The role of histone arginine methylation in gene expression of airway smooth muscle cells in asthma

Introduction and objectives: Asthma is estimated to affect at least 300 million people globally. About 25% of the patients do not respond to therapy; therefore we need to develop novel treatments. ASM cells have a crucial role in asthma, contributing to airway remodelling, inflammation and airflow obstruction. We have previously shown that epigenetic histone modifications, particularly histone lysine acetylation and methylation regulate the secretion of inflammatory mediators from ASM cells. Here we tested the hypothesis that histone arginine changes are also involved. Protein arginine N-methyltransferases (PRMTs) are the enzymes which catalyse histone arginine methylation (HRme, the addition of a methyl group to arginine residues on the N-terminal tails of histones), and inhibiting them represents a strategy to reduce the secretion of inflammatory mediators from ASM cells. Methods: Studies were performed in cultured human ASM cells from asthmatic and non-asthmatic donors at passage 6. PRMT expression in human ASM cells was investigated by qPCR. Protein levels of four PRMTs in human ASM cells were investigated by western blotting. The effect of inhibiting PRMTs on the secretion of eotaxin, IL-6, CXCL8 and IP-10 from healthy ASM cells, under basal conditions and following stimulation with TNF-α (1ng/ml), was investigated by ELISA. Results: We found that ASM cells express the PRMT1, PRMT2, PRMT3, CARM1, PRMT5, PRMT6, PRMT7 and FBX011 mRNA and PRMT1, CARM1, PRMT5, and PRMT6 protein. The analysis showed no difference in the levels of expression between cells isolated from asthmatic and non-asthmatic donors. Two PRMT inhibitors, namely TCE5003 – a PRMT1 inhibitor, and 217531 - a CARM1 inhibitor, significantly reduced the secretion of inflammatory mediators from ASM cells. Conclusions: ASM cells express a number of PRMTs at mRNA and protein levels. The inhibition of PRMTs results in the reduced secretion of inflammatory mediators from ASM cells. PRMTs may have an important role in regulating chemokine production from ASM cells in asthma, and are a promising target for future investigations in asthma

Use of VLF transmissions in the location and mapping of lightning-induced ionisation enhancements (LIEs)

Lightning-induced ionisation enhancements (LIEs) are usually produced by short (~ 1 s) bursts of energetic electrons precipitated from the radiation belts in the process of amplifying whistlers. During their short life (~ 30 s) LIEs diffract or otherwise modify stable transmissions of VLF waves propagating in the two-dimensional Earth-ionosphere waveguide. This causes perturbations (‘Trimpis’) on the same time scale in the phase and amplitude of these VLF waves. Unless the LIEs are large (> 100 km) and smoothly varying (e.g., Gaussian distribution of ionisation enhancement) in the horizontal directions, the LIEs need not be on the great circle path (GCP) from VLF transmitter to receiver to produce Trimpis. Large and smooth LIEs produce ‘GCP Trimpis’, while small or structured LIEs produce ‘echo Trimpis’. The two can usually be distinguished, if Trimpi phase and amplitude are monitored and if the Trimpis are observed at several frequencies or on two or more spaced receivers simultaneously. If only GCP Trimpis are considered, the causative LIEs can be located and mapped by geometric optics using a network of receivers of sufficient density (spacing ~ 100 km) and a few transmitters. Provided all Trimpis are identified as GCP, their mere detection is sufficient for location. This is equivalent to locating the LIE ‘shadows’ cast onto arrays of spaced receivers by two or more transmitters. If this GCP identification is not made, or is just assumed, location and mapping (size estimation) errors can be quite large. At VLF (λ ~ 15 km) this geometric optics approach cannot be used to study the horizontal fine structure of LIEs since LIEs producing GCP Trimpis have no fine structure. Small or structured LIEs cast a diffraction pattern onto an array of spaced receivers. If both the phase and amplitude perturbation of echo Trimpis are measured at each receiver of the array, holographic techniques can be used to reconstruct the two-dimensional map or image of the causative LIEs. It is shown that, for a single system of one transmitter and a receiver array, this allows high resolution (~ 10 km) in the azimuthal dimension only. Equally high resolution in both horizontal dimensions can be achieved with two orthogonal systems. This technique works equally well on GCP Trimpis to map the causative LIEs (which are large and structureless) without incurring location errors thereby

