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Ab initio molecular dynamics simulations are reported forwater-embedded TiO2 surfaces to determine the diffusive and reactive behavior at full hydration. A three-domain model is developed for six surfaces [rutile (110), (100), and (001), and anatase (101), (100), and (001)] which describes waters as hard (irreversibly bound to the surface), soft (with reduced mobility but orientation freedom near the surface), or bulk. The model explains previous experimental data and provides a detailed picture of water diffusion near TiO2 surfaces. Water reactivity is analyzed with a graph-theoretic approach that reveals a number of reaction pathways on TiO2 which occur at full hydration, in addition to direct water splitting. Hydronium (H3O+) is identified to be a key intermediate state, which facilitates water dissociation by proton hopping between intact and dissociated waters near the surfaces. These discoveries significantly improve the understanding of nanoscale water dynamics and reactivity at TiO2 interfaces under ambient conditions.
Atomistic simulations are powerful for probing molecules at bioinorganic interfaces and excellent complements to scarcely available experimental techniques. The free energy controls the adsorption behavior of molecules on nanosurfaces, and is therefore a quantity of particular importance. Advanced sampling techniques can efficiently explore the adsorption free energy landscape, but molecular simulations with classical (Newtownian) dynamics fail to capture charge transfer and polarization at the solid-liquid interface. First principle simulations do not suffer from this limitation but come with a heavy computational load. Here, we introduce an efficient protocol to explore the free energy of adsorption in the ab initio framework. This approach accurately models the complex phenomena at bio-inorganic surfaces on the nanoscale and properly samples the relevant thermodynamic properties. We present a case study of adsorption of the Lysine and Aspartate amino acids on the anatase (101) TiO2 surface with the tight binding method. The high values of the calculated adsorption free energies highlight the importance of a proper description of the electronic state for surface binding processes.
Nucleic acids are highly charged biopolymers whose secondary structure is strongly dependent on electrostatic interactions. Solvent molecules and ions are also believed to play an important role in mediating and directing both sequence recognition and interactions with other molecules, such as proteins and a variety of ligands. Therefore, to fully understand the biological functions of DNA, it is necessary to understand the interactions with the surrounding counterions. It is well known that monovalent counterions can bind to the minor groove of DNA with consecutive sequences of four, or more, adenine and thymine (A-tracts) with relatively long residence times. However, much less is known about their binding to the backbone and to the major groove. In this work, we used molecular dynamics simulations to both investigate the interactions between the backbone and major groove of DNA and one of its physiological counterions (Na+) and evaluate the relationship between these interactions and the nucleotide sequence. Three dodecamers, namely CGAAAATTTTCG, CGCTCTAGAGCG, and CGCGAATTCGCG, were simulated using the Toukan-Rahman flexible SPC water model and Smith and Dang parameters for Na+, revealing a significant sequence dependence on the ion binding to both backbone and major groove. In the absence of experimental data on the atomistic details of the studied interactions, the reliability of the results was evaluated performing the simulations with additional sets of potential parameters for ions and solvent, namely the A. qvist or the Joung and Cheatham ion parameters and the TIP3P water model. This allowed us to evaluate the results by verifying which features are preserved independently from the parameters adopted.
Small-sized wet TiO2 nanoparticles have been investigated by ab initio molecular dynamics simulations. Chemical and physical adsorption of water on the TiO2-water interface was studied as a function of water content, ranging from dry nanoparticles to wet nanoparticles with monolayer coverage of water. The surface reactivity was shown to be a concave function of water content and driven by surface defects. The local coordination number at the defect was identified as the key factor to decide whether water adsorption proceeds through dissociation or physisorption on the surface. A consistent picture of TiO2 nanoparticle wetting at the microscopic level emerges, which corroborates existing experimental data and gives further insight into the molecular mechanisms behind nanoparticle wetting. These calculations will facilitate the engineering of metal oxide nanoparticles with a controlled catalytic water activity.
Adsorption profiles and adsorption free energies were determined for the side chain analogues of the 20 naturally occurring amino acids and a titanium binding peptide on the TiO2 (100) surface. Microsecond simulations with umbrella sampling and metadynamics were used to sample the free energy barriers associated with desolvation of strongly bound water molecules at the TiO2 surface. Polar and aromatic side chain analogues that hydrogen bond either to surface waters or directly to the metal oxide surface were found to be the strongest binders. Further, adsorption simulations of a 6 residue titanium binding peptide identified two binding modes on TiO2 (100). The peptide structure with lowest free energy was shown to be stabilized by a salt bridge between the end termini. A comparison between the free energies of the side chain analogues of the peptide sequence and the peptide itself shows that the free energy contributions are not additive. The simulations emphasize that tightly bound surface waters play a key role for peptide and protein structures when bound to inorganic surfaces in biological environments.
Atomistic force field parameters were developed for the TiO2-water interface by systematic optimization with respect to experimentally determined crystal structures (lattice parameters) and surface thermodynamics (water adsorption enthalpy). Optimized force field parameters were determined for the two cases where TiO2 was modeled with or without covalent bonding. The nonbonded TiO2 model can be used to simulate different TiO2 phases, while the bonded TiO2 model is particularly useful for simulations of nanosized TiO2 and biomatter, including protein-surface and nanoparticle-biomembrane simulations. The procedure is easily generalized to parametrize interactions between other inorganic surfaces and biomolecules.
Ionic liquids (ILs) are a class of new green materials that have attracted extensive attention in recent decades. Many novel properties not evident under normal conditions may appear when ionic liquids are confined to a nanometer scale. As was observed in the experiment, an anomalous phase behavior from liquid to high melting point perfect crystal occurred when 1-n-butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF6]) ionic liquid was confined in a carbon nanotube. In this work, we performed molecular dynamics (MD) simulations for [bmim][PF6] ionic liquid and provided direct structural evidence that the ionic crystallizes in a carbon nanotube. The ordered ionic arrangement in both the radial and the axial directions can be observed inside the channels of the CNTs to induce the form of crystallites. The ionic stacking and distributing can be determined by the sizes of the CNTs. Hydrogen bonds remain the dominant interactions between cations and anions when the ionic liquid enters into the CNT from the bulk phase. The free energies as the thermal driven forces were calculated, and it is found that it is very difficult for a single anion to enter into the channel of the CNT spontaneously. A more favorable way is through an ion-pair in which a cation "pulls" an anion to enter into the channel of the CNT together. It is predicted that other ionic liquids that possess similar structures, even including the pyridinium-based ionic liquids, can show higher melting points when confined in CNTs.
To study the effects of water on conformational dynamics of polyalcohols, Molecular Dynamics simulations of glycerol water liquid mixtures have been carried out at different concentrations: 42.9 and 60.0 wt 96 of glycerol, respectively. On the basis of the analysis of backbone conformer distributions, it is found that the surrounding water molecules have a large impact on the populations of the glycerol conformers. While the local structure of water in the liquid mixture is surprisingly close to that in pure liquid water, the behavior of glycerols can be divided into three different categories where roughly 25% of them occur in a structure similar to that in pure liquid of glycerol, ca. 25% of them exist as monomers, solvated by water, and the remaining 50% of glycerols in the mixture form H-bonded strings as. remains of the glycerol H-bond network. The typical glycerol H-bond network still exists even at the lower concentration of 40 wt % of glycerol. The microheterogeneity of water glycerol mixtures is analyzed using time-averaged distributions of the sizes of the water aggregates. At 40 wt % of glycerol, the cluster sizes from 3 to 10 water molecules are observed. The increase of glycerol content causes a depletion of clusters leading to smaller 3-5 molecule clusters domination. Translational diffusion coefficients have been calculated to study the dynamical behavior of both glycerol and water molecules. Rotational-reorientational motion is studied both in overall and in selected substructures on the basis of time correlation functions. Characteristic time scales for different motional modes are deduced on the basis of the calculated correlation times. The general conclusion is that the presence of water increases the overall mobility of glycerol, while glycerol slows the mobility of water.
Cardiolipin is a non-bilayer phospholipid with a unique dimeric structure. It localizes to negative curvature regions in bacteria and is believed to stabilize respiratory chain complexes in the highly curved mitochondrial membrane. Cardiolipin's localization mechanism remains unresolved, because important aspects such as the structural basis and strength for lipid curvature preferences are difficult to determine, partly due to the lack of efficient simulation methods. Here, we report a computational approach to study curvature preferences of cardiolipin by simulated membrane buckling and quantitative modeling. We combine coarse-grained molecular dynamics with simulated buckling to determine the curvature preferences in three-component bilayer membranes with varying concentrations of cardiolipin, and extract curvature-dependent concentrations and lipid acyl chain order parameter profiles. Cardiolipin shows a strong preference for negative curvatures, with a highly asymmetric chain order parameter profile. The concentration profiles are consistent with an elastic model for lipid curvature sensing that relates lipid segregation to local curvature via the material constants of the bilayers. These computations constitute new steps to unravel the molecular mechanism by which cardiolipin senses curvature in lipid membranes, and the method can be generalized to other lipids and membrane components as well.
Membrane curvature sensing, where the binding free energies of membrane-associated molecules depend on the local membrane curvature, is a key factor to modulate and maintain the shape and organization of cell membranes. However, the microscopic mechanisms are not well understood, partly due to absence of efficient simulation methods. Here, we describe a method to compute the curvature dependence of the binding free energy of a membrane associated probe molecule that interacts with a buckled membrane, which has been created by lateral compression of a flat bilayer patch. This buckling approach samples a wide range of curvatures in a single simulation, and anisotropic effects can be extracted from the orientation statistics. We develop an efficient and robust algorithm to extract the motion of the probe along the buckled membrane surface, and evaluate its numerical properties by extensive sampling of three coarse-grained model systems: local lipid density in a curved environment for single-component bilayers, curvature preferences of individual lipids in two-component membranes, and curvature sensing by a homotrimeric transmembrane protein. The method can be used to complement experimental data from curvature partition assays and provides additional insight into mesoscopic theories and molecular mechanisms for curvature sensing.
Cholesterol is an essential component of all animal cell membranes and plays an important role in maintaining the membrane structure and physical–chemical properties necessary for correct cell functioning. The presence of cholesterol is believed to be responsible for domain formation (lipid rafts) due to different interactions of cholesterol with saturated and unsaturated lipids. In order to get detailed atomistic insight into the behaviour of cholesterol in bilayers composed of lipids with varying degrees of unsaturation, we have carried out a series of molecular dynamics simulations of saturated and polyunsaturated lipid bilayers with different contents of cholesterol, as well as well-tempered metadynamics simulations with a single cholesterol molecule in these bilayers. From these simulations we have determined distributions of cholesterol across the bilayer, its orientational properties, free energy profiles, and specific interactions of molecular groups able to form hydrogen bonds. Both molecular dynamics and metadynamics simulations showed that the most unsaturated bilayer with 22:6 fatty acid chains shows behaviour which is most different from other lipids. In this bilayer, cholesterol is relatively often found in a "flipped" configuration with the hydroxyl group oriented towards the membrane middle plane. This bilayer has also the highest (least negative) binding free energy among liquid phase bilayers, and the lowest reorientation barrier. Furthermore, cholesterol molecules in this bilayer are often found to form head-to-tail contacts which may lead to specific clustering behaviour. Overall, our simulations support ideas that there can be a subtle interconnection between the contents of highly unsaturated fatty acids and cholesterol, deficiency or excess of each of them is related to many human afflictions and diseases.
The all-atomic force field Slipids (Stockholm Lipids) for lipid bilayers simulations has been extended to polyunsaturated lipids. Following the strategy adopted in the development of previous versions of the Slipids force field, the parametrization was essentially based on high-level ab initio calculations. Atomic charges and torsion angles related to polyunsaturated lipid tails were parametrized using structures of dienes molecules. The new parameters of the force field were validated in simulations of bilayers composed of seven polyunsaturated lipids. An overall good agreement was found with available experimental data on the areas per lipids, volumetric properties of bilayers, deuterium order parameters, and scattering form factors. Furthermore, simulations of bilayers consisting of highly polyunsaturated lipids and cholesterol molecules have been carried out. The majority of cholesterol molecules were found in a position parallel to bilayer normal with the hydroxyl group directed to the membrane surface, while a small fraction of cholesterol was found in the bilayer center parallel to the membrane plane. Furthermore, a tendency of cholesterol molecules to form chain-like clusters in polyunsaturated bilayers was qualitatively observed.
A series of 19 hydroxylated polybrominated diphenyl ethers (OH-PBDEs) have been studied using density functional theory (DFT) and molecular dynamics simulations with the purpose of investigating eventual correlations between their physicochemical properties and toxic action. Dissociation constants (pK(a)), solvation free energies and octanol-water partition coefficients (logP) have been computed. Additionally, metadynamics simulations of OH-PBDEs passing through a lipid bilayer have been carried out for four OH-PBDE species. No correlations between computed pKa values and toxicity data have been found. Medium correlations were found between partition coefficients and the ability of OH-PBDEs to alter membrane potential in cell cultures, which is attributed to higher uptake of molecules with larger log P parameters. It was also demonstrated that in lipid bilayers, OH-PBDE molecules differ in their orientational distributions and can adopt different conformations which can affect the uptake of these molecules and influence the pathways of their toxic action.
In the eukaryotic cell nucleus, DNA exists as chromatin, a compact but dynamic complex with histone proteins. The first level of DNA organization is the linear array of nucleosome core particles (NCPs). The NCP is a well-defined complex of 147 bp DNA with an octamer of histones. Interactions between NCPs are of paramount importance for higher levels of chromatin compaction. The polyelectrolyte nature of the NCP implies that nucleosome-nucleosome interactions must exhibit a great influence from both the ionic environment as well as the positively charged and highly flexible N-terminal histone tails, protruding out from the NCP. The large size of the system precludes a modelling analysis of chromatin at an all-atom level and calls for coarse-grained approximations. Here, a model of the NCP that include the globular histone core and the flexible histone tails described by one particle per each amino acid and taking into account their net charge is proposed. DNA wrapped around the histone core was approximated at the level of two base pairs represented by one bead (bases and sugar) plus four beads of charged phosphate groups. Computer simulations, using a Langevin thermostat, in a dielectric continuum with explicit monovalent (K+), divalent (Mg2+) or trivalent (Co(NH3)(6)(3+)) cations were performed for systems with one or ten NCPs. Increase of the counterion charge results in a switch from repulsive NCP-NCP interaction in the presence of K+, to partial aggregation with Mg2+ and to strong mutual attraction of all 10 NCPs in the presence of CoHex(3+). The new model reproduced experimental results and the structure of the NCP-NCP contacts is in agreement with available data. Cation screening, ion-ion correlations and tail bridging contribute to the NCP-NCP attraction and the new NCP model accounts for these interactions.
