Self-Assembling Systems Based on Pillar[5]arenes and Surfactants for Encapsulation of Diagnostic Dye DAPI

In this paper, we report the development of the novel self-assembling systems based on oppositely charged Pillar[5]arenes and surfactants for encapsulation of diagnostic dye DAPI. For this purpose, the aggregation behavior of synthesized macrocycles and surfactants in the presence of Pillar[5]arenes functionalized by carboxy and ammonium terminal groups was studied. It has been demonstrated that by varying the molar ratio in Pillar[5]arene-surfactant systems, it is possible to obtain various types of supramolecular systems: host–guest complexes at equimolar ratio of Pillar[5]arene-surfactant and interpolyelectrolyte complexes (IPECs) are self-assembled materials formed in aqueous medium by two oppositely charged polyelectrolytes (macrocycle and surfactant micelles). It has been suggested that interaction of Pillar[5]arenes with surfactants is predominantly driven by cooperative electrostatic interactions. Synthesized stoichiometric and non-stoichiometric IPECs specifically interact with DAPI. UV-vis, luminescent spectroscopy and molecular docking data show the structural feature of dye-loaded IPEC and key role of the electrostatic, π–π-stacking, cation–π interactions in their formation. Such a strategy for the design of supramolecular Pillar[5]arene-surfactant systems will lead to a synergistic interaction of the two components and will allow specific interaction with the third component (drug or fluorescent tag), which will certainly be in demand in pharmaceuticals and biomedical diagnostics.     

Sustainable Hydrogen Photoproduction by Phosphorus-Deprived Marine Green Microalgae Chlorella sp.

Previously it has been shown that green microalga Chlamydomonas reinhardtii is capable of prolonged H₂ photoproduction when deprived of sulfur. In addition to sulfur deprivation (-S), sustained H₂ photoproduction in C. reinhardtii cultures can be achieved under phosphorus-deprived (-P) conditions. Similar to sulfur deprivation, phosphorus deprivation limits O₂ evolving activity in algal cells and causes other metabolic changes that are favorable for H₂ photoproduction. Although significant advances in H₂ photoproduction have recently been realized in fresh water microalgae, relatively few studies have focused on H₂ production in marine green microalgae. In the present study phosphorus deprivation was applied for hydrogen production in marine green microalgae Chlorella sp., where sulfur deprivation is impossible due to a high concentration of sulfates in the sea water. Since resources of fresh water on earth are limited, the possibility of hydrogen production in seawater is more attractive. In order to achieve H₂ photoproduction in P-deprived marine green microalgae Chlorella sp., the dilution approach was applied. Cultures diluted to about 0.5–1.8 mg Chl·L⁻¹ in the beginning of P-deprivation were able to establish anaerobiosis, after the initial growth period, where cells utilize intracellular phosphorus, with subsequent transition to H₂ photoproduction stage. It appears that marine microalgae during P-deprivation passed the same stages of adaptation as fresh water microalgae. The presence of inorganic carbon was essential for starch accumulation and subsequent hydrogen production by microalgae. The H₂ accumulation was up to 40 mL H₂ gas per 1iter of the culture, which is comparable to that obtained in P-deprived C. reinhardtii culture.     

A model for European Biogenic Volatile Organic Compound emissions: Software development and first validation

A grid-oriented Biogenic Emission Model (BEM) has been developed to calculate Non-Methane Volatile Organic Compound (NMVOC) emissions from vegetation in high spatial and temporal resolutions. The model allows the emissions calculation for any modeling domain covering Europe on the basis of: 1) the U.S. Geological Survey 1-km resolution land-use database, 2) a land-use specific, monthly isoprene, monoterpene and Other Volatile Organic Compound (OVOC) emission potentials and foliar biomass densities database, 3) temperature and solar radiation data provided by the mesoscale meteorological model MM5. The model was applied for Europe in 30-km spatial resolution for the year 2003. The European total emissions for 2003 consist of 33.0% isoprene, 25.5% monoterpenes and 41.5% OVOC. BEM results are compared with those from the well-documented global Model of Emissions of Gases and Aerosols from Nature (MEGAN). The BEM total emissions compare well with the MEGAN ones. In July 2003, the results of both models agree within a factor of 1.2 for total isoprene emissions and within a factor of 2 for total monoterpene emissions. The comparison of the spatial distributions of the July 2003 isoprene and monoterpene emissions calculated with BEM and MEGAN shows that, in the greater part of the study area, the differences are below the current uncertainty limit for the estimation of spatially-resolved biogenic VOC emissions in Europe being equal to about ±600 kg km^?2 month^?1. Differences that are above this limit are found mainly in the eastern European countries for isoprene and in the Mediterranean countries for monoterpenes.     