The role of histone arginine methylation in gene expression of airway smooth muscle cells in asthma

Introduction and objectives: Asthma is estimated to affect at least 300 million people globally. About 25% of the patients do not respond to therapy; therefore we need to develop novel treatments. ASM cells have a crucial role in asthma, contributing to airway remodelling, inflammation and airflow obstruction. We have previously shown that epigenetic histone modifications, particularly histone lysine acetylation and methylation regulate the secretion of inflammatory mediators from ASM cells. Here we tested the hypothesis that histone arginine changes are also involved. Protein arginine N-methyltransferases (PRMTs) are the enzymes which catalyse histone arginine methylation (HRme, the addition of a methyl group to arginine residues on the N-terminal tails of histones), and inhibiting them represents a strategy to reduce the secretion of inflammatory mediators from ASM cells. Methods: Studies were performed in cultured human ASM cells from asthmatic and non-asthmatic donors at passage 6. PRMT expression in human ASM cells was investigated by qPCR. Protein levels of four PRMTs in human ASM cells were investigated by western blotting. The effect of inhibiting PRMTs on the secretion of eotaxin, IL-6, CXCL8 and IP-10 from healthy ASM cells, under basal conditions and following stimulation with TNF-α (1ng/ml), was investigated by ELISA. Results: We found that ASM cells express the PRMT1, PRMT2, PRMT3, CARM1, PRMT5, PRMT6, PRMT7 and FBX011 mRNA and PRMT1, CARM1, PRMT5, and PRMT6 protein. The analysis showed no difference in the levels of expression between cells isolated from asthmatic and non-asthmatic donors. Two PRMT inhibitors, namely TCE5003 – a PRMT1 inhibitor, and 217531 - a CARM1 inhibitor, significantly reduced the secretion of inflammatory mediators from ASM cells. Conclusions: ASM cells express a number of PRMTs at mRNA and protein levels. The inhibition of PRMTs results in the reduced secretion of inflammatory mediators from ASM cells. PRMTs may have an important role in regulating chemokine production from ASM cells in asthma, and are a promising target for future investigations in asthma

World-wide lightning location using VLF propagation in the Earth-ionosphere waveguide

Worldwide lightning location (WWLL) using only 30 lightning sensors has been successfully achieved by using only VLF propagation in the Earth-ionosphere waveguide (EIWG). Ground propagation or mixed "sky" and ground propagation is avoided by requiring evidence of Earth-ionosphere waveguide dispersion. A further requirement is that the lightning strike must be inside the perimeter defined by the lightning sensor sites detecting the stroke. Under these conditions, the time and the location of the stroke can be determined, along with the rms errors. Lightning strokes with errors exceeding 30 Ps or To assist with identifying impulses from the same lightning stroke, the lightning sensor threshold is automatically adjusted to allow an average detection rate of three per second. This largely limits detection to the strongest 4% of all lightning strokes, of which about 40% meet the accuracy requirements for time and location

Modeling of laser keyhole welding: Part I. Mathematical modeling, numerical methodology, role of recoil pressure, multiple reflections, and free surface evolution

A three-dimensional laser-keyhole welding model is developed, featuring the self-consistent evolution of the liquid/vapor (L/V) interface together with full simulation of fluid flow and heat transfer. Important interfacial phenomena, such as free surface evolution, evaporation, kinetic Knudsen layer, homogeneous boiling, and multiple reflections, are considered and applied to the model. The level set approach is adopted to incorporate the L/V interface boundary conditions in the Navier-Stokes equation and energy equation. Both thermocapillary force and recoil pressure, which are the major driving forces for the melt flow, are incorporated in the formulation. For melting and solidification processes at the solid/liquid (S/L) interface, the mixture continuum model has been employed. The article consists of two parts. This article (Part I) presents the model formulation and discusses the effects of evaporation, free surface evolution, and multiple reflections on a steady molten pool to demonstrate the relevance of these interfacial phenomena. The results of the full keyhole simulation and the experimental verification will be provided in the companion article (Part II).close698