Identification of in vitro phosphorylation sites in the growth cone protein SCG10. Effect Of phosphorylation site mutants on microtubule-destabilizing activity

SCG10 is a neuron-specific, membrane-associated protein that is highly concentrated in growth cones of developing neurons. Previous studies have suggested that it is a regulator of microtubule dynamics and that it may influence microtubule polymerization in growth cones. Here, we demonstrate that in vivo, SCG10 exists in both phosphorylated and unphosphorylated forms. By two-dimensional gel electrophoresis, two phosphoisoforms were detected in neonatal rat brain. Using in vitro phosphorylated recombinant protein, four phosphorylation sites were identified in the SCG10 sequence. Ser-50 and Ser-97 were the target sites for protein kinase A, Ser-62 and Ser-73 for mitogen-activated protein kinase and Ser-73 for cyclin-dependent kinase. We also show that overexpression of SCG10 induces a disruption of the microtubule network in COS-7 cells. By expressing different phosphorylation site mutants, we have dissected the roles of the individual phosphorylation sites in regulating its microtubule-destabilizing activity. We show that nonphosphorylatable mutants have increased activity, whereas mutants in which phosphorylation is mimicked by serine-to-aspartate substitutions have decreased activity. These data suggest that the microtubule-destabilizing activity of SCG10 is regulated by phosphorylation, and that SCG10 may link signal transduction of growth or guidance cues involving serine/threonine protein kinases to alterations of microtubule dynamics in the growth cone.     

Biodegradable hollow microfibres to produce bioactive scaffolds

Biodegradable hollow microfibres containing particles loaded with specific active agents can be potentially employed to produce a special kind of substrate for tissue engineering, able to function as a scaffold and at the same time to act as a drug‐releasing system. Biodegradable hollow microfibres based on poly(lactic acid) were produced by a dry–wet spinning procedure. Drug‐loaded microparticles were prepared by a simple oil‐in‐water emulsion and entrapped inside the fibres. The morphology of both fibres and particles was investigated by scanning electron microscopy. The mechanical and thermal properties of the fibres were investigated by tensile tests and differential scanning calorimetry. In vitro tests were performed to evaluate the release of the drug from the fibres loaded with the particles Copyright © 2004 Society of Chemical Industry     

Inorganic–organic hybrid materials with zirconium oxoclusters as protective coatings on aluminium alloys

Inorganic–organic hybrid materials are attracting a strong scientific interest mainly for their outstanding inherent mechanical and thermal properties, which can be traced back to the intimate coupling of both inorganic and organic components. By carefully choosing the experimental parameters used for their synthesis, chemically and thermally stable acrylate-based hybrid material embedding the zirconium oxocluster Zr₄O₄(OMc)₁₂, where OMcCH₂C(CH₃)C(O)O, can be deposited as UV-cured films on aluminium alloys.

In particular, the molar ratios between the oxocluster and the monomer, the polymerisation time, the amount of photo-initiator and the deposition conditions, by using an home-made spray-coating equipment, were optimised in order to obtain the best performing layers in terms of transparency and hardness to coat aluminium alloy (AA1050, AA6060 and AA2024) sheets. Furthermore, it was also evaluated whether the hybrid coatings behave as barrier to corrosion.

Several coated samples were prepared and characterised. Environmental scanning electronic microscopy (ESEM) and scratch test were used to investigate the morphology of the films and to evaluate their scratch resistance, respectively. Electrochemical impedance spectroscopy (EIS) was performed in order to evaluate if the coatings actually protect the metallic substrate from corrosion.

In order to measure shear storage modulus (G′) and loss modulus (G″) of the materials used for coatings, bulk samples were also obtained by UV-curing of the precursors solution. Dynamical mechanical thermal analysis (DMTA) was performed in shear mode on cured disks of both the hybrid materials and pristine polymer for comparison. The values of T_g were read off as the temperatures of peak of loss modulus. The length and mass of all the samples were measured before and after the DMTA analysis, so that the shrinkage of the materials in that temperature range was exactly evaluated.     