Subcellular distribution and function of Rab3A, B, C, and D isoforms in insulin-secreting cells

Insulin-secreting cells express four GTPases of the Rab3 family. After separation of extracts of INS-1 cells on a sucrose density gradient, the bulk of the A, B, and C isoforms was recovered in the fractions enriched in insulin-containing secretory granules. Rab3D was also mainly associated with secretory granules, but a fraction of this isoform was localized on lighter organelles. Analyses by confocal microscopy of immunostained HIT-T15 cells transfected with epitope-tagged constructs confirmed the distribution of the Rab3 isoforms. Transfection of HIT-T15 cells with GTPase-deficient mutants of the Rab3 isoforms decreased nutrient-induced insulin release to different degrees (D>B>A>C), while overexpression of Rab3 wild types had minor or no effects. Expression of the same Rab3 mutants in PC12 cells provoked an inhibition of K+-stimulated secretion of dense core vesicles, indicating that, in beta-cells and neuroendocrine cells, the four Rab3 isoforms play a similar role in exocytosis. A Rab3A/C chimera in which the carboxyterminal domain of A was replaced with the corresponding region of C inhibited insulin secretion as Rab3A. In contrast, a Rab3C/A chimera containing the amino-terminal domain of C was less potent and reduced exocytosis as Rab3C. This suggests that the degree of inhibition obtained after transfection of the Rab3 isoforms is determined by differences in the variable amino-terminal region.     

Flux balance analysis of photoheterotrophic growth of purple nonsulfur bacteria relevant to biohydrogen production

Purple nonsulfur bacteria can produce hydrogen during photoheterotrophic growth on a variety of organic substrates. To facilitate metabolic engineering of this group of bacteria with the purpose of maximizing hydrogen production, we have developed the first comprehensive mathematical model of anaerobic photoheterotrophic metabolism. This model is a stoichiometric model to which Flux Balance Analysis also known as constraint-based analysis has been applied. Because Rhodobacter sphaeroides emerged as one of the best understood purple nonsulfur bacteria, we based our modeling on this species. However, the approach is readily applicable to other purple nonsulfur bacteria. The metabolic space used in model construction contains 314 reconstructed biochemical reactions and 287 compounds. The list of simulated metabolic pathways involves major carbon and nitrogen metabolic pathways. A distinctive feature of our model is that it considers maximization of two different cell objective functions, namely, biomass production rate, which is commonly used in flux balance models, and reductant removal rate, which is a unique feature of this model reflecting peculiarity of photoheterotrophic metabolism. The performance of this model has been assessed by comparing computed metabolic fluxes with experimental values obtained by several research groups who worked on various strains of purple nonsulfur bacteria and utilized different growth setups. The model testing revealed remarkably good agreement between the predicted and measured metabolic fluxes, which validates the presented model.     

Remodeling of Arterial Tone Regulation in Postnatal Development: Focus on Smooth Muscle Cell Potassium Channels

Maturation of the cardiovascular system is associated with crucial structural and functional remodeling. Thickening of the arterial wall, maturation of the sympathetic innervation, and switching of the mechanisms of arterial contraction from calcium-independent to calcium-dependent occur during postnatal development. All these processes promote an almost doubling of blood pressure from the moment of birth to reaching adulthood. This review focuses on the developmental alterations of potassium channels functioning as key smooth muscle membrane potential determinants and, consequently, vascular tone regulators. We present evidence that the pattern of potassium channel contribution to vascular control changes from K_ir2, K_v1, K_v7 and TASK-1 channels to BK_Ca channels with maturation. The differences in the contribution of potassium channels to vasomotor tone at different stages of postnatal life should be considered in treatment strategies of cardiovascular diseases associated with potassium channel malfunction.     