The magnetic 2D to 3D crossover behavior of well-ordered arrays of monodomain gamma-Fe2O3 spherical nanoparticles with different thicknesses has been investigated by magnetometry and Monte Carlo (MC) simulations. Using the structural information of the arrays obtained from grazing incidence small-angle X-ray scattering and scanning electron microscopy together with the experimentally determined values for the saturation magnetization and magnetic anisotropy of the nanoparticles, we show that MC simulations can reproduce the thickness-dependent magnetic behavior. The magnetic dipolar particle interactions induce a ferromagnetic coupling that increases in strength with decreasing thickness of the array. The 2D to 3D transition in the magnetic properties is mainly driven by a change in the orientation of the magnetic vortex states with increasing thickness, becoming more isotropic as the thickness of the array increases. Magnetic anisotropy prevents long-range ferromagnetic order from being established at low temperature and the nanoparticle magnetic moments instead freeze along directions defined by the distribution of easy magnetization directions.
In this work, we present results from molecular dynamics simulations of aqueous solutions of furfural and 5-hydroxymethylfurfural, which are important intermediates in the hydrothermal carbonization processes of biomass conversion. The computations were performed both at ambient and hydrothermal conditions using a two-level factorial design varying concentration, temperature, and pressure. A number of equilibrium and dynamic properties have been computed including enthalpies and free energies of vaporization, free energies of solvation, diffusion coefficients, and rotational/reorientational correlation times. Structural properties of solutions were analyzed using radial and spatial distribution functions. It was shown that the formation of hydrogen bonds among 5-hydroxymethylfurfural molecules is preferred compared to hydrogen bonding between 5-hydroxymethylfurfural and water. In addition, our results suggest that the oxygen atoms in the furan rings of furfural and 5-hydroxymethylfurfural do not participate in hydrogen bonding to the same extent as the oxygen atoms in the hydroxyl and carbonyl groups. It is also observed that furfural molecules aggregate under certain conditions, and we show how this is affected by changes in temperature, pressure, and concentration in agreement with experimental solubility data. The analysis of the computational results provides useful insight into the structure and dynamics of the considered molecules at conditions of hydrothermal carbonization, as well as at ambient conditions.
Small-angle X-ray scattering (SAXS) is used to demonstrate the presence of density fluctuations in ambient water on a physical length-scale of ≈1 nm; this is retained with decreasing temperature while the magnitude is enhanced. In contrast, the magnitude of fluctuations in a normal liquid, such as CCl4, exhibits no enhancement with decreasing temperature, as is also the case for water from molecular dynamics simulations under ambient conditions. Based on X-ray emission spectroscopy and X-ray Raman scattering data we propose that the density difference contrast in SAXS is due to fluctuations between tetrahedral-like and hydrogen-bond distorted structures related to, respectively, low and high density water. We combine our experimental observations to propose a model of water as a temperature-dependent, fluctuating equilibrium between the two types of local structures driven by incommensurate requirements for minimizing enthalpy (strong near-tetrahedral hydrogen-bonds) and maximizing entropy (nondirectional H-bonds and disorder). The present results provide experimental evidence that the extreme differences anticipated in the hydrogen-bonding environment in the deeply supercooled regime surprisingly remain in bulk water even at conditions ranging from ambient up to close to the boiling point.
The influence of local anesthetic lidocaine on electrostatic properties of a lipid membrane bilayer was studied by molecular dynamics simulations. The electrostatic dipole potential, charge densities, and orientations of the headgroup angle have been examined in presence of different amounts of charged or uncharged forms of lidocaine. Important differences of the membrane properties caused by the presence of the both forms of lidocaine are presented and discussed. Our simulations have shown that both charged and uncharged lidocaine cause almost the same increase of the dipole electrostatic potential in the middle of membrane though for different reasons. The increase, being about 90 mV for 9 mol % of lidocaine and 220 mV for 28 mol% of lidocaine, is of the size which may affect the functioning of voltage-gated ion channels.
The CHARMM force field for DMPC lipids was modified in order to improve agreement with experiment for a number of important properties of hydrated lipid bilayer. The modification consists in introduction of a scaling factor 0.83 for 1-4 electrostatic interactions (between atoms separated by three covalent bonds), which provides correct transgauche ratio in the alkane tails, and recalculation of the headgroup charges on the basis of HF/6-311(d,p) ab-initio computations. Both rigid TIP3P and flexible SPC water models were used with the new lipid model, showing similar results. The new model in a 75 ns simulation has shown a correct value of the area per lipid at zero surface tension, as well as good agreement with the experiment for the electron density, structure factor, and order parameters, including those in the headgroup part of lipids.
The Bead-Fourier path integral molecular dynamics technique introduced earlier [ S. D. Ivanov, A. P. Lyubartsev, and A. Laaksonen, Phys. Rev. E 67 066710 (2003) ] is applied for simulation of electrons in the simplest molecules: molecular hydrogen, helium atom, and their ions. Special attention is paid to the correct description of electrons in the core region of a nucleus. In an attempt to smooth the Coulomb potential at small distances, a recipe is suggested. The simulation results are in excellent agreement with the analytical solution for the "harmonic helium atom", as well as with the vibrational potential of the H2 molecule and He ionization energies. It is demonstrated, that the Bead-Fourier path integral molecular dynamics technique is able to provide the accuracy required for the description of electron structure and chemical bonds in cases when electron exchange effects need not be taken into account.
The behavior of a flexible anionic chain of 150 univalent and negatively charged beads connected by harmonic-like potential interactions with each other in the presence of equal number of positive and free counterions, observed in molecular dynamics simulations with Langevin thermostat, is described in a temperature range from 0.1 to 10.0 in reduced units. The total and Coulombic energies, radial distribution functions, radii of gyration, end-to-end distances of the chain are depicted. Our results turned out to be qualitatively similar to the results obtained earlier for a lattice polyelectrolyte model, including temperature maximum of the polyelectrolyte chain and internal phase transition which seems to occur abruptly at low temperatures for all the systems studied, judging from the shape of end-to end distance, gyration radius and energy profiles.
Liposomes are proposed as drug delivery systems and can in principle be designed so as to cohere with specific tissue types or local environments. However, little detail is known about the exact mechanisms for drug delivery and the distributions of drug molecules inside the lipid carrier. In the current work, a coarse-grained (CG) liposome model is developed, consisting of over 2500 lipids, with varying degrees of drug loading. For the drug molecule, we chose hypericin, a natural compound proposed for use in photodynamic therapy, for which a CG model was derived and benchmarked against corresponding atomistic membrane bilayer model simulations. Liposomes with 21-84 hypericin molecules were generated and subjected to 10 microsecond simulations. Distribution of the hypericins, their orientations within the lipid bilayer, and the potential of mean force for transferring a hypericin molecule from the interior aqueous droplet through the liposome bilayer are reported herein.
Biological membranes are versatile in composition and host intriguing molecular processes. In order to be able to study these systems, an accurate model Hamiltonian or force field (FF) is a necessity. Here, we report the results of our extension of earlier developed all-atomistic FF parameters for fully saturated phospholipids that complements an earlier parameter set for saturated phosphatidylcholine lipids (J. Phys. Chem. B, 2012, 116, 3164-3179). The FF, coined Slipids (Stockholm lipids), now also includes parameters for unsaturated phosphatidylcholine and phosphatidylethanolamine lipids, e.g., POPC, DOPC, SOPC, POPE, and DOPE. As the extended set of parameters is derived with the same philosophy as previously applied, the resulting FF has been developed in a fully consistent manner. The capabilities of Slipids are demonstrated by performing long simulations without applying any surface tension and using the correct isothermal-isobaric (NPT) ensemble for a range of temperatures and carefully comparing a number of properties with experimental findings. Results show that several structural properties are very well reproduced, such as scattering form factors, NMR order parameters, thicknesses, and area per lipid. Thermal dependencies of different thicknesses and area per lipid are reproduced as well Lipid diffusion is systematically slightly underestimated, whereas the normalized lipid diffusion follows the experimental trends. This is believed to be due to the lack of collective movement in the relatively small bilayer patches used Furthermore, the compatibility with amino acid FFs from the AMBER family is tested in explicit transmembrane complexes of the WALP23 peptide with DLPC and DOPC bilayers, and this shows that Slipids can be used to study more complex and biologically relevant systems.
To be able to model complex biological membranes in a more realistic manner, the force field Slipids (Stockholm lipids) has been extended to include parameters for sphingomyelin (SM), phosphatidylglycerol (PG), phosphatidylserine (PS) lipids, and cholesterol. Since the parametrization scheme was faithful to the scheme used in previous editions of Slipids, all parameters are consistent and fully compatible. The results of careful validation of a number of key structural properties for one and two component lipid bilayers are in excellent agreement with experiments. Potentials of mean force for transferring water across binary mixtures of lipids and cholesterol were also computed in order to compare water permeability rates to experiments. In agreement with experimental and simulation studies, it was found that the permeability and partitioning of water is affected by cholesterol in lipid bilayers made of saturated lipids to the largest extent. With the extensions of Slipids presented here, it is now possible to study complex systems containing many different lipids and proteins in a fully atomistic resolution in the isothermic-isobaric (NPT) ensemble, which is the proper ensemble for membrane simulations.
An all-atomistic force field (FF) has been developed for fully saturated phospholipids. The parametrization has been largely based on high-level ab initio calculations in order to keep the empirical input to a minimum. Parameters for the lipid chains have been developed based on knowledge about bulk alkane liquids, for which thermodynamic and dynamic data are excellently reproduced. The FFs ability to simulate lipid bilayers in the liquid crystalline phase in a tensionless ensemble was tested in simulations of three lipids: 1,2-diauroyl-sn-glycero-3-phospocholine (DLPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and 1,2-dipalmitoyl-sn-glycero-3-phospcholine (DPPC). Computed areas and volumes per lipid, and three different kinds of bilayer thicknesses, have been investigated. Most importantly NMR order parameters and scattering form factors agree in an excellent manner with experimental data under a range of temperatures. Further, the compatibility with the AMBER FF for biomolecules as well as the ability to simulate bilayers in gel phase was demonstrated. Overall, the FF presented here provides the important balance between the hydrophilic and hydrophobic forces present in lipid bilayers and therefore can be used for more complicated studies of realistic biological membranes with protein insertions.
Free energy calculations are vital for our understanding of biological processes on an atomistic scale and can offer insight to various mechanisms. However, in some cases, degrees of freedom (DOFs) orthogonal to the reaction coordinate have high energy barriers and/or long equilibration times, which prohibit proper sampling. Here we identify these orthogonal DOFs when studying the transfer of a solute from water to a model membrane. Important DOFs are identified in bulk liquids of different dielectric nature with metadynamics simulations and are used as reaction coordinates for the translocation process, resulting in two- and three-dimensional space of reaction coordinates. The results are in good agreement with experiments and elucidate the pitfalls of using one-dimensional reaction coordinates. The calculations performed here offer the most detailed free energy landscape of solutes embedded in lipid bilayers to date and show that free energy calculations can be used to study complex membrane translocation phenomena.
We propose an effective and straightforward way of including atomic polarization in simulations of the partitioning of small molecules in inhomogenous media based on classical molecular dynamics with non-polarizable force fields. The approach presented here takes advantage of the relatively fast sampling of phase space obtained with additive force fields by adding the polarization effects afterwards. By using pre-polarized charges for the polar and non-polar phases together with a polarization correction term the effects of atomic polarization are effectively taken into account. The results show a clear improvement compared to using the more common setup with one set of charges obtained from gas phase ab initio calculations. It is shown that when proper measures are taken into account computer simulations with non-polarizable force fields are able to accurately determine water-membrane partitioning and preferential location of small molecules in the membrane interior. We believe that the approach presented here can be useful in rational drug design and in investigations of molecular mechanisms of anesthetic or toxic action.
An approach to a straightforward reparametrization of the general AMBER force field (GAFF) for small organic solutes and druglike compounds is presented. The parametrization is based on specific pair interactions between the solvent and the solute, namely, the interactions between the constituting atoms of the solute and the oxygen of water were tuned in order to reproduce experimental hydration free energies for small model compounds. The key of the parametrization was to abandon the Lorentz-Berthelot combination rules for the van der Waals interactions. These parameters were then used for larger solutes in order to validate the newly derived pair interactions. In total close to 600 hydration free energies are computed, ranging from simple alkanes to multifunctional drug compounds, and compared to experimental data. The results show that the proposed parameters work well in describing the interactions between the solute and the solvent and that the agreement in absolute numbers is good. This modified version of GAFF is a good candidate for computing and predicting hydration free energies on a large scale, which has been a long-sought goal of computational chemists and can be used in rational drug design.
Free energies of solvation (Delta G) in water and n-octanol have been computed for common drug molecules by molecular dynamics simulations with an additive fixed-charge force field. The impact of the electrostatic interactions was investigated by computing the partial atomic charges with four methods that all fit the charges from the quantum mechanically determined electrostatic potential (ESP). Due to the redistribution of electron density that occurs when molecules are transferred from gas phase to condensed phase, the polarization impact was also investigated. By computing the partial atomic charges with the solutes placed in a conductor-like continuum, the charges were effectively polarized to take the polarization effects into account. No polarization correction term or similar was considered, only the partial atomic charges. Results show that free energies are very sensitive to the choice of atomic charges and that Delta G can differ by several k(B)T depending on the charge computing method. Inclusion of polarization effects makes the solutes too hydrophilic with most methods and in vacuo charges make the solutes too hydrophobic. The restrained-ESP methods together with effectively polarized charges perform well in our test set and also when applied to a larger set of molecules. The effect of water models is also highlighted and shows that the conclusions drawn are valid for different three-point models. Partitioning between an aqueous and a hydrophobic phase is also described better if the two environment's polarization is taken into account, but again the results are sensitive to the charge calculation method. Overall, the results presented here show that effectively polarized charges can improve the description of solvating a drug-like molecule in a solvent and that the choice of partial atomic charges is crucial to ensure that molecular simulations produce reliable results.
