NGF Enhances CGRP Release Evoked by Capsaicin from Rat Trigeminal Neurons: Differential Inhibition by SNAP-25-Cleaving Proteases

Nerve growth factor (NGF) is known to intensify pain in various ways, so perturbing pertinent effects without negating its essential influences on neuronal functions could help the search for much-needed analgesics. Towards this goal, cultured neurons from neonatal rat trigeminal ganglia—a locus for craniofacial sensory nerves—were used to examine how NGF affects the Ca²⁺-dependent release of a pain mediator, calcitonin gene-related peptide (CGRP), that is triggered by activating a key signal transducer, transient receptor potential vanilloid 1 (TRPV1) with capsaicin (CAP). Measurements utilised neurons fed with or deprived of NGF for 2 days. Acute re-introduction of NGF induced Ca²⁺-dependent CGRP exocytosis that was inhibited by botulinum neurotoxin type A (BoNT/A) or a chimera of/E and/A (/EA), which truncated SNAP-25 (synaptosomal-associated protein with Mr = 25 k) at distinct sites. NGF additionally caused a Ca²⁺-independent enhancement of the neuropeptide release evoked by low concentrations (<100 nM) of CAP, but only marginally increased the peak response to ≥100 nM. Notably, BoNT/A inhibited CGRP exocytosis evoked by low but not high CAP concentrations, whereas/EA effectively reduced responses up to 1 µM CAP and inhibited to a greater extent its enhancement by NGF. In addition to establishing that sensitisation of sensory neurons to CAP by NGF is dependent on SNARE-mediated membrane fusion, insights were gleaned into the differential ability of two regions in the C-terminus of SNAP-25 (181–197 and 198–206) to support CAP-evoked Ca²⁺-dependent exocytosis at different intensities of stimulation.     

Reversible Amorphous‐to‐Amorphous Transitions in Chalcogenide Films: Correlating Changes in Structure and Optical Properties

Structural transitions in materials are accompanied by appreciable and exploitable changes in physical‐chemical properties. Whereas reversible optically‐driven atomistic changes in crystal‐to‐amorphous transitions are generally known and exploited in applications, the nature of the corresponding polyamorphic transitions between two structurally distinct meta‐stable amorphous phases is an unexplored theme. Direct experimental evidence is reported for the nature of the atomistic changes during fully reversible amorphous‐to‐amorphous switching between two individual states in the non‐crystalline As₅₀Se₅₀ films prepared by pulsed‐laser deposition and consequent changes in optical properties. Combination of surface sensitive X‐ray photoelectron spectroscopy and spectroscopic ellipsometry show that the near‐bandgap energy illumination and annealing induce reversible switching in the material's structure by local bonding rearrangements. This is accompanied by switching in refractive index between two well‐defined states. Exploiting the pluralism of distinct structural states in a disordered solid can provide new insights into the data storage in emerging optical memory and photonic applications.     

Oblique impact simulations of high strength agglomerates

Different types of particle compounds like concrete particles can be considered as a model material of high strength agglomerates. It is necessary to investigate and understand fracture behaviour of these agglomerates in order to avoid breakage during storage, handling and transportation. The aim of the research is to examine the comminution behaviour of high strength agglomerates during oblique impact loadings.A two dimensional finite element analysis has been carried out to understand stress pattern distributions before crack initiation. Then a two dimensional discrete element method has been applied to study the fragmentation behaviour of the agglomerates. Concrete particles of B35 strength category have been chosen to represent the high strength agglomerates.Analysis is done with oblique impact loadings at different velocities from 7.7 to 180 m/s. The stressing conditions comprise low flow rate transportation and handling to high speed impacts during fall down in bunker, stock piles, ship loading or stressing in crushers and mill operations. Particle size distributions and new surface generation have also been evaluated in the paper.It is shown that at higher velocities, particle size distributions are identical to each other regardless of impact angle. Increasing impact velocity does not necessarily produce more new surfaces after certain velocity limit.     