Deformation in viscoelastic sandwich composites subject to moisture diffusion

This study analyzes the effect of moisture diffusion on the deformation of viscoelastic sandwich composites, which are composed of orthotropic fiber-reinforced laminated skins and viscoelastic polymeric foam core. It is assumed that the elastic and time-dependent (transient) moduli at any particular location in the foam core depend on the moisture concentration at that location. Sequentially coupled analyses of moisture diffusion and deformation are performed to predict overall performance of the studied viscoelastic sandwich systems. Time and moisture dependent constitutive model is used for the polymer foam core, while skins are assumed linear elastic. The overall time-dependent responses of the sandwich composites subject to moisture diffusion are analyzed using finite element (FE) method. Experimental data available in the literature and analytical solutions are used to support convergence studies in the FE analyses. Contributions of moisture dependent elastic and the time-dependent moduli to the overall stress, strain and displacement field are studied. FE analyses of the delamination between skins and core in sandwich composite under combined moisture diffusion and mechanical loading are also performed.     

Carrageenan-based superabsorbent biopolymers mitigate autogenous shrinkage in ordinary portland cement

We report the synthesis and characterization of biobased superabsorbent copolymers from κ-carrageenan and poly(acrylic acid) that mitigate autogenous shrinkage in ordinary portland cement paste. Synthesized via free radical graft polymerization, the biobased superabsorbent polymers (SAPs) were characterized with regard to their thermochemical properties and swelling behavior in both aqueous and ionic solutions. The biobased SAPs were incorporated into cement paste to investigate their ability to mitigate autogenous shrinkage cracking in high-performance concrete. Results demonstrate that the biobased SAPs absorb up to 438 and 94 [g/g] (by mass) in aqueous and ionic solutions, respectively, after 24 h. Furthermore, the biobased SAPs were successful in mitigating shrinkage in low water-to-cement ratio pastes. While the control paste exhibited negative strain and ultimate shrinkage cracking, the samples containing biobased SAP experienced net-positive expansion during cement hydration.     

A study on Dual-Scale data charts

We present the results of a user study that compares different ways of representing Dual-Scale data charts. Dual-Scale charts incorporate two different data resolutions into one chart in order to emphasize data in regions of interest or to enable the comparison of data from distant regions. While some design guidelines exist for these types of charts, there is currently little empirical evidence on which to base their design. We fill this gap by discussing the design space of Dual-Scale cartesian-coordinate charts and by experimentally comparing the performance of different chart types with respect to elementary graphical perception tasks such as comparing lengths and distances. Our study suggests that cut-out charts which include collocated full context and focus are the best alternative, and that superimposed charts in which focus and context overlap on top of each other should be avoided.     

The Functional Role of Loops and Flanking Sequences of G-Quadruplex Aptamer to the Hemagglutinin of Influenza a Virus

Nucleic acid aptamers are generally accepted as promising elements for the specific and high-affinity binding of various biomolecules. It has been shown for a number of aptamers that the complexes with several related proteins may possess a similar affinity. An outstanding example is the G-quadruplex DNA aptamer RHA0385, which binds to the hemagglutinins of various influenza A virus strains. These hemagglutinins have homologous tertiary structures but moderate-to-low amino acid sequence identities. Here, the experiment was inverted, targeting the same protein using a set of related, parallel G-quadruplexes. The 5′- and 3′-flanking sequences of RHA0385 were truncated to yield parallel G-quadruplex with three propeller loops that were 7, 1, and 1 nucleotides in length. Next, a set of minimal, parallel G-quadruplexes with three single-nucleotide loops was tested. These G-quadruplexes were characterized both structurally and functionally. All parallel G-quadruplexes had affinities for both recombinant hemagglutinin and influenza virions. In summary, the parallel G-quadruplex represents a minimal core structure with functional activity that binds influenza A hemagglutinin. The flanking sequences and loops represent additional features that can be used to modulate the affinity. Thus, the RHA0385–hemagglutinin complex serves as an excellent example of the hypothesis of a core structure that is decorated with additional recognizing elements capable of improving the binding properties of the aptamer.