Accurate estimation of protein-carbohydrate binding energies using computational methods is a challenging task. Here we report the use of expanded ensemble molecular dynamics (EEMD) simulation with double decoupling for estimation of binding energies of hevein, a plant lectin with its monosaccharide and disaccharide ligands GlcNAc and (GlcNAc)(2), respectively. In addition to the binding energies, enthalpy and entropy components of the binding energy are also calculated. The estimated binding energies for the hevein-carbohydrate interactions are within the range of +/- 0.5 kcal of the previously reported experimental binding data. For comparison, binding energies were also estimated using thermodynamic integration, molecular dynamics end point calculations (MM/GBSA) and the expanded ensemble methodology is seen to be more accurate. To our knowledge, the method of EEMD simulations has not been previously reported for estimating biomolecular binding energies.
The physical mechanism of the folding and unfolding of chromatin is fundamentally related to transcription but is incompletely characterized and not fully understood. We experimentally and theoretically studied chromatin compaction by investigating the salt-mediated folding of an array made of 12 positioning nucleosomes with 177 bp repeat length. Sedimentation velocity measurements were performed to monitor the folding provoked by addition of cations Na+, K+, Mg2+, Ca2+, spermidine3+, Co(NH3)63+, and spermine4+. We found typical polyelectrolyte behavior, with the critical concentration of cation needed to bring about maximal folding covering a range of almost five orders of magnitude (from 2 μM for spermine4+ to 100 mM for Na+). A coarse-grained model of the nucleosome array based on a continuum dielectric description and including the explicit presence of mobile ions and charged flexible histone tails was used in computer simulations to investigate the cation-mediated compaction. The results of the simulations with explicit ions are in general agreement with the experimental data, whereas simple Debye-Hückel models are intrinsically incapable of describing chromatin array folding by multivalent cations. We conclude that the theoretical description of the salt-induced chromatin folding must incorporate explicit mobile ions that include ion correlation and ion competition effects.
The paper reviews our current studies on the experimentally induced cation compaction and aggregation in solutions of DNA and nucleosome core particles and the theoretical modelling of these processes using coarse-grained continuum models with explicit mobile ions and with all-atom molecular dynamics (MD) simulations. Recent experimental results on DNA condensation by cationic oligopeptides and the effects of added salt are presented. The results of MD simulations modelling DNA–DNA attraction due to the presence of multivalent ions including the polyamine spermidine and fragments of histone tails, which exhibit bridging between adjacent DNA molecules, are discussed. Experimental data on NCP aggregation, using recombinantly prepared systems are summarized. Literature data and our results of studying of the NCP solutions are compared with predictions of coarse-grained MD simulations, including the important ion correlation as well as bridging mechanisms. The importance of the results to chromatin folding and aggregation is discussed.
Double helical DNA is a negatively charged polyelectrolyte and exists in the nucleus of living cells as chromatin, a highly compacted but dynamic complex with histone proteins. The first level of DNA compaction is the linear array of the nucleosome core particles (NCP), which is a well-defined structure of 145-147 bp DNA with the histone octamer, connected by linker DNA. Higher levels of chromatin compaction include two routes which may overlap: intramolecular folding of the nucleosome array resulting in formation of the 30 nm fibre and intermolecular aggregation (self-association) between different arrays (or distant fibres of the same chromosome). This review describes how the polyelectrolyte properties of chromatin are illustrated by experimental results of folding and self-association of well-defined model chromatin, in the form of recombinant nucleosome arrays, and how these properties can be understood from computer modelling. Chromatin compaction shows considerable similarities to DNA condensation. However, the structure of condensed chromatin is sensitive to the detailed molecular features of the nucleosome-nucleosome interactions which include the influence of the histone tails and their modifications.
The packaging of genomic DNA into chromatin in the eukaryotic cell nucleus demands extensive compaction. This requires attractive nucleosome-nucleosome interactions to overcome repulsion between the negatively charged DNA segments as well as other constraints. At the same time, DNA must be dynamically accessible to the cellular machinery that operates on it. Recent progress in the experimental characterisation of the higher order structure and dynamics of well-defined chromatin fibres has stimulated the attempts at theoretical description of chromatin and the nucleosome. Here we review the present status of chromatin Modelling, with particular emphasis on coarse-grained computer simulation models, the role of electrostatic interactions, and discuss future perspectives in the field.
Computer modeling of very large biomolecular systems, such as long DNA polyelectrolytes or protein-DNA complex-like chromatin cannot reach all-atom resolution in a foreseeable future and this necessitates the development of coarse-grained (CG) approximations. DNA is both highly charged and mechanically rigid semi-flexible polymer and adequate DNA modeling requires a correct description of both its structural stiffness and salt-dependent electrostatic forces. Here, we present a novel CG model of DNA that approximates the DNA polymer as a chain of 5-bead units. Each unit represents two DNA base pairs with one central bead for bases and pentose moieties and four others for phosphate groups. Charges, intra-and inter-molecular force field potentials for the CG DNA model were calculated using the inverse Monte Carlo method from all atom molecular dynamic (MD) simulations of 22 bp DNA oligonucleotides. The CG model was tested by performing dielectric continuum Langevin MD simulations of a 200 bp double helix DNA in solutions of monovalent salt with explicit ions. Excellent agreement with experimental data was obtained for the dependence of the DNA persistent length on salt concentration in the range 0.1-100 mM. The new CG DNA model is suitable for modeling various biomolecular systems with adequate description of electrostatic and mechanical properties.
Chromatin condensation is driven by the energetically favourable interaction between nucleosome core particles (NCPs). The close NCP-NCP contact, stacking, is a primary structural element of all condensed states of chromatin in vitro and in vivo. However, the molecular structure of stacked nucleosomes as well as the nature of the interactions involved in its formation have not yet been systematically studied. Here we undertake an investigation of both the structural and physico-chemical features of NCP structure and the NCP-NCP stacking. We introduce an NCP-centred set of parameters (NCP-NCP distance, shift, rise, tilt, and others) that allows numerical characterisation of the mutual positions of the NCPs in the stacking and in any other structures formed by the NCP. NCP stacking in more than 140 published NCP crystal structures were analysed. In addition, coarse grained (CG) MD simulations modelling NCP condensation was carried out. The CG model takes into account details of the nucleosome structure and adequately describes the long range electrostatic forces as well as excluded volume effects acting in chromatin. The CG simulations showed good agreement with experimental data and revealed the importance of the H2A and H4 N-terminal tail bridging and screening as well as tail-tail correlations in the stacked nucleosomes.
To model large biomolecular systems, such as cell and organelles an atomistic description is not currently achievable and is not generally practical. Therefore, simplified coarse-grained (CG) modelling becomes a necessity. One of the most important cellular components is chromatin, a large DNA-protein complex where DNA is highly compacted. Recent progress in coarse graining modelling of the major chromatin components, double helical DNA and the nucleosome core particle (NCP) is presented. First, general principles and approaches allowing rigorous bottom-to-top generation of interaction potentials in the CG models are presented. Then, recent CG models of DNA are reviewed and their adequacy is benchmarked against experimental data on the salt dependence of DNA flexibility (persistence length). Furthermore, a few recent CG models of the NCP are described and their application for studying salt-dependent NCP-NCP interaction is discussed. An example of a multiscale approach to CG modelling of chromatin is presented where interactions and self-assembly of thousands of NCPs in solution are observed.
The positively charged N-terminal histone tails play a crucial role in chromatin compaction and are important modulators of DNA transcription, recombination, and repair. The detailed mechanism of the interaction of histone tails with DNA remains elusive. To model the unspecific interaction of histone tails with DNA, all-atom molecular dynamics (MD) simulations were carried out for systems of four DNA 22-mers in the presence of 20 or 16 short fragments of the H4 histone tail (variations of the 16-23 a. a. KRHRKVLR sequence, as well as the unmodified fragment a. a. 13-20, GGAKRHRK). This setup with high DNA concentration, explicit presence of DNA-DNA contacts, presence of unstructured cationic peptides (histone tails) and K+ mimics the conditions of eukaryotic chromatin. A detailed account of the DNA interactions with the histone tail fragments, K+ and water is presented. Furthermore, DNA structure and dynamics and its interplay with the histone tail fragments binding are analysed. The charged side chains of the lysines and arginines play major roles in the tailmediated DNA-DNA attraction by forming bridges and by coordinating to the phosphate groups and to the electronegative sites in the minor groove. Binding of all species to DNA is dynamic. The structure of the unmodified fully-charged H4 16-23 a. a. fragment KRHRKVLR is dominated by a stretched conformation. The H4 tail a. a. fragment GGAKRHRK as well as the H4 Lys16 acetylated fragment are highly flexible. The present work allows capturing typical features of the histone tail-counterionDNA structure, interaction and dynamics.
We review methods to compute x-ray absorption spectra (XAS) with special focus on the transition potential approach of Triguero et al. [Phys. Rev. B 58, 8097 (1998)] and its application to calculations on water in condensed phase. We discuss the absolute energy scale, functional dependence, broadening versus sampling of intra- and intermolecular vibrational modes, treatment of the continuum, cluster size convergence as well as compare with periodic calculations and with experiment; periodic and cluster model calculations are found to agree very closely in the relevant near-edge region although neither reproduces the pre-edge and main-edge features in the experimental spectra of thin ice films. The real space grid representation of the wave function in the periodic calculations allows a more extended energy range to be described and we find satisfactory agreement with experiment for higher energy continuum resonances. Two proposed alternative approaches using either the potential from a full core-hole (FCH) or the full core-hole with an excited electron in the lowest state (XCH) are shown to lead to spectra that deviate significantly from experiment.
We analyze the validity of the commonly used electric-field (E-field) approximation to vibrational OH stretch Raman spectra of dilute HOD in D2O by computing the OH stretch frequency of all molecules in several different structure models, each containing around 2000 molecules. The calculations are done at the B3LYP level using clusters containing 32 molecules centered around the molecule for which the frequencies are calculated; the large cluster size is required due to significant nonlocal contributions influencing the computed frequencies. The vibrational frequencies are determined using a six-point potential optimized discrete variable representation. Raman and infrared intensities are furthermore computed to generate the spectra. We find that a quadratic fit of E-field versus frequency gives a reasonable representation of the calculated distribution of frequencies. However, the mapping depends significantly on the structural model and is thus not universal. Anharmonic couplings are calculated for several optimized clusters showing a general trend to compress the computed frequency distributions, which is in agreement with dynamical simulations (motional narrowing).
Wehave carried out Monte Carlo simulation on the primitive onedimensional model for water described earlier [A. Ben-Naim, J. Chem.Phys. 128, 024506 (2008)]. We show that by taking intoaccount second nearest neighbor interactions, one can obtain the characteristicanomalous solvation thermodynamic quantities of inert solutes in water. Thismodel clearly demonstrates the molecular origin of the large negativeentropy of solvation of an inert solute in water.
Systematic construction of coarse-grained molecular models from detailed atomistic simulations, and even from ab initio simulations is discussed. Atomistic simulations are first performed to extract structural information about the system, which is then used to determine effective potentials for a coarse-grained model of the same system. The statistical-mechanical equations expressing the canonical properties in terms of potential parameters can be inverted and solved numerically according to the iterative Newton scheme. In our previous applications, known as the Inverse Monte Carlo, radial distribution functions were inverted to reconstruct pair potential, while in a more general approach the targets can be other canonical averages. We have considered several examples of coarse-graining; for the united atom water model we suggest an easy way to overcome the known problem of high pressure. Further, we have developed coarse-grained models for L- and D-prolines, dissolved here in an organic solvent (dimethylsulfoxide), keeping their enantiomeric properties from the corresponding all-atom proline model. Finally, we have revisited the previously developed coarse-grained lipid model based on an updated all-atomic force field. We use this model in large-scale meso-scale simulations demonstrating spontaneous formation of different structures, such as vesicles, micelles, and multi-lamellar structures, depending on thermodynamical conditions.
The nucleosome core particle (NCP) is the basic building block of chromatin. Under the influence of multivalent cations, isolated mononucleosomes exhibit a rich phase behaviour forming various columnar phases with characteristic NCP-NCP stacking. NCP stacking is also a regular element of chromatin structure in vivo. Understanding the mechanism of nucleosome stacking and the conditions leading to self-assembly of NCPs is still incomplete. Due to the complexity of the system and the need to describe electrostatics properly by including the explicit mobile ions, novel modelling approaches based on coarse-grained (CG) methods at the multiscale level becomes a necessity. In this work we present a multiscale CG computer simulation approach to modelling interactions and self-assembly of solutions of NCPs induced by the presence of multivalent cations. Starting from continuum simulations including explicit three-valent cobalt(III) hexammine (CoHex(3+)) counterions and 20 NCPs, based on a previously developed advanced CG NCP model with one bead per amino acid and five beads per two DNA base pair unit (Fan et al 2013 PLoS One 8 e54228), we use the inverse Monte Carlo method to calculate effective interaction potentials for a 'super-CG' NCP model consisting of seven beads for each NCP. These interaction potentials are used in large-scale simulations of up to 5000 NCPs, modelling self-assembly induced by CoHex(3+). The systems of 'super-CG' NCPs form a single large cluster of stacked NCPs without long-range order in agreement with experimental data for NCPs precipitated by the three-valent polyamine, spermidine(3+).
We outline our coarse-graining strategy for linking micro-and mesoscales of soft matter and biological systems. The method is based on effective pairwise interaction potentials obtained in detailed ab initio or classical atomistic Molecular Dynamics (MD) simulations, which can be used in simulations at less accurate level after scaling up the size. The effective potentials are obtained by applying the inverse Monte Carlo (IMC) method [A. P. Lyubartsev and A. Laaksonen, Phys. Rev. E 52(4), 3730-3737 (1995)] on a chosen subset of degrees of freedom described in terms of radial distribution functions. An in-house software package MagiC is developed to obtain the effective potentials for arbitrary molecular systems. In this work we compute effective potentials to model DNA-protein interactions (bacterial LiaR regulator bound to a 26 base pairs DNA fragment) at physiological salt concentration at a coarse-grained (CG) level. Normally the IMC CG pair-potentials are used directly as look-up tables but here we have fitted them to five Gaussians and a repulsive wall. Results show stable association between DNA and the model protein as well as similar position fluctuation profile.
With the rapid development of computer power and wide availability of modelling software computer simulations of realistic models of lipid membranes, including their interactions with various molecular species, polypeptides and membrane proteins have become feasible for many research groups. The crucial issue of the reliability of such simulations is the quality of the force field, and many efforts, especially in the latest several years, have been devoted to parametrization and optimization of the force fields for biomembrane modelling. In this review, we give account of the recent development in this area, covering different classes of force fields, principles of the force field parametrization, comparison of the force fields, and their experimental validation. This article is part of a Special Issue entitled: Biosimulations edited by Ilpo Vattulainen and Tomasz Rog.


