Potential role of protease-antiprotease interactions in Perkinsus marinus infection in Crassostrea spp.

Perkinsus marinus causes devastating losses in populations of the eastern oyster (Crassostrea virginica). Our studies have demonstrated that P. marinus secretes extracellular serine proteases which enhance parasite propagation and compromise host defences. Crassostrea virginica. however, possesses several inhibitors of these proteases. The Pacific oyster (C. gigas) is resistant to P. marinus and possesses protease inhibitors with significantly higher specific activities than those in C. virginica. Interestingly, Crassostrea spp. themselves, elaborate metalloprotease activities which can be detected in their plasma, and are increased during P. marinus infections. Together our work suggests that there may be a broad spectrum of humoral host defences that is brought to bear on P. marinus infections by these two Crassostrea species

Mammoth interatrial septal aneurysm in the ICE age

<p>Abstract</p> <p>Background</p> <p>Intracardiac echocardiography (ICE) is a useful imaging modality that is now being used more widely to assist in the percutaneous closure of atrial septal defects (ASD) and patent foramen ovales (PFO).</p> <p>Case presentation</p> <p>A 42 year old lady with a history of transient ischaemic attacks and migraine underwent percutaneous closure of an ASD. Intraprocedural ICE demonstrated a mammoth billowing multiperforated interatrial septal aneurysm in association with a secondum ASD.</p> <p>Conclusion</p> <p>ICE provides excellent adjuvant imaging during percutaneous closure of intracardiac shunts, in this case demonstrating a 'mammoth' interatrial septal aneurysm.</p

Genomic and experimental evidence for multiple metabolic functions in the RidA/YjgF/YER057c/UK114 (Rid) protein family.

BackgroundIt is now recognized that enzymatic or chemical side-reactions can convert normal metabolites to useless or toxic ones and that a suite of enzymes exists to mitigate such metabolite damage. Examples are the reactive imine/enamine intermediates produced by threonine dehydratase, which damage the pyridoxal 5'-phosphate cofactor of various enzymes causing inactivation. This damage is pre-empted by RidA proteins, which hydrolyze the imines before they do harm. RidA proteins belong to the YjgF/YER057c/UK114 family (here renamed the Rid family). Most other members of this diverse and ubiquitous family lack defined functions.ResultsPhylogenetic analysis divided the Rid family into a widely distributed, apparently archetypal RidA subfamily and seven other subfamilies (Rid1 to Rid7) that are largely confined to bacteria and often co-occur in the same organism with RidA and each other. The Rid1 to Rid3 subfamilies, but not the Rid4 to Rid7 subfamilies, have a conserved arginine residue that, in RidA proteins, is essential for imine-hydrolyzing activity. Analysis of the chromosomal context of bacterial RidA genes revealed clustering with genes for threonine dehydratase and other pyridoxal 5'-phosphate-dependent enzymes, which fits with the known RidA imine hydrolase activity. Clustering was also evident between Rid family genes and genes specifying FAD-dependent amine oxidases or enzymes of carbamoyl phosphate metabolism. Biochemical assays showed that Salmonella enterica RidA and Rid2, but not Rid7, can hydrolyze imines generated by amino acid oxidase. Genetic tests indicated that carbamoyl phosphate overproduction is toxic to S. enterica cells lacking RidA, and metabolomic profiling of Rid knockout strains showed ten-fold accumulation of the carbamoyl phosphate-related metabolite dihydroorotate.ConclusionsLike the archetypal RidA subfamily, the Rid2, and probably the Rid1 and Rid3 subfamilies, have imine-hydrolyzing activity and can pre-empt damage from imines formed by amine oxidases as well as by pyridoxal 5'-phosphate enzymes. The RidA subfamily has an additional damage pre-emption role in carbamoyl phosphate metabolism that has yet to be biochemically defined. Finally, the Rid4 to Rid7 subfamilies appear not to hydrolyze imines and thus remain mysterious

Differential Regulation of Extracellular Matrix and Soluble Fibulin-1 Levels by TGF-β<inf>1</inf> in Airway Smooth Muscle Cells

