped the plant to protect itself from the damage. With increasing copper concentration, superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities increased in leaves and roots () compared with that of the control group. The chlorophyll amount gradually declined with increasing Cu concentrations. However, reduction in the 50 μM level showed insignificant changes. Enhanced accumulation of proline content in the leaves was determined, as well as an increase of MDA content (oxidative damage biomarker) (). The results indicated that Cu contents in leaves and roots enhanced with increasing levels of Cu application. The Fe and Mn contents in both shoots and roots significantly decreased with increasing Cu concentration. Finally, the mechanisms of copper toxicity and copper tolerance in this plant were briefly discussed

Deficit of wide binaries in the eta Chamaeleontis young cluster

We have carried out a sensitive high-resolution imaging survey of stars in the young (6-8 Myr), nearby (97 pc) compact cluster around eta Chamaeleontis to search for stellar and sub-stellar companions. Given its youth and proximity, any sub-stellar companions are expected to be luminous, especially in the near infrared, and thus easier to detect next to their parent stars. Here, we present VLT/NACO adaptive optics imaging with companion detection limits for 17 eta Cha cluster members, and follow-up VLT/ISAAC near-infrared spectroscopy for companion candidates. The widest binary detected is ~0.2", corresponding to the projected separation 20 AU, despite our survey being sensitive down to sub-stellar companions outside 0.3", and planetary mass objects outside 0.5". This implies that the stellar companion probability outside 0.3" and the brown dwarf companion probability outside 0.5" are less than 0.16 with 95% confidence. We compare the wide binary frequency of eta Cha to that of the similarly aged TW Hydrae association, and estimate the statistical likelihood that the wide binary probability is equal in both groups to be < 2e-4. Even though the eta Cha cluster is relatively dense, stellar encounters in its present configuration cannot account for the relative deficit of wide binaries. We thus conclude that the difference in wide binary probability in these two groups provides strong evidence for multiplicity properties being dependent on environment. In two appendices we derive the projected separation probability distribution for binaries, used to constrain physical separations from observed projected separations, and summarize statistical tools useful for multiplicity studies.Comment: Accepted by ApJ. 13 pages, 10 figure

Topical Gene Electrotransfer to the Epidermis of Hairless Guinea Pig by Non-invasive Multielectrode Array

Topical gene delivery to the epidermis has the potential to be an effective therapy for skin disorders, cutaneous cancers, vaccinations and systemic metabolic diseases. Previously, we reported on a non-invasive multielectrode array (MEA) that efficiently delivered plasmid DNA and enhanced expression to the skin of several animal models by in vivo gene electrotransfer. Here, we characterized plasmid DNA delivery with the MEA in a hairless guinea pig model, which has a similar histology and structure to human skin. Significant elevation of gene expression up to 4 logs was achieved with intradermal DNA administration followed by topical non-invasive skin gene electrotransfer. This delivery produced gene expression in the skin of hairless guinea pig up to 12 to 15 days. Gene expression was observed exclusively in the epidermis. Skin gene electrotransfer with the MEA resulted in only minimal and mild skin changes. A low level of human Factor IX was detected in the plasma of hairless guinea pig after geneelectrotransfer with the MEA, although a significant increase of Factor IX was obtained in the skin of animals. These results suggest geneelectrotransfer with the MEA can be a safe, efficient, non-invasive skin delivery method for skin disorders, vaccinations and potential systemic diseases where low levels of gene products are sufficient

Determinants of a transcriptionally competent environment at the GM-CSF promoter

Granulocyte macrophage-colony stimulating factor (GM-CSF) is produced by T cells, but not B cells, in response to immune signals. GM-CSF gene activation in response to T-cell stimulation requires remodelling of chromatin associated with the gene promoter, and these changes do not occur in B cells. While the CpG methylation status of the murine GM-CSF promoter shows no correlation with the ability of the gene to respond to activation, we find that the basal chromatin environment of the gene promoter influences its ability to respond to immune signals. In unstimulated T cells but not B cells, the GM-CSF promoter is selectively marked by enrichment of histone acetylation, and association of the chromatin-remodelling protein BRG1. BRG1 is removed from the promoter upon activation concomitant with histone depletion and BRG1 is required for efficient chromatin remodelling and transcription. Increasing histone acetylation at the promoter in T cells is paralleled by increased BRG1 recruitment, resulting in more rapid chromatin remodelling, and an associated increase in GM-CSF mRNA levels. Furthermore, increasing histone acetylation in B cells removes the block in chromatin remodelling and transcriptional activation of the GM-CSF gene. These data are consistent with a model in which histone hyperacetylation and BRG1 enrichment at the GM-CSF promoter, generate a chromatin environment competent to respond to immune signals resulting in gene activation

An integrated pan-European research infrastructure for validating smart grid systems

