Suppression of Adenosine-Activated Chloride Transport by Ethanol in Airway Epithelia

Alcohol abuse is associated with increased lung infections. Molecular understanding of the underlying mechanisms is not complete. Airway epithelial ion transport regulates the homeostasis of airway surface liquid, essential for airway mucosal immunity and lung host defense. Here, air-liquid interface cultures of Calu-3 epithelial cells were basolaterally exposed to physiologically relevant concentrations of ethanol (0, 25, 50 and 100 mM) for 24 hours and adenosine-stimulated ion transport was measured by Ussing chamber. The ethanol exposure reduced the epithelial short-circuit currents (ISC) in a dose-dependent manner. The ion currents activated by adenosine were chloride conductance mediated by cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated chloride channel. Alloxazine, a specific inhibitor for A2B adenosine receptor (A2BAR), largely abolished the adenosine-stimulated chloride transport, suggesting that A2BAR is a major receptor responsible for regulating the chloride transport of the cells. Ethanol significantly reduced intracellular cAMP production upon adenosine stimulation. Moreover, ethanol-suppression of the chloride secretion was able to be restored by cAMP analogs or by inhibitors to block cAMP degradation. These results imply that ethanol exposure dysregulates CFTR-mediated chloride transport in airways by suppression of adenosine-A2BAR-cAMP signaling pathway, which might contribute to alcohol-associated lung infections

Wave interaction with defects in pressurised composite structures

There exists a great variety of structural failure modes which must be frequently inspected to ensure continuous structural integrity of composite structures. This work presents a Finite Element (FE) based method for calculating wave interaction with damage within structures of arbitrary layering and geometric complexity. The principal novelty is the investigation of pre-stress effect on wave propagation and scattering in layered structures. A Wave Finite Element (WFE) method, which combines FE analysis with periodic structure theory (PST), is used to predict the wave propagation properties along periodic waveguides of the structural system. This is then coupled to the full FE model of a coupling joint within which structural damage is modelled, in order to quantify wave interaction coeffcients through the joint. Pre-stress impact is quantified by comparison of results under pressurised and non-pressurised scenarios. The results show that including these pressurisation effects in calculations is essential. This is of specific relevance to aircraft structures being intensely pressurised while on air. Numerical case studies are exhibited for different forms of damage type. The exhibited results are validated against available analytical and experimental results

Thiram inhibits angiogenesis and slows the development of experimental tumours in mice

Thiram-tetramethylthiuram disulphide – a chelator of heavy metals, inhibited DNA synthesis and induced apoptosis in cultured bovine capillary endothelial cells. Bovine capillary endothelial cells were 10–60-fold more sensitive to thiram than other cell types. These effects were prevented by addition of antioxidants, indicating involvement of reactive oxygen species. Exogenously added Cu2+ impeded specifically and almost completely the inhibitory effect of thiram for bovine capillary endothelial cells. Moreover, thiram had markedly inhibited human recombinant Cu/Zn superoxide dismutase enzymatic activity (85%) in vitro. Moreover, PC12-SOD cells with elevated Cu/Zn superoxide dismutase were less sensitive to thiram treatment than control cells. These data indicate that the effects of thiram are mediated by inhibition of Cu/Zn superoxide dismutase activity. Oral administration of thiram (13–30 μg mouse−1), inhibited angiogenesis in CD1 nude mice. Tumour development is known to largely depend on angiogenesis. We found that oral administration of thiram (30 μg) to mice caused significant inhibition of C6 glioma tumour development (60%) and marked reduction (by 3–5-fold) in metastatic growth of Lewis lung carcinoma. The data establish thiram as a potential inhibitor of angiogenesis and raise the possibility for its use as therapy in pathologies in which neovascularisation is involved, including neoplasia