A Feature Selection Algorithm for the Regularization of Neuron Models

This paper presents a novel training method for estimating the parameters of retina models, such as integrate-and-fire (IF) or Poisson based. The presented models are constructed using a set of linear and nonlinear filters, which are described by basis functions and Taylor polynomials, respectively. This approach allows for the identification of a set of features that can be used for reproducing retina responses. By using the Bayesian-Laplace feature selection algorithm herein proposed, an efficient model with a reduced set of parameters is achieved. Experimental results show that the proposed algorithm is able to remove unimportant features while still accurately reproducing retina responses. These results also show that the IF model is able to mimic the retina visual processing system using less parameters than the Poisson-based model.     

Energy absorption during compression and impact of dry elastic-plastic spherical granules

The discrete modelling and understanding of the particle dynamics in fluidized bed apparatuses, mixers, mills and others are based on the knowledge about the physical properties of particles and their mechanical behaviour during slow, fast and repeated stressing. In this paper model parameters (modulus of elasticity, stiffness, yield pressure, restitution coefficient and strength) of spherical granules (γ-Al₂O₃, zeolites 4A and 13X, sodium benzoate) with different mechanical behaviour have been measured by single particle compression and impact tests. Starting with the elastic compression behaviour of granules as described by Hertz theory, a new contact model was developed to describe the force-displacement behaviour of elastic-plastic granules. The aim of this work is to understand the energy absorption during compression (slow stressing velocity of 0.02 mm/s) and impact (the impact velocity of 0.5–4.5 m/s) of granules. For all examined granules the estimated energy absorption during the impact is found to be far lower than that during compression. Moreover, the measured restitution coefficient is independent of the impact velocity in the examined range and independent of the load intensity by compression (i.e. maximum compressive load). In the case of repeated loading with a constant load amplitude, the granules show cyclic hardening with increasing restitution coefficient up to a certain saturation in the plastic deformation. A model was proposed to describe the increase of the contact stiffness with the number of cycles. When the load amplitude is subsequently increased, further plastic deformation takes place and the restitution coefficient strongly decreases.     

Development of a non-destructive micro-analytical method for stable carbon isotope analysis of transmission electron microscope (TEM) samples

The biogenicity of ancient morphological microfossil-like objects can be established by linking morphological (e.g. cell remnants and extracellular polymeric matrix) and chemical (e.g. isotopes, biomarkers and biominerals) evidence indicative of microorganisms or microbial activity. We have developed a non-destructive micro-analytical ion beam system capable of measuring with high spatial resolution the stable carbon isotope ratios of thin samples used for transmission electron microscopy. The technique is based on elastic scattering of alpha particles with an energy of 2.751 MeV. At this energy the ¹³C cross section is enhanced relative to the pure Rutherford cross section for ¹³C, whereas the ¹²C cross section is reduced relative to its pure Rutherford cross section. Here we report the initial results of this experimental approach used to characterize ultramicrotomed sections of sulfur-embedded graphite and microbial cells.     

Projective moment invariants

The paper is devoted to the moment invariants with respect to projective transform. It has been a common belief that such invariants do not exist. We show that projective moment invariants exist in a form of infinite series containing moments with positive as well as negative indices.     

Detection of phosphorylated peptides in proteomic analyses using microfluidic compact disk technology

A compact disk (CD)-based microfluidic method for selective detection of phosphopeptides by mass spectrometry is described. It combines immobilized metal affinity chromatography (IMAC) and enzymatic dephosphorylation. Phosphoproteins are digested with trypsin and processed on the CD using nanoliter scale IMAC with and without subsequent in situ alkaline phosphatase treatment. This is followed by on-CD matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Dephosphorylation of the IMAC-enriched peptides allows selective phosphopeptide detection based on the differential mass maps generated (mass shifts of 80 Da or multiples of 80 Da). The CD contains 96 microstructures, each with a 16 nL IMAC microfluidic column. Movement of liquid is controlled by differential spinning of the disk. Up to 48 samples are distributed onto the CD in two equal sets. One set is for phosphopeptide enrichment only, the other for identical phosphopeptide enrichment but combined with in situ dephosphorylation. Peptides are eluted from the columns directly into MALDI target areas, still on the CD, using a solvent containing the MALDI matrix. After crystallization, the CD is inserted into a MALDI mass spectrometer for analysis down to the femtomole level. The average success rate in phosphopeptide detection is over 90%. Applied to noncharacterized samples, the method identified two novel phosphorylation sites, Thr 735 and Ser 737, in the ligand-binding domain of the human mineralocorticoid receptor.