A driving force for the realization of a sustainable energy supply in Europe is the integration of distributed, renewable energy resources. Due to their dynamic and stochastic generation behaviour, utilities and network operators are confronted with a more complex operation of the underlying distribution grids. Additionally, due to the higher flexibility on the consumer side through partly controllable loads, ongoing changes of regulatory rules, technology developments, and the liberalization of energy markets, the system’s operation needs adaptation. Sophisticated design approaches together with proper operational concepts and intelligent automation provide the basis to turn the existing power system into an intelligent entity, a so-called smart grid. While reaping the benefits that come along with those intelligent behaviours, it is expected that the system-level testing will play a significantly larger role in the development of future solutions and technologies. Proper validation approaches, concepts, and corresponding tools are partly missing until now. This paper addresses these issues by discussing the progress in the integrated Pan-European research infrastructure project ERIGrid where proper validation methods and tools are currently being developed for validating smart grid systems and solutions.This work is supported by the European Community’s Horizon 2020 Program (H2020/2014-2020) under project “ERIGrid” (Grant Agreement No. 654113). Further information is available at the corresponding website www.erigrid.eu

A novel mechanism for target gene-specific SWI/SNF recruitment via the Snf2p N-terminus

Chromatin-remodeling complexes regulate the expression of genes in all eukaryotic genomes. The SWI/SNF complex of Saccharomyces cerevisiae is recruited to its target promoters via interactions with selected transcription factors. Here, we show that the N-terminus of Snf2p, the chromatin remodeling core unit of the SWI/SNF complex, is essential for the expression of VHT1, the gene of the plasma membrane H+/biotin symporter, and of BIO5, the gene of a 7-keto-8-aminopelargonic acid transporter, biotin biosynthetic precursor. chromatin immunoprecipitation (ChIP) analyses demonstrate that Vhr1p, the transcriptional regulator of VHT1 and BIO5 expression, is responsible for the targeting of Snf2p to the VHT1 promoter at low biotin. We identified an Snf2p mutant, Snf2p-R15C, that specifically abolishes the induction of VHT1 and BIO5 but not of other Snf2p-regulated genes, such as GAL1, SUC2 or INO1. We present a novel mechanism of target gene-specific SWI/SNF recruitment via Vhr1p and a conserved N-terminal Snf2p domain

Lineage-specific dynamic and pre-established enhancer–promoter contacts cooperate in terminal differentiation

Chromosome conformation is an important feature of metazoan gene regulation; however, enhancer–promoter contact remodeling during cellular differentiation remains poorly understood. To address this, genome-wide promoter capture Hi-C (CHi-C) was performed during epidermal differentiation. Two classes of enhancer–promoter contacts associated with differentiation-induced genes were identified. The first class ('gained') increased in contact strength during differentiation in concert with enhancer acquisition of the H3K27ac activation mark. The second class ('stable') were pre-established in undifferentiated cells, with enhancers constitutively marked by H3K27ac. The stable class was associated with the canonical conformation regulator cohesin, whereas the gained class was not, implying distinct mechanisms of contact formation and regulation. Analysis of stable enhancers identified a new, essential role for a constitutively expressed, lineage-restricted ETS-family transcription factor, EHF, in epidermal differentiation. Furthermore, neither class of contacts was observed in pluripotent cells, suggesting that lineage-specific chromatin structure is established in tissue progenitor cells and is further remodeled in terminal differentiation

MicroRNA-21 targets tumor suppressor genes ANP32A and SMARCA4

MicroRNA-21 (miR-21) is a key regulator of oncogenic processes. It is significantly elevated in the majority of human tumors and functionally linked to cellular proliferation, survival and migration. In this study, we used two experimental-based strategies to search for novel miR-21 targets. On the one hand, we performed a proteomic approach using two-dimensional differential gel electrophoresis (2D-DIGE) to identify proteins suppressed upon enhanced miR-21 expression in LNCaP human prostate carcinoma cells. The tumor suppressor acidic nuclear phosphoprotein 32 family, member A (ANP32A) (alias pp32 or LANP) emerged as the most strongly downregulated protein. On the other hand, we applied a mathematical approach to select correlated gene sets that are negatively correlated with primary-miR-21 (pri-miR-21) expression in published transcriptome data from 114 B-cell lymphoma cases. Among these candidates, we found tumor suppressor SMARCA4 (alias BRG1) together with the already validated miR-21 target, PDCD4. ANP32A and SMARCA4, which are both involved in chromatin remodeling processes, were confirmed as direct miR-21 targets by immunoblot analysis and reporter gene assays. Furthermore, knock down of ANP32A mimicked the effect of enforced miR-21 expression by enhancing LNCaP cell viability, whereas overexpression of ANP32A in the presence of high miR-21 levels abrogated the miR-21-mediated effect. In A172 glioblastoma cells, enhanced ANP32A expression compensated for the effects of anti-miR-21 treatment on cell viability and apoptosis. In addition, miR-21 expression clearly increased the invasiveness of LNCaP cells, an effect also seen in part upon downregulation of ANP32A. In conclusion, these results suggest that downregulation of ANP32A contributes to the oncogenic function of miR-21