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From Cristiano Ronaldo to Ronaldinho or even Joey Barton, Twitter is full of profiles – real and fake – of football players with an interesting story.
But with all the abundance of information travelling through the world-wide-web it is very hard to keep track of everything. In this short post, we make a quick overview of the top list of the most followed and most interesting (even a bit bizarre) accounts out there.
Most followed
Although Kaka (@KAKA) was the first ever athlete to gain more than 10 million followers on Twitter, it is his team mate Cristiano Ronaldo (@Cristiano) who has the most followers at the moment – over 18 million. Kaka, even though he is far away from his best days at AC Milan, became a social media celebrity by actively engaging with fans – especially in the beginning, he was good at replying to their messages –, while Ronaldo's account looks like a well planned marketing tool from the start. Writing only in English (Kaka also uses his mother tongue), he is easier to follow for most fans around the world and doesn't let the world too much into his private life. He – like most top players – probably does not write his own tweets. Both mentioned players also have a lot of "fake" followers – according to Mashable around 40%! It should be noted that most people on Twitter have a significant share of "fake" followers: profiles made online with whatever reason that does not exist in the real world.
Are you a footballler and want to take the next step in your career?
Most controversial
Joey Barton (@Joey7Barton) is surely a gifted football player, but far away from being one of the best out there. But if there is one thing he is better than a vast majority of professionals around the world, it is controversy. He has got a lot of points to prove not only on the field but also on Twitter. Sadly for him, he often posts tweets and says things he later regrets. Later on, he may try to compensate with apologies or even with promoted tweets – like he did after receiving a red card while still playing in England; he also just recently described the late British PM Margaret Thatcher an "old witch" and polarized the public once again. With over two million followers, it is clearly visible that there are also people who value his virtue, sharing genuine thoughts of a well-known athlete with the world.
One of the best parodies
While it is not even somebody from his PR team writing tweets for him, Mario Balotelli has a strong presence on Twitter anyway. The platform is full of his parodies, made as a tribute to the talented, but troublesome player. @MarioBaloltelli is one of the best parody accounts on Twitter. It has more than half a million followers as the person running the account keeps posting funny posts, with "Balotelli-like" comments, such as the rumour that Gonzalo Higuain was born offside, or his famous good night wish from a few weeks ago: "Night night, sleep tight, don't let the Suarez bite."
Still loved
It has been years, since he has played in Europe and done magic with the ball, but people have not forgotten him. Although Ronaldinho (@10Ronaldinho) is now playing in Brazil, where he continues to produce goals only he is capable of, and only tweets in Portuguese, he has one of the most accessed accounts on the world with almost 6 million followers. Even if you don't understand a bit what he writes, the "behind-the-scenes" photos give excellent added value. At the age of 33 it is obvious that he is still not only a very charismatic, recognized and loved footballer, but he can still score wonderful goals.
What about clubs?
Except for fans, clubs are not as fun to follower as individual players, as they (obviously) do not really have the most important thing to be attractive on Twitter – a personality. Most clubs are very careful and use Twitter strictly as a PR tool, but are not equally successful even at that. While Spanish outfits FC Barcelona (@FCBarcelona) and Real Madrid (@realmadrid) predictably lead the chart, there are also two very successful Turkish clubs on Twitter – Galatasaray (@GalatasaraySK) and Fenerbahce (@Fenerbahce) are 3rd and 5th on the chart. Right among them is Arsenal FC (@ArsenalFC) who has a very important and innovative role when it comes to the web – and not only on social media. Unlike some other clubs, they know what the fans want and they deliver it to them.
Ready to kickstart your career as football agent?
Related articles
- 5 tips for footballers: How not to use Twitter
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- Fieldoo launches agents' platform – a unique football marketplace!
- Joey Barton banned for two games for Thiago Silva overweight ladyboy tweets
- There was nothing like Fieldoo when I was a young player – but I wish there was (by @FourFourTom)
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- 10 useful tips for better use of social networks (part 2)

