Direct measurement of DNA-mediated adhesion between lipid bilayers

Multivalent interactions between deformable mesoscopic units are ubiquitous in biology, where membrane macromolecules mediate the interactions between neighbouring living cells and between cells and solid substrates. Lately, analogous artificial materials have been synthesised by functionalising the outer surface of compliant Brownian units, for example emulsion droplets and lipid vesicles, with selective linkers, in particular short DNA sequences. This development extended the range of applicability of DNA as a selective glue, originally applied to solid nano and colloidal particles. On very deformable lipid vesicles, the coupling between statistical effects of multivalent interactions and mechanical deformation of the membranes gives rise to complex emergent behaviours, as we recently contributed to demonstrate [Parolini et al., Nature Communications, 2015, 6, 5948]. Several aspects of the complex phenomenology observed in these systems still lack a quantitative experimental characterisation and fundamental understanding. Here we focus on the DNA-mediated multivalent interactions of a single liposome adhering to a flat supported bilayer. This simplified geometry enables the estimate of the membrane tension induced by the DNA-mediated adhesive forces acting on the liposome. Our experimental investigation is completed by morphological measurements and the characterisation of the DNA-melting transition, probed by in-situ F\"{o}rster Resonant Energy Transfer spectroscopy. Experimental results are compared with the predictions of an analytical theory that couples the deformation of the vesicle to a full description of the statistical mechanics of mobile linkers. With at most one fitting parameter, our theory is capable of semi-quantitatively matching experimental data, confirming the quality of the underlying assumptions.Comment: 16 pages, 7 figure

Assessment of landfill leachate biodegradability and treatability by means of allochthonous and autochthonous biomasses

The biodegradability and treatability of a young (3 years old) municipal landfill leachate was evaluated by means of chemical oxygen demand (COD) fractionation tests, based on respirometric techniques. The tests were performed using two different biomasses: one cultivated from the raw leachate (autochthonous biomass) and the other collected from a conventional municipal wastewater treatment plant after its acclimation to leachate (allochthonous biomass). The long term performances of the two biomasses were also studied. The results demonstrated that the amount of biodegradable COD in the leachate was strictly dependent on the biomass that was used to perform the fractionation tests. Using the autochthonous biomass, the amount of biodegradable organic substrate resulted in approximately 75% of the total COD, whereas it was close to 40% in the case of the allochthonous biomass, indicating the capacity of the autochthonous biomass to degrade a higher amount of organic compounds present in the leachate. The autochthonous biomass was characterized by higher biological activity and heterotrophic active fraction (14% vs 7%), whereas the activity of the allochthonous biomass was significantly affected by inhibitory compounds in the leachate, resulting in a lower respiration rate (SOUR = 13 mg O2 gVSS-1 h-1 vs 37 mg O2 gVSS-1 h-1). The long-term performance of the autochthonous and allochthonous biomasses indicated that the former was more suitable for the treatment of raw landfill leachate, ensuring higher removal performance towards the organic pollutants

Dermoscopy and methyl aminolevulinate: A study for detection and evaluation of field cancerization

Actinic keratosis (AK) is a keratinocyte intraepidermal neoplasia UV light – induced that frequently appears in sun-exposed areas of the skin. Although historically AK was de fi ned as “ precancerous ” , actually it is considered as the earliest stage of squamous cell carcinoma (SCC) in situ. Since AKs can progress into invasive SCC, their treatment isrecommended. AKsrarely developasa singlelesion;usually multiplelesions commonly affect anen- tire area of chronically actinic damaged skin. This has led to the concept of “ fi eld cancerization ” , an area chroni- cally sun-exposed that surrounds peripherally visible lesions, in which are individualized subclinical alterations. One of the main principles endpoint in the management of AKs is the evaluation and the treatment of fi eld cancerization. In this view, in order to detect and quantify fi eld cancerization, we employed a method based on the topical application of methyl aminolevulinate (MAL) and the detection of the fl uorescence emitted by its metabolite Protoporphyrin IX (PpIX); then, considering the extension and the intensity of measured fl uores- cence, we create a score of fi eld cancerization. The results show that patients underwent to daylight PDT had a reduction of total score, from T0 to T2. Whereas in the group untreated we observed a stability of total score or a slightly worse. So, the method and the score used allows to evaluate with a good approximation the dimension of fi eld cancerization and show the modi fi cation of it after treatment

A quantum mechanical insight into SN2 reactions: Semiclassical initial value representation calculations of vibrational features of the Cl−^---CH3_3Cl pre-reaction complex with the VENUS suite of codes

The role of vibrational excitation of reactants in driving reactions involving polyatomic species has been often studied by means of classical or quasi-classical trajectory simulations. We propose a different approach based on investigation of vibrational features of the Cl$^-$--CH$_3$Cl pre-reaction complex for the Cl$^-$ + CH$_3$Cl SN$_2$ reaction. We present vibrational power spectra and frequency estimates for the title pre-reaction complex calculated at the level of classical, semiclassical, and second-order vibrational perturbation theory on a pre-existing analytical potential energy surface. The main goals of the paper are the study of anharmonic effects and understanding of vibrational couplings that permit energy transfer between the collisional kinetic energy and the internal vibrations of the reactants. We provide both classical and quantum pictures of intermode couplings and show that the SN2 mechanism is favored by the coupling of a C--Cl bend involving the Cl$^-$ projectile with the CH$_3$ rocking motion of the target molecule. We also illustrate how the routines needed for semiclassical vibrational spectroscopy simulations can be interfaced in a user-friendly way to pre-existing molecular dynamics software. In particular, we present an implementation of semiclassical spectroscopy into the VENUS suite of codes, thus providing a useful computational tool for users who are not experts of semiclassical dynamics