Fibulin-1 (FBLN-1) is a secreted glycoprotein that is associated with extracellular matrix (ECM) formation and rebuilding. Abnormal and exaggerated deposition of ECM proteins is a hallmark of many fibrotic diseases, such as chronic obstructive pulmonary disease (COPD) where small airway fibrosis occurs. The aim of this study was to investigate the regulation of FBLN-1 by transforming growth factor beta 1 (TGF-β1) (a pro-fibrotic stimulus) in primary human airway smooth muscle (ASM) cells from volunteers with and without COPD. Human ASM cells were seeded at a density of 1×104 cells/cm2, and stimulated with or without TGF-β1 (10 ng/ml) for 72 hours before FBLN-1 deposition and soluble FBLN-1 were measured. Fold change in FBLN-1 mRNA was measured at 4, 8, 24, 48, 72 hours. In some experiments, cycloheximide (0.5 μg/ml) was used to assess the regulation of FBLN-1 production. TGF-β1 decreased the amount of soluble FBLN-1 both from COPD and non-COPD ASM cells. In contrast, the deposition of FBLN-1 into the ECM was increased in ASM cells obtained from both groups. TGF-β1 did not increase FBLN-1 gene expression at any of the time points. There were no differences in the TGF-β1 induced FBLN-1 levels between cells from people with or without COPD. Cycloheximide treatment, which inhibits protein synthesis, decreased both the constitutive release of soluble FBLN-1, and TGF-β1 induced ECM FBLN-1 deposition. Furthermore, in cycloheximide treated cells addition of soluble FBLN-1 resulted in incorporation of FBLN-1 into the ECM. Therefore the increased deposition of FBLN-1 by ASM cells into the ECM following treatment with TGF-β1 is likely due to incorporation of soluble FBLN-1 rather than de-novo synthesis. © 2013 Chen et al

Doxycycline reduces the migration of tuberous sclerosis complex-2 null cells - effects on RhoA-GTPase and focal adhesion kinase

& Sons Ltd and Foundation for Cellular and Molecular Medicine. Lymphangioleiomyomatosis (LAM) is associated with dysfunction of the tuberous sclerosis complex (TSC) leading to enhanced cell proliferation and migration. This study aims to examine whether doxycycline, a tetracycline antibiotic, can inhibit the enhanced migration of TSC2-deficient cells, identify signalling pathways through which doxycycline works and to assess the effectiveness of combining doxycycline with rapamycin (mammalian target of rapamycin complex 1 inhibitor) in controlling cell migration, proliferation and wound closure. TSC2-positive and TSC2-negative mouse embryonic fibroblasts (MEF), 323-TSC2-positive and 323-TSC2-null MEF and Eker rat uterine leiomyoma (ELT3) cells were treated with doxycycline or rapamycin alone, or in combination. Migration, wound closure and proliferation were assessed using a transwell migration assay, time-lapse microscopy and manual cell counts respectively. RhoA-GTPase activity, phosphorylation of p70S6 kinase (p70S6K) and focal adhesion kinase (FAK) in TSC2-negative MEF treated with doxycycline were examined using ELISA and immunoblotting techniques. The enhanced migration of TSC2-null cells was reduced by doxycycline at concentrations as low as 20 pM, while the rate of wound closure was reduced at 2-59 μM. Doxycycline decreased RhoA-GTPase activity and phosphorylation of FAK in these cells but had no effect on the phosphorylation of p70S6K, ERK1/2 or AKT. Combining doxycycline with rapamycin significantly reduced the rate of wound closure at lower concentrations than achieved with either drug alone. This study shows that doxycycline inhibits TSC2-null cell migration. Thus doxycycline has potential as an anti-migratory agent in the treatment of diseases with TSC2 dysfunction. © 2015 John Wile

Hybrid active focusing with adaptive dispersion for higher defect sensitivity in guided wave inspection of cylindrical structures

This is an Accepted Manuscript of an article published by Taylor & Francis in Nondestructive Testing and Evaluation on 23/11/2015, available online: https://www.tandfonline.com/doi/full/10.1080/10589759.2015.1093628.Ultrasonic guided wave inspection is widely used for scanning prismatic structures such as pipes for metal loss. Recent research has investigated focusing the sound energy into predetermined regions of a pipe in order to enhance the defect sensitivity. This paper presents an active focusing technique which is based on a combination of numerical simulation and time reversal concept. The proposed technique is empirically validated using a 3D laser vibrometry measurement of the focal spot. The defect sensitivity of the proposed technique is compared with conventional active focusing, time reversal focusing and synthetic focusing through an empirically validated finite element parametric study. Based on the results, the proposed technique achieves approximately 10 dB improvement of signal-to-coherent-noise ratio compared to the conventional active focusing and time reversal focusing. It is also demonstrated that the proposed technique to have an amplitude gain of around 5 dB over synthetic focusing for defects <0.5λs. The proposed technique is shown to have the potential to improve the reliably detectable flaw size in guided wave inspection from 9% to less than 1% cross-sectional area loss.TWI Ltd and the Center for Electronic System Research (CESR) of Brunel University

Hybrid active focusing with adaptive dispersion for higher defect sensitivity in guided wave inspection of cylindrical structures