subset: commoncrawl


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Kayaking the Kiamichi River
Originally uploaded by FreeWine
Any Oklahoman with a digital camera and an interest in the outdoors is eligible to see their work published in this years Showcase edition of Outdoor Oklahoma magazine.
Submissions are being accepted through March 31, and winners will have their work featured in the July/August 2008 issue.
Although the editors of Outdoor Oklahoma encourage readers to submit images including a variety of outdoor-related subjects, the magazine has been focusing on "faces in the outdoors" to show hunters, anglers, kids and other outdoor enthusiasts enjoying the outdoors.
Each participant may submit up to five digital images. Each submission must include a description of the photo, including the location taken, names and hometowns of subjects and what it took to get just the right shot. Photos should be in sharp focus, and images should be at least 300 dpi (dots per inch). The canvas size should be about 8 inches by 11 inches. Slides and print images will not be accepted.
Hopeful photographers can mail a disk to:
"Outdoor Oklahoma" Magazine
Oklahoma Dept. of Wildlife Conservation
P.O. Box 53465
Oklahoma City, OK 73152.
Readers may also e-mail their entries to email@example.com.
Photography Oklahoma Outdoors Contest


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Simple & Secure Ordering!
Regular price
12" Flower Pot Holder with Special screw mounting brackets clasp to VERTICAL wood balusters. Great for apartment home railings, decks, fences and porches. Fits 1" to 1-1/2" square balusters / spires. The brackets are made with specially formed 1/8" thick steel plate which provides superior holding capability. Very simple and easy to install with only a screw driver.Powder coated 7/16" square steel.


text: The one and only one experimental hero of Tollywood Nagarjuna facing series of flops from past years. Nagarjuna last profitable venture was King and then Gaganam, secured minimal profits for the producer.After King Nagarjuna's Kedi remained as a disaster , and then Ragada did very decent for distributors with break even numbers. Bi-lingual movie Gaganam got minimal profits and then Rajanna ,got very high positive talk and ended at single digit share .Shirid Sai also failed at box-office. Dhamarukam which is the biggest earner in Nag's career ended as commercial flop due to high budget and financial issues. Coming to his recent venture Greekuveerudu made with moderate budget and producer sold for 15C . Movie opened with mixed talk but dropped heavily after first weekend. Movie came to close in third week it self. In Krishna district Greekuveerudu collected less share than disaster Shadow. Almost all the distributors may loose amount on Greekuveerudu. This is not a situation of Nagrjuna , it applies for Senior heroes like Balakrishna and Venkatesh.