Seasonal Training-Load Quantification in Elite English Premier League Soccer Players

Purpose: To quantify the seasonal training load completed by professional soccer players of the English Premier League. Methods: Thirty players were sampled (using GPS, heart rate, and rating of perceived exertion [RPE]) during the daily training sessions of the 2011–12 preseason and in-season period. Preseason data were analyzed across 6 × 1-wk microcycles. In-season data were analyzed across 6 × 6-wk mesocycle blocks and 3 × 1-wk microcycles at start, midpoint, and end-time points. Data were also analyzed with respect to number of days before a match. Results: Typical daily training load (ie, total distance, high-speed distance, percent maximal heart rate [%HRmax], RPE load) did not differ during each week of the preseason phase. However, daily total distance covered was 1304 (95% CI 434–2174) m greater in the 1st mesocycle than in the 6th. %HRmax values were also greater (3.3%, 1.3–5.4%) in the 3rd mesocycle than in the first. Furthermore, training load was lower on the day before match (MD-1) than 2 (MD-2) to 5 (MD-5) d before a match, although no difference was apparent between these latter time points. Conclusions: The authors provide the 1st report of seasonal training load in elite soccer players and observed that periodization of training load was typically confined to MD-1 (regardless of mesocycle), whereas no differences were apparent during MD-2 to MD-5. Future studies should evaluate whether this loading and periodization are facilitative of optimal training adaptations and match-day performance

Computational modelling of emboli travel trajectories in cerebral arteries: Influence of microembolic particle size and density

This article has been made available through the Brunel Open Access Publishing Fund.Ischaemic stroke is responsible for up to 80 % of stroke cases. Prevention of the reoccurrence of ischaemic attack or stroke for patients who survived the first symptoms is the major treatment target. Accurate diagnosis of the emboli source for a specific infarction lesion is very important for a better treatment for the patient. However, due to the complex blood flow patterns in the cerebral arterial network, little is known so far of the embolic particle flow trajectory and its behaviour in such a complex flow field. The present study aims to study the trajectories of embolic particles released from carotid arteries and basilar artery in a cerebral arterial network and the influence of particle size, mass and release location to the particle distributions, by computational modelling. The cerebral arterial network model, which includes major arteries in the circle of Willis and several generations of branches from them, was generated from MRI images. Particles with diameters of 200, 500 and 800 μ m and densities of 800, 1,030 and 1,300 kg/m 3 were released in the vessel's central and near-wall regions. A fully coupled scheme of particle and blood flow in a computational fluid dynamics software ANASYS CFX 13 was used in the simulations. The results show that heavy particles (density large than blood or a diameter larger than 500 μ m) normally have small travel speeds in arteries; larger or lighter embolic particles are more likely to travel to large branches in cerebral arteries. In certain cases, all large particles go to the middle cerebral arteries; large particles with higher travel speeds in large arteries are likely to travel at more complex and tortuous trajectories; emboli raised from the basilar artery will only exit the model from branches of basilar artery and posterior cerebral arteries. A modified Circle of Willis configuration can have significant influence on particle distributions. The local branch patterns of internal carotid artery to middle cerebral artery and anterior communicating artery can have large impact on such distributions. © 2014 The Author(s)

Beam manipulation for resonant plasma wakefield acceleration

Plasma-based acceleration has already proved the ability to reach ultra-high accelerating gradients. However the step towards the realization of a plasma-based accelerator still requires some e ff ort to guarantee high brightness beams, stability and reliability. A significant improvement in the efficiency of PWFA has been demonstrated so far accelerating a witness bunch in the wake of a higher charge driver bunch. The transformer ratio, therefore the energy transfer from the driver to the witness beam, can be increased by resonantly exciting the plasma with a properly pre-shaped drive electron beam. Theoretical and experimental studies of beam manipulation for resonant PWFA will be presented her

Fragmentation and depolymerization of non-covalently bonded filaments.

Protein molecules often self-assemble by means of non-covalent physical bonds to form extended filaments, such as amyloids, F-actin, intermediate filaments, and many others. The kinetics of filament growth is limited by the disassembly rate, at which inter-protein bonds break due to the thermal motion. Existing models often assume that the thermal dissociation of subunits occurs uniformly along the filament, or even preferentially in the middle, while the well-known propensity of F-actin to depolymerize from one end is mediated by biochemical factors. Here, we show for a very general (and generic) model, using Brownian dynamics simulations and theory, that the breakup location along the filament is strongly controlled by the asymmetry of the binding force about the minimum, as well as by the bending stiffness of the filament. We provide the basic connection between the features of the interaction potential between subunits and the breakup topology. With central-force (that is, fully flexible) bonds, the breakup rate is always maximum in the middle of the chain, whereas for semiflexible or stiff filaments this rate is either a minimum in the middle or flat. The emerging framework provides a unifying understanding of biopolymer fragmentation and depolymerization and recovers earlier results in its different limits.This work has been supported by the Ernest Oppenheimer Fellowship at Cambridge (AZ, LD) and by the Technische Universität München Institute for Advanced Study, funded by the German Excellence Initiative and the EU 7th Framework Programme under grant agreement nr. 291763 (AZ). LD also acknowledges the Marie Curie ITNCOMPLOIDS grant no. 234810.This is the accepted manuscript. The final version is available at http://scitation.aip.org/content/aip/journal/jcp/142/11/10.1063/1.4914925