Procedural memory stimulation in Alzheimer''s disease: impact of a training programme

The operative results of 23 patients with a specific or unspecific spondylodiscitis were documented over 2 years after the focus of the inflammation had been eradicated, bone chip had been interposed and a CDH instrumentation had been performed by an anterior approach only. These outcomes were compared with the results of 32 patients in whom the focus had been removed and the defect had been filled with bone graft from an anterior approach, followed by stabilisation with CD instrumentation through an additional dorsal approach. In the cases where CDH instrumentation was applied, the range of fusion averaged 1.3 segments. This was clearly less extensive than in dorsoventral stabilisation, in which on average 3.5 segments were fused. In 47 of 55 cases mobilisation was achieved without orthesis. Eight months after the operations bony fusion could be observed radiologically in all patients. The mean preoperative kyphotic angle of the affected segments was 14.4 degrees, compared to 4 degrees after the operation. The mean loss of reposition was measured to be about 2.7 degrees in both groups. Average operation time and blood loss were about 50% higher in the patients treated dorsoventrally. We conclude that even in the case of florid spondylodiscitis, a short-range anterior fusion of the affected spinal segment may be performed by use of a stable-angle implant without an increased risk of infection-related loosening.     

Power Dissipation in Spintronic Devices Out of Thermodynamic Equilibrium

Quantum limits of power dissipation in spintronic computing are estimated. A computing element composed of a single electron in a quantum dot is considered. Dynamics of its spin due to external magnetic field and interaction with adjacent dots are described via the Bloch equations. Spin relaxation due to magnetic noise from various sources is described as coupling to a reservoir. Resulting dissipation of energy is calculated and is shown to be much less than the thermal limit, ∼kT per bit, if the rate of spin relaxation is much slower than the switching rate. Clues on how to engineer an energy efficient spintronic device are provided.     

Hydrocarbon exploration risk evaluation through uncertainty and sensitivity analyses techniques

The evaluation of the exploration risk in the oil industry is a fundamental component of the decision process related to the exploratory phase. In this paper the two basic components of the exploratory risk: trap geometry and trapped hydrocarbon quantities (fluid), are compounded in a single coherent uncertainty and sensitivity approach. The results clarify that the model geometry influences each Petroleum System Modeling step and that the geometric uncertainty is correlated with the fluid uncertainty. The geometric uncertainty evaluation makes use of geostatistical techniques that produce a number of possible realizations of the trap geometry, all compatible with available data. The evaluation of the fluid uncertainty, through a Monte Carlo methodology, allows us to compute the possible quantities of oil and gas, generated in a basin and migrated from the hydrocarbon source location to each single trap. The final result is the probability distribution of oil and gas for each trap in the basin, together with other useful indicators like: the hydrocarbon filling probability map, the closure probability map, the drainage area probability map, the spilling paths probabilities, the trap-filling scenarios.     

Cerebral dominance and asynchrony between bimanual two-dimensional movements.

The asynchrony of bimanual movements was investigated. Right- and left-handers traced simple geometrical patterns (ellipses) continuously with both hands. All combinations of the direction of rotation in each hand were executed at different rhythms. Geometrically, performances were largely independent of manual dominance. However, by comparing the passage times at homologous positions, the authors found that the dominant hand led the nondominant one by about 25 msec. The asynchrony was affected by neither movement type nor rhythm. The variability of the asynchrony varied along the trajectory, with well-defined maxima and minima. The variability profiles for movements that engaged homologous muscles differed markedly from those that engaged nonhomologous muscles. The authors discuss the hypotheses that bimanual periodic movements are timed by a lateralized functional module and that asynchrony is due to the necessity of transmitting time-keeping information to the other hemisphere. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Toward automatic phenotyping of developing embryos from videos.

We describe a trainable system for analyzing videos of developing C. elegans embryos. The system automatically detects, segments, and locates cells and nuclei in microscopic images. The system was designed as the central component of a fully automated phenotyping system. The system contains three modules 1) a convolutional network trained to classify each pixel into five categories: cell wall, cytoplasm, nucleus membrane, nucleus, outside medium; 2) an energy-based model, which cleans up the output of the convolutional network by learning local consistency constraints that must be satisfied by label images; 3) a set of elastic models of the embryo at various stages of development that are matched to the label images.     

Algorithms for multiprocessor scheduling with two job lengths and allocation restrictions

A variant of the High Multiplicity Multiprocessor Scheduling Problem with C job lengths is considered, in which jobs can be processed only by machines not greater than a given index. When C=2, polynomial algorithms are proposed, for the feasibility version of the problem and for maximizing the number of scheduled jobs.