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This week's Arrow had some great moments and some plot holes. I'm really enjoying the series so far but it is basically playing for me like a Batman series without the name Batman. Rich playboy puts on a facade, is super serious in his underground lair, and now his manservant/companion is his confidant? Pffffff. I mean, seriously. Maybe I don't know enough about Oliver/Green Arrow to tell the difference, but what's the difference?
Discussion points/thoughts copied over from tweets:
1) Somehow I missed that Thea is supposed to be in high school. She looks like a college brat if anything. She's still one of the best actresses on the show, but I have trouble believing high school. Ah well.
2) OK, I just want more Oliver jumping around on buildings and climbing things, may I have more of that please? Because that is hot shit.
3) So how many languages does Ollie know? I mean, I got that he spoke mandarin or whatever because of the island, but when the hell did he learn Russian? I fear the inevitable episode when he butchers Japanese. But I fear all episodes where people butcher Japanese. (thank you Supernatural and Heroes for doing it right most of the time)
4) I still love Tommy Merlin [sp?] more than life itself. He is a hottie pants. His bromance with Ollie is totally shippable. And GOD, he looks good in leather. I know they're just doing the Tommy/
5) I have a great affinity for identity reveals. The part where the hero takes off his mask is invariably one of my favorite moments of almost any media. So I was jumping up and down when this one happened. I really look forward to seeing Ollie trust someone. And he couldn't pick a better someone to trust, either. I look forward to seeing Diggs choose his alliances. Ollie's not the one paying him, after all.
6) OH HALLO REFERENCE TO BLUDHAVEN IN THE PREVIEW. :D (can we please have nightwing please please please please how bout matt cohen)
That's that, have at it! SPN meta coming next.


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Drugs, allies, and enemies: CNN Philippines goes one-on-one with President Duterte
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Metro Manila (CNN Philippines) — President Rodrigo Duterte sat down with CNN Philippines for his first-ever live interview with a local media organization since he took office.
From the collateral damage of the drug war, to the few alliances he holds close, here's a wrap of all the big issues Duterte opened up about.
Close allies
Duterte said he didn't fear or favor anyone in his crackdown on illegal drugs. He said other politicians would exert influence to have police call off anti-drug operations when these began affecting their allies and supporters. In contrast, the President said he owes no debt of gratitude ("utang na loob") to anyone.
Except, however, to "three or four persons."
The President named Ilocos Norte Governor Imee Marcos as the first one, saying he promised her he would allow the burial of her father, the late President Ferdinand Marcos, in the Libingan ng mga Bayani.
Duterte also named Bukidnon Governor Jose Ma. Zubiri, Jr. — the father of Senator Juan Miguel Zubiri — as well as Bataan Governor Abet Garcia.
Cabinet changes
Senator Alan Peter Cayetano, Duterte's running mate, is in line to become the next Foreign Affairs Secretary. The current chief, Perfecto Yasay, will be returning to the United States, where he teaches law at the University of Hawaii.
There is no post yet, however, for former Senator Bongbong Marcos, Imee's brother. "I do not want to make promises," the President said.
Duterte said the younger Marcos had wanted to head the Department of Tourism, but election rules require him to sit out of government for a year after he ran and lost the vice-presidential race in May.
Robredo split
Duterte is "not considering" a new Cabinet post for Vice-President Leni Robredo after she resigned earlier this month.
The President said he held "nothing against Leni," but he pointed out it was "incongruous" of her to to be part of his team when her very own party wanted to oust him.
Robredo had chaired the Housing and Urban Development Coordinating Council, but she stepped down when Duterte barred her from Cabinet meetings.
Ouster plots
According to Duterte, the United States and the Liberal Party are behind the plots to oust him.
He challenged them to do their best and said he would fight back and "give them a bloody nose."
Duterte added he didn't need the presidency to begin with. He said he only wants to stay to "honor [his] contract" with those who voted him to power.
U.S. military ties
The President wants to abolish the Visiting Forces Agreement (VFA) between the Philippines and the United States. He said: "Why won't I kick them out? They want to destroy me. Who will be a stupid person, a President that will allow that?"
He admitted, though, that he needs to review the consequences of scrapping the treaty. Many oppose his plans, including his own military leaders.
The Philippines will assert its rights if China tries to "siphon minerals" from the South China Sea. While the government is open to negotiations, even joint exploration in the disputed waters, the President said he would raise the arbitral decision if China exploited resources in the disputed waters.
"I have this title," Duterte said of the international court ruling. "We share economic bounties. So how about us?"
Drug war
Duterte recognized some innocent lives have been lost in the administration's war on drugs. But he stressed, these were only victims caught in the crossfire. Everyone else targeted by police operations were "not suspects, but criminals."
The President also warned the government's war on drugs would continue all throughout his term. He said he wouldn't stop "until the last drug lord is killed."
Watch the livestream of CNN Philippines' One-on-One with the President here.
Catch the replays of the interview on December 30 (10 AM and 11 PM), December 31 (9 AM and 8 PM), and January 1 (10 AM) on CNN Philippines.


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Even its authors would be unlikely to expect the title of the 24-page document to set the pulse racing. But the conclusions of the "Long Term Rolling Stock Strategy for the Rail Industry" - published to little fanfare last month - are startling and present the current Government with challenges radically different from those faced by the Conservative administration in its death throes during the early 1960s.
Written by Network Rail, the Association of Train Operators and major rolling stock companies, it sets out the industry's forecast for the next 30 years and gets over one very simple message - the rail industry has failed to keep up with the surge in demand for train travel.
"Passenger miles" - the total number of miles travelled by all passengers - grew by 91pc between 1996 and 2012. But the rail fleet increased by only 12pc over the same period.
Some cities have seen an astonishing growth. Coventry, for example, has seen journeys increase by 30 per cent in just five years, while Plymouth has reported a 38 per cent rise in traffic over the same period
The industry now accepts this is not tenable and substantial investment is needed to produce a network which is fit for purpose. This means more lines whether it be the 351-mile high speed rail network, the resurrection of those cut by Beeching or completely new schemes like Crossrail.
Significantly, the foreword to the thick document is written by Simon Burns, the rail minister, which indicates the industry's case is seen as persuasive within Whitehall.
Unlike their 1960s counterparts, who believed the spread of car ownership spelled the end of the rail industry, today's transport ministers accept huge investment is needed to keep up with demand.
"At the time the Government thought it was the right approach, but the world is a very different place now," Mr Burns told The Daily Telegraph.
Much of the change is due to the privatisation of the industry in the mid 1990s, he said. "Rail companies respond much more sensitively to market need and under successive Governments you have seen significant investment. Under the old structure if a government ran into financial difficulties, one of the easy ways to cut spending was on the railways."
The reverse is now happening. Already three lines which disappeared when Dr Beeching swung his axe have re-opened. Trains are once more running on the Chase Line between Birmingham and Walsall, on the Robin Hood Line linking Nottingham, Mansfield and Worksop as well as the Ebbw Valley Line from Ebbw Vale to Cardiff.
Plans are in place for the re-opening of the Varsity Line between Oxford and Cambridge, with the first phase from Oxford to Milton Keynes and Bedford already agreed, while work is already under way on the Waverley route from Edinburgh to Galashiels.
Stations which were left to grow tumbleweed have been restored. Chandlers Ford in Hampshire, which shut in 1969, reopened 10 years ago and now used by 100,000 passengers a year.
Demand is growing for the resurrection of a raft of other lines including one running from Uckfield to Lewes line in East Sussex.
This would, of course, provide additional services for voters in the constituency of another transport minister, Norman Baker, although he will be precluded from taking any part in the decision on whether the line should be re-opened.
The Beeching report - "The Reshaping of British Railways" - had a devastating impact on the country.
More than 2,000 stations were shut down amid a torrent of protest from communities who lost their rail services. Several thousands miles of line were rendered redundant - though exactly how much is a matter of debate - estimates vary between 4,000 to 9,000 miles.
The preface to the Beeching report observed: "First, the industry must be of a size and pattern suited to modern conditions and prospects. In particular, the railway system must be modelled to meet current needs, and the modernisation plan must be adapted to this new shape."
The same words which were used to justify the dismantling of the rail industry in the 1960s could just as easily be used now to back the dramatic expansion which all political parties believe is now essential.
Now people are using the trains in ever increasing numbers massive investment is needed, with £37.5 billion earmarked for the network between 2014-19.
The country needs carriages and engines to run on these lines, which inevitably should be good news for companies like Bombardier, who can hope to get at least some of the business.
Looking at the electric fleet alone - and a raft of lines are being converted from diesel - Britain will need 19,000 engines and carriages compared with 8,500 today.
Put another way up to 12 will have to be delivered every week if this target is to be met.
So was Beeching wrong? Richard Brown, who chaired the group which drew up the rolling stock report, believes that in hindsight he was.
"It was a different era then. " he says."Railways were the transport of the past rather than the future. Rebirth is rather a nice word now. Railways are seen as part of the solution, in the Beeching era they were part of the problem.
"Across the political spectrum there is now an acceptance that we heed to maintain people's mobility and there has been a sea change in people's point of view."
Ironically given that it was the rise of the car which spelled the death knell for the rail industry under Beeching, it is the soaring cost of motoring that has played a key part in its renaissance in recent years.
"I think the economics has shifted," Mr Brown adds. "Kids are leaving university with significant debt now. Many are working in cities and relying on public transport. Car ownership is not as aspirational as it was 50 years ago."