Roflumilast partially reverses smoke-induced mucociliary dysfunction

BACKGROUND: Phosphodiesterases (PDEs) break down cAMP, thereby regulating intracellular cAMP concentrations and diffusion. Since PDE4 predominates in airway epithelial cells, PDE4 inhibitors can stimulate Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) by increasing cAMP. Tobacco smoking and COPD are associated with decreased CFTR function and impaired mucociliary clearance (MCC). However, the effects of the PDE4 inhibitor roflumilast on smoke-induced mucociliary dysfunction have not been fully explored. METHODS: Primary normal human bronchial epithelial cells (NHBE) from non-smokers, cultured at the air-liquid interface (ALI) were used for most experiments. Cultures were exposed to cigarette smoke in a Vitrocell VC-10 smoking robot. To evaluate the effect of roflumilast on intracellular cAMP concentrations, fluorescence resonance energy transfer (FRET) between CFP- and YFP-tagged protein kinase A (PKA) subunits was recorded. Airway surface liquid (ASL) was measured using light refraction scanning and ciliary beat frequency (CBF) employing infrared differential interference contrast microscopy. Chloride conductance was measured in Ussing chambers and CFTR expression was quantified with qPCR. RESULTS: While treatment with 100 nM roflumilast had little effect alone, it increased intracellular cAMP upon stimulation with forskolin and albuterol in cultures exposed to cigarette smoke and in control conditions. cAMP baselines were lower in smoke-exposed cells. Roflumilast prolonged cAMP increases in smoke-exposed and control cultures. Smoke-induced reduction in functional, albuterol-mediated chloride conductance through CFTR was improved by roflumilast. ASL volumes also increased in smoke-exposed cultures in the presence of roflumilast while it did not in its absence. Cigarette smoke exposure decreased CBF, an effect rescued with roflumilast, particularly when used together with the long-acting ß-mimetic formoterol. Roflumilast also enhanced forskolin-induced CBF stimulation in ASL volume supplemented smoked and control cells, confirming the direct stimulatory effect of rising cAMP on ciliary function. In active smokers, CFTR mRNA expression was increased compared to non-smokers and ex-smokers. Roflumilast also increased CFTR mRNA levels in cigarette-smoke exposed cell cultures. CONCLUSIONS: Our results show that roflumilast can rescue smoke-induced mucociliary dysfunction by reversing decreased CFTR activity, augmenting ASL volume, and stimulating CBF, the latter particularly in combination with formoterol. As expected, CFTR mRNA expression was not indicative of apical CFTR function

Bioinformatics and molecular modeling in glycobiology

The field of glycobiology is concerned with the study of the structure, properties, and biological functions of the family of biomolecules called carbohydrates. Bioinformatics for glycobiology is a particularly challenging field, because carbohydrates exhibit a high structural diversity and their chains are often branched. Significant improvements in experimental analytical methods over recent years have led to a tremendous increase in the amount of carbohydrate structure data generated. Consequently, the availability of databases and tools to store, retrieve and analyze these data in an efficient way is of fundamental importance to progress in glycobiology. In this review, the various graphical representations and sequence formats of carbohydrates are introduced, and an overview of newly developed databases, the latest developments in sequence alignment and data mining, and tools to support experimental glycan analysis are presented. Finally, the field of structural glycoinformatics and molecular modeling of carbohydrates, glycoproteins, and protein–carbohydrate interaction are reviewed

A comparative numerical study of standard 3D composite plate using virtual crack closure technique and cohesive zone modelling method

A standard square plate specimen made up of Carbon Fibre Reinforced Polymer (CFRP) and Glass Fibre Reinforced Polymer (GFRP) laminates with an artificially-introduced delamination was considered for evaluation of fracture parameters using Abaqus-Standard software. Though there are several methods to evaluate the initiation of delamination using software, Virtual Crack Closure Technique (VCCT) and Cohesive Zone Modelling (CZM) method are commonly used. By using the above-mentioned methods, the Quasi-Isotropic (QI) composite laminates were analysed for varying compressive loads. The Load-Displacement responses were reported and seen to be in overall good statement with each other, while the modelling effort and computational time required were quite higher in CZM method compared to VCCT. Onset of delamination growth for CFRP and GFRP laminates was studied using VCCT by evaluating Strain Energy Release Rates (SERR). The results for both the specimens were compared and reported

A Comparative Analytical study of Standard Composite Double Cantilever Beam using Virtual Crack Closure Technique and Cohesive Zone Modelling method.

Double cantilever beam specimens made up of Carbon Fibre-Reinforced Polymer laminate (CFRP) and Glass Fibre-Reinforced Polymer (GFRP) laminate were modelled, analysed and their fracture parameters were evaluated using Abaqus-Standard. For the quasi-isotropic specimens, load-displacement responses were computed by varying tensile loads using Virtual Crack Closure Technique (VCCT) and Cohesive Zone Modelling (CZM) method. From the results, it was seenthat modelling effort and computational-time required were quite higher in CZM method. Onset of delamination for both specimens were studied using VCCT after computing energy release rates. Also, the delamination pattern onset was found comparable in both the materials with delay of initiation in GFRP laminate specimen