This is an Accepted Manuscript of an article published by Taylor & Francis in Nondestructive Testing and Evaluation on 23/11/2015, available online: https://www.tandfonline.com/doi/full/10.1080/10589759.2015.1093628.Ultrasonic guided wave inspection is widely used for scanning prismatic structures such as pipes for metal loss. Recent research has investigated focusing the sound energy into predetermined regions of a pipe in order to enhance the defect sensitivity. This paper presents an active focusing technique which is based on a combination of numerical simulation and time reversal concept. The proposed technique is empirically validated using a 3D laser vibrometry measurement of the focal spot. The defect sensitivity of the proposed technique is compared with conventional active focusing, time reversal focusing and synthetic focusing through an empirically validated finite element parametric study. Based on the results, the proposed technique achieves approximately 10 dB improvement of signal-to-coherent-noise ratio compared to the conventional active focusing and time reversal focusing. It is also demonstrated that the proposed technique to have an amplitude gain of around 5 dB over synthetic focusing for defects <0.5λs. The proposed technique is shown to have the potential to improve the reliably detectable flaw size in guided wave inspection from 9% to less than 1% cross-sectional area loss.TWI Ltd and the Center for Electronic System Research (CESR) of Brunel University

LF-15 &amp; T7, synthetic peptides derived from tumstatin, attenuate aspects of airway remodelling in a murine model of chronic OVA-induced allergic airway disease

Background: Tumstatin is a segment of the collagen-IV protein that is markedly reduced in the airways of asthmatics. Tumstatin can play an important role in the development of airway remodelling associated with asthma due to its antiangiogenic properties. This study assessed the anti-angiogenic properties of smaller peptides derived from tumstatin, which contain the interface tumstatin uses to interact with the aVb3 integrin. Methods: Primary human lung endothelial cells were exposed to the LF-15, T3 and T7 tumstatin-derived peptides and assessed for cell viability and tube formation in vitro. The impact of the anti-angiogenic properties on airways hyperresponsiveness (AHR) was then examined using a murine model of chronic OVA-induced allergic airways disease. Results: The LF-15 and T7 peptides significantly reduced endothelial cell viability and attenuated tube formation in vitro. Mice exposed to OVA+ LF-15 or OVA+T7 also had reduced total lung vascularity and AHR was attenuated compared to mice exposed to OVA alone. T3 peptides reduced cell viability but had no effect on any other parameters. Conclusion: The LF-15 and T7 peptides may be appropriate candidates for use as novel pharmacotherapies due to their small size and anti-angiogenic properties observed in vitro and in vivo. © 2014 Grafton et al

MC-Simulation of the Transverse Double Spin Asymmetry for RHIC

Using {\sc Sphinx tt}, a new MC simulation program for transverse polarized nucleon--nucleon scattering based on {\sc Pythia~5.6}, we calculate the transverse double spin asymmetry $A^{TT}$ in the Drell-Yan process. If one assumes (quite arbitrarily) that the transversity parton distribution $\delta q(x,Q^2)$ equals the helicity distribution $\Delta q(x,Q^2)$ at some low $Q_0^2$ scale, the resulting asymmetry is of order 1\%. In this case is $A^{TT}$ would hardly be be measurable with PHENIX at RHIC.Comment: 17 pages, 5 figure

Differential deposition of fibronectin by asthmatic bronchial epithelial cells

© 2015 the American Physiological Society. Altered ECM protein deposition is a feature in asthmatic airways. Fibronectin (Fn), an ECM protein produced by human bronchial epithelial cells (HBECs), is increased in asthmatic airways. This study investigated the regulation of Fn production in asthmatic or nonasthmatic HBECs and whether Fn modulated HBEC proliferation and inflammatory mediator secretion. The signaling pathways underlying transforming growth factor (TGF)-β1-regulated Fn production were examined using specific inhibitors for ERK, JNK, p38 MAPK, phosphatidylinositol 3 kinase, and activin-like kinase 5 (ALK5). Asthmatic HBECs deposited higher levels of Fn in the ECM than nonasthmatic cells under basal conditions, whereas cells from the two groups had similar levels of Fn mRNA and soluble Fn. TGF-β1 increased mRNA levels and ECM and soluble forms of Fn but decreased cell proliferation in both cells. The rate of increase in Fn mRNA was higher in nonasthmatic cells. However, the excessive amounts of ECM Fn deposited by asthmatic cells after TGF-β1 stimulation persisted compared with nonasthmatic cells. Inhibition of ALK5 completely prevented TGF- β1-induced Fn deposition. Importantly, ECM Fn increased HBEC proliferation and IL-6 release, decreased PGE2 secretion, but had no effect on VEGF release. Soluble Fn had no effect on cell proliferation and inflammatory mediator release. Asthmatic HBECs are intrinsically primed to produce more ECM Fn, which when deposited into the ECM, is capable of driving remodeling and inflammation. The increased airway Fn may be one of the key driving factors in the persistence of asthma and represents a novel, therapeutic target