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Wet Cooking Sauces - includes all pasta sauces and other wet sauces in which meat and/or vegetables are cooked in (also called "pour over" sauces).
Wet Cooking Sauces (Seasonings, Dressings & Sauces) Market in Greece - Outlook to 2021: Market Size, Growth and Forecast Analytics is a broad level market review of Wet Cooking Sauces market of Greece. The research handbook provides up-to-date market size data for period 2011-2016 and illustrative forecast to 2021 covering key market aspects like Sales Value and Volume for Wet Cooking Sauces and its variants Indian Sauces, Marinades, Mexican Sauces, Oriental Sauces, Pasta Sauces and Other Wet Cooking Sauces. The research handbook also provides analytics on Sales by Brands and by Distribution Channel.
Sales Values in the handbook are depicted in USD ($) and local currency of Greece and Volumes are represented in M Kilograms.
Furthermore, the research handbook details out Sales Value for top brands for the year 2013 to 2016 and overall market sales by Distribution Channel (Hypermarkets & Supermarkets, Convenience Stores, Department Stores, Dollar Stores, Variety Store, Cash & Carries and Warehouse clubs, eRetailers, Food & Drinks specialists, Drug stores & Pharmacies, Health & Beauty Stores, Other general retailers and others) where ever applicable.
The research handbook acts as an essential tool for companies active or planning to venture in to Greece's Wet Cooking Sauces (Seasonings, Dressings & Sauces) market. The comprehensive statistics within the research handbook provides insight into the operating environment of the market and also ensures right business decision making based on historical trends and industry model based forecasting.
*Note: Certain content / sections in the research handbook may be removed or altered based on the availability and relevance of data.
- Overall Wet Cooking Sauces (Seasonings, Dressings & Sauces) market value and volume analytics with growth analysis from 2011 to 2021.
- Sales Value and Volume analytics for variants of Wet Cooking Sauces; Indian Sauces, Marinades, Mexican Sauces, Oriental Sauces, Pasta Sauces and Other Wet Cooking Sauces
- Value terms for the top brands.
- Distribution channel sales analytics from 2013-2016.
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Juan O'Sullivan's Product Testimonials
"Fantastic salsa -- the light texture and distinctive flavors that come out are always remarkable to me. I bring some with me from Des Moines to Minneapolis every time I get a chance."
– Joe Mignano - Juan's Medium Gourmet Salsa - 2 pk
"I just tasted this salsa for the first time at my parent's house. My Mom served this salsa with some tortilla chips as an early Thanksgiving snack. It's delicious!!! It's so good that I had to Google it immediately. It has a great texture and the perfect flavor! I don't know how to describe why it's SO GOOD.. but it IS!!! Great job, Juan O'Sullivan's!! Your salsa is amazing!"
– Natalie Downer - Juan's Medium Gourmet Salsa - 2 pk
" Hello Sean & Becki! I met you for the first time at last Sat. Farmers Market. I'm the one who had to try your salsa a) because I'd heard so much about it and b) because I loved the name! I bought a qt of the med salsa and as I told you after sampling, it is without a doubt the freshest, best tasting salsa I have ever had! I'm addicted to it (thanks, I hope you're happy: ) and eat it almost everyday. I'm ordering a case now online because my very good friend is having a birthday before I can get back to Market and I know she's going to love it as much as I do! Thanks so much for an awesome, homemade, Iowa product. Look forward to seeing you at Market soon and sampling the Salsa de Mesa sauce."
– Mary Rink - Juan's Medium Gourmet Salsa - 6 pk
"We first sampled Juan's medium and hot salsa at the Des Moines Home and Garden Show in February. We bought a few jars and as soon as they were gone, we ordered 12 more from the website. How convenient! Juan's medium is just the right "temperature" and texture for our tastes. We eat a lot of salsa almost daily on chips and in recipes and we like to try new brands but we have found our favorite. Our advice: order a dozen jars at a time so you won't run out."
– Troy and Jill Gullett - Juan's Medium Gourmet Salsa - 12 pk
"Juan O'Sullivan's Salsa is the best I have ever tasted! It even beats that served in restaurants! The texture is perfect and the heat is just right. Many of my friends and family - from Iowa to Arizona agree! I also love supporting entrepreneurs here in Iowa! Thank you Sean and Becky for a wonderful product!"
– Laura Pearson - Juan's Medium Gourmet Salsa - 2 pk
"Juan's chili garden, salsa, and sauces are a homemade culinary delight. Don't forget to try some of the recipes. I have had the Enchiladas By Becki Sullivan - A.K.A. - JuanÃta O'Sullivan. Very nice!!"
– Neil Feldhacker - Juan's Medium Gourmet Salsa - 2 pk
"Juan's Salsa is perfect! It is flavorful and has a great kick without being overwhelming! Other brands cannot compete with the Juan's texture and flavor. You get what you paid for Salsa! Not some vegetables with a super watery base. I have started adding Juan's to things I would never considered suitable for Salsa. My fiancé cannot stop eating it! So much, in fact, I have nicknamed their Hot Salsa, "The Rick Dawson.""
– Alexi Jameson - Juan's Medium Gourmet Salsa - 12 pk
"This salsa is AWESOME!! I first tried it a couple years ago, purchasing it at the DSM Downtown Farmer's Market....and I was HOOKED! It makes every other salsa I have tried pale by comparison. Keep up your excellent work!!!!!"
– Rebecca Waters - Juan's Medium Gourmet Salsa - 2 pk
"I truly like the web site you have created and printed off several recipes. Also, because of your website I also ordered the enchilada sauce for us to try. Thanks again for the convenience you give your customers."
– Brenda McDonald - Juans Gourmet Enchilada Sauce - 2 pk
"I just thought it was one of the best fresh salsas that I've tasted in a long time. The product sells well."
– Tim Nash, Wine Experience - Juan's Supremo Habanero Hot Salsa - 2 pk
Juan O'Sullivan
Cumming, IA Copyright © 2019 Juan O'Sullivan. All Rights Reserved.
Web design & development by
Visionary Services, Inc.


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I am again here to share one Free PSD file, as I am keep sharing on Friday Freebie session. Today you can download a cool Looking Twitter Bird Icon PSD for free. The PSD was hosted on my other blog which is LazyBuddy.com.
The PSD is fully layered and can be easily customize. So don't wait, download it now and use it on your projects.
Download Twitter Bird Icon PSD Free
If you don't want to miss a single freebie, I recommend you to subscribe our Email Newsletter and get all the updates directly to your email. Below is the subscription box, just enter Your name Email ID and get updated. Thanks


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GUERRILLA INNOVATION: GAYmobile
It's been a while but this I couldn't resist. A new mobile operator in Denmark is targeting the Pink Krone. The GAYmobile.dk website offers the HOMOBILE service line which will be selling subtle ringtones that indicate a user's sexual preference (like the handkechief code?). Until the service comes online the company suggests that subscribers use songs like "I am what I am" or anything by ABBA as their ringtone. I could make a fortune selling my ringtones in Denmark. Oh yes, they say that they will donate 25% of their profits to the gaytrust.


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What is a Social Network?
We hear this term banded about the place but in case you are wondering what it means and you are too 'shy' to ask your children then here's some useful information from the NSPCC – a quick guide to social networking:
Social networking is the way people interact with each other online using websites, apps and games. It's a way of sharing things like photos. Children become more attracted to social networking when they move to Secondary school as they become more independent and want to become part of the social network of their new school, whether online or in person.
• Photos and images
• Message each other
• Message to a group of people (some may well be strangers)
• Chat with people using video
• Make voice calls
• Share content
• Meet new people and chat to strangers
• Play games
Most young people LOVE to social network because they can express themselves and socialise.
Keep an eye on your social networker.


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by Directory Info on 06-25-2017 05:00AM in Social Events
UAE Shipping Directory wishes you & your family Eid Mubarak
by Ali Khan on 08-04-2016 10:54AM in Social Events
It can be a difficult task for to find out a place for parking in Dubai especially in such situation when you are new in that area. You cannot feel satisfied when you pay an hourly fee, so for the facilitation now you can get the best services of parking in Dubai by paying fees on the hour and the fee has to be in dirham coins and can be paid by phone
by Directory Info on 07-07-2016 03:39PM in Social Events
Disclaimer: If you are aware of any inaccurate information, please report any listing from its page and provide us with the updated details.
You can also claim ownership of listing by clicking the 'Claim' option on each listing page – this will allow you to edit the current listing in our database.
UAE Ports & Shipping Directory © 2001-2019
All Rights Reserved.
All logos and trademarks in this site are property of their respective owners.


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Premium care through superior technology
Creating the premium practice means staying "ahead of the pack" in a number of areas, not the least of which being the technology aspect of ophthalmology.
A LASIK Seminar you can view at home in your spare time.
Anterior Segment Optical Coherence Tomography is one of the most powerful tools for the Corneal Surgeon.
TrueVision brought their 3D video system to the Wills Eye Operating Room. Dr. Lewis performed this DSEK surgery (corneal transplant for Fuchs/Endothelial Dystrophy).
LASIK no longer has an advantage over Staar's Visian ICL (Intraocular Contact Lens). Nearly half of the ICL's inserted in the US are now being done bilaterally.
James S. Lewis, MD, a pioneer in the use of Intense Pulsed Light (IPL) for treatment of dry eye in the philadelphia area is now the first practice to receive the Lipiflow System. LipiFlow is a FDA-Approved Dry Eye Treatment Read More
James S. Lewis, MD of James S. Lewis, MD PC in Elkins Park, PA Dr. Lewis patients are no longer required to purchase or use post-operative eye drops*. This eliminates costly co-pays and a visit to the pharmacy. In addition to cost, patients do not have to worry about timely and sterile instillation of these medications. Read More
The EPIC is a 5K camera, and can shoot a lovely 120 FPS at that, but to get 300 FPS
It's actually very easy to use. Touch screen menu make everything from frame rate to ISO to shutter speed and much much more all easily accessible via the very intuitive menu but more on that in my next RED post once that shoot it done.
Creating the premium practice means staying "ahead of the pack" in a number of areas, not the least of which being the technology aspect of ophthalmology.
Dr. James S. Lewis invites patients to the renovated Eye Institute of Salus University. This state-of-the-art North Philadelphia facility includes the James S. Lewis, MD Ophthalmology Suite specializing in cataract, cornea and LASIK Surgery.


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Ecwid and Square have teamed up to help you sell seamlessly online,
The integrated solution is available in the US, Canada, Australia and now in the UK!
Read on to learn more about the features of the integration, how it works and how Square can benefit your business.
Why Ecwid + Square?
Ecwid and Square give you the tools you need to run a successful business, both online and offline. You can sync inventory and product catalogs to save time, eliminate manual tasks and avoid overselling. Here are some benefits for using Square with Ecwid:
Accept payments anywhere: online and in person
Square Payments and POS options let you accept payments anywhere, whether you are thinking about opening a
Seamless automatic synchronization
With all of these new sales channels, you might be wondering how you're going to keep your inventory straight. Managing multiple storefronts is simple with
Ecwid and Square automatically update your inventory and order data across online and offline channels, thus streamlining store management and preventing issues like double entry and inventory discrepancies. You never have to track your physical and digital sales separately again.
Connect Ecwid and Square
Ready to get started? Here's how
There is no need to spend your valuable time on a lengthy setup process. You will be selling and accepting payments in just a few clicks:
To accept payments online
Accept Visa, MasterCard, Discover, and American Express for 1.75% per tap, chip, or swipe. Pay 2.5% per transaction for online sales in the UK.
To set up, login to your Ecwid Control Panel and connect with Square. Simply navigate to Settings → Payment→ Credit Card, select [Credit Card: Square] in the drop down menu and select Enable. Then click, Connect and Enable Square account and you will be ready to accept Square payments through your Ecwid store.
If you don't have an Ecwid account, click here to create an account and start selling.
Note: You must sign up for a Square account to use Square payments.
To accept payments offline with POS
Depending on where you are selling, you will want to make sure you have the right equipment. If you are selling on the go
Note: POS is available for Ecwid Unlimited Plan users.
Then login to your Ecwid Control Panel and navigate to Settings → POS, select Square, then click, Sync your inventory.
This will enable you to sell both online and offline with automatic inventory and product updates, so you can track everything in one place.
If you already are using Square, simply log into your Square account, find Apps and look for Ecwid's listing in the
You already have products, customers, and are generating revenue from your sales in the online world, so why not take your business one step further with an Ecwid and Square integrated solution? With Ecwid and Square you can sell your products from home, at a conference, or anywhere you choose!
Connect Ecwid and Square
If you have questions, please feel free to leave them in the comments section below or ask our Customer Care team.
Happy selling!


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Originally Posted by TReX
ASUS is among the leading players in the router market, constantly pushing the envelope in terms of performance and design. The latest high-end entry in its lineup is no different. The new RT-AX88U, which is also ASUS' first 802.11ax offering, promises transfer speeds of up 5,962Mbps over Wi-Fi and features sharp edges, massive antennas and gold accents -- just what you would expect out of an
Originally Posted by Omega
Íîâèíêè îò Asus:
ASUS Blue Cave
AC2600 Dual-Band Smart Wi-Fi Router
ASUS Lyra
AC2200 Tri-Band Whole-Home Wi-Fi System Mesh Network
Updated 03-06-2017 at 23:47 by Omega
Originally Posted by TReX
Óñòàíîâêà SSD â ðîóòåð )
~ Specs as follows ~
1). CPU ~ QCA IPQ8065 1.7GHz dual core Krait 300 CPU
2). WiFi ~ 2x QCA9984 4X4:4 Wave2 MU-MIMO 160MHz for both 2.4/5GHz (80MHz+80MHz)
(Specs is ready but 160 drivers are not ready yet)
3). LAN Switch ~ (LAN:1 x RLT8370MB + WAN: 2 x QCA8033)
4). Fully hardware NAT dual WAN for Internet
Originally Posted by Omega
Asus OnHub SRT-AC1900
Dual-band Wireless-AC1900 Smart Router
Intelligent internal antenna array pushes 802.11ac Wi-Fi speeds up to 1900 MbpsThe Google On app for Android and iOS allows simple setup and managementASUS-exclusive Wave Control lets you prioritize traffic to devices with a wave of your handSmart high-performance internal antennas,
Updated 10-11-2015 at 01:35 by Omega
Originally Posted by Omega
Asus unveils a monster of a Wi-Fi router, the RT-AC5300
If you think the Asus RT-AC3200 is overkill, you have no idea. The networking vendor announced today
at IFA its latest router, the RT-AC5300, which has much higher Wi-Fi specs than its older brother.
Similar to the RT-AC3200, the RT-AC5300
Updated 03-11-2015 at 02:59 by Omega


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Mary Ann Johnson
NAMI Southern Arizona Board President
After working as a teacher and a human resources manager, Mary Ann Johnson has been a personal Financial Advisor for the past 18 years, currently with Ameriprise. She is also the proud Nana of four grandchildren. Mary Ann has been a member of the Board of Directors since 2010 and is currently serving as Board President. "I came to NAMI Southern Arizona through the Family-to-Family program and later taught the course twice. The people I have met through NAMI who have shared their struggles and blessings have shown me a way to live a positive life despite the challenges of living with a mental illness." Mary Ann is particularly proud of the work of the Board of Directors over the past few years in designing and beginning the implementation of a 3-year Strategic Plan. "I have been privileged to work with an exceptional group on the Board who are willing to give time and expertise to ensure that NAMI Southern Arizona will continue its work for all those who will benefit from these services in the future."


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The Outlook wasn't brilliant for the Democratic gang that day:
The score stood 50 to 46, with but one month more to play.
And then when Maddow died on air, and Cutter did the same,
A sickly silence fell upon the donors of the gang.
A moderate few got up to go in long delayed realization. Those who only whined
Clung to that stupidity which springs eternal within the liberal mind;
They thought, if only Obama could get but a whack at Mitt-
We'd put up even money, now, with Obama in his chair.
But Alexrod preceded Obama, as did also Lawrence O'Donnell
And the former was a cuckoo and the latter was a ditz;
So upon that entitlement gathering multitude grimly watched the end of Obamacare,
For there seemed but little chance of Obama filling that empty chair.
But Alexrod let drive a bullshit lie that was believed only in states of blue,
And O'Donnell, the much enthralled, tore the cover off his claims of true;
And when the dust had lifted, and the men saw what had occurred,
There was Alexrod safe from facts and O'Donnell quite absurd.
Then from 500 people still watching MSNBC there rose a vapid yell;
It rumbled through the entitlement offices, it rattled like an Embassy being shelled;
It knocked upon the mountain that is Obamacare and recoiled those getting welfare
For Obama, mighty Obama, was advancing to his chair
There was ease in Obama's manner as he strutted into his place;
There was arrogance in Obama's bearing and a smirk on Obama's face.
And when, responding to the cheers, he smugly glared at Mitt,
No stranger in the crowd could doubt 'twas Obama and his bullshit.
Ten million eyes were on him as he showered himself with praise;
Five thousand tongues applauded while the Middle East was ablaze.
Then while the withering moderator let the first question zip,
Defiance gleamed in Obama's eye, a sneer curled Obama's lip.
And now fact laden challenger came knowledgably across the stage,
And Obama stood a-watching it in haughty grandeur with nothing to read off a page
Close by the pompous Pres the questions unheeded sped-
"That ain't my style," said Obama. "Strike one," the public said.
From the newsrooms, somber with pundits, there went up muffled squalls,
As if the tingle up Matthews' leg had turned to a kick in the balls
"Kill him! Kill the Republican!" shouted someone out in Twitter land;
And in deluded fog Obama's Secret Service did not raise a hand.
(Probably busy looking for coke and hookers in South America).
With a smug look of tyrannical charm great Obama's message resembled Lenin,
And Joe replace the rising tumult with horror of his laughter and grin,
Barry signaled to the audience, and once more his lies they flew
But public still ignored them, and the polls said, "Strike two."
"Fraud!" cried Candy, her facts all wrong, and echo of MSM answered fraud;
But one scornful look from Obama and the blind followers were awed.
They saw his face grow stern and cold, they saw his distorted facts strain,
And they knew that Obama wouldn't let that debate go by again.
The sneer is great from Obama's lip, his teeth are clenched in hate;
He pounds with cruel violence his fist and his lies he does restate.
And now Romney holds the truth and the public, and now out to vote they go,
And now the myth of Obama is shattered by an economy that is slow.
Oh everywhere in this favored nation the sun is shining bright;
The jobs are returning everywhere, and everywhere hearts are light,
And everywhere people are prosperous, and everywhere Democrats pout;
For there is no joy in the welfare office – mighty Obama has struck out.


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indicates every 2 miles
Charlton Station-paddington Station Cycle Route
|Cycle Route Details
|Route Name
|Charlton Station-paddington Station
|Distance : miles (km)
|11.77 (18.94)
|Posted By
|conall askew
|Calories Burned
|(login and provide your weight for more accurate calories burned)
|Cycle Steps
|A good route for swift commute from South-East London to the City.
|Landmarks/Pubs for lunch/Sites to see?
|house of commons, london eye, buckingham palace
|Near By Cycle Shops
|Charlton Halfords
|Action Bikes
|Dial A Bike
|Fluid Cycles
|Cycles UK
|The Bike Rooms
|Old Kent Road Halfords
|Cycling Clubs located in this region
|Anerley Bicycle Club
|Herne Hill Youth Cycling Club
|Kenton Road Club
|Kingston Phoenix Road Club
|London Clarion Cycle Club
|The Trax
|Woolwich CC
|Your comments?
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