Intracellular pathways of calcitonin gene‐related peptide‐induced relaxation of human coronary arteries:A key role for Gβγ subunit instead of cAMP

Background and Purpose: Calcitonin gene‐related peptide (CGRP) is a potent vasodilator. While its signalling is assumed to be mediated via increases in cAMP, this study focused on elucidating the actual intracellular signalling pathways involved in CGRP‐induced relaxation of human isolated coronary arteries (HCA). Experimental Approach: HCA were obtained from heart valve donors (27 M, 25 F, age 54 ± 2 years). Concentration–response curves to human α‐CGRP or forskolin were constructed in HCA segments, incubated with different inhibitors of intracellular signalling pathways, and intracellular cAMP levels were measured with and without stimulation. Results: Adenylyl cyclase (AC) inhibitors SQ22536 + DDA and MDL‐12330A, and PKA inhibitors Rp‐8‐Br‐cAMPs and H89, did not inhibit CGRP‐induced relaxation of HCA, nor did the guanylyl cyclase inhibitor ODQ, PKG inhibitor KT5823, EPAC1/2 inhibitor ESI09, potassium channel blockers TRAM‐34 + apamin, iberiotoxin or glibenclamide, or the Gαq inhibitor YM‐254890. Phosphodiesterase inhibitors induced a concentration‐dependent decrease in the response to KCl but did not potentiate relaxation to CGRP. Relaxation to forskolin was not blocked by PKA or AC inhibitors, although AC inhibitors significantly inhibited the increase in cAMP. Inhibition of Gβγ subunits using gallein significantly inhibited the relaxation to CGRP in human coronary arteries. Conclusion: While CGRP signalling is generally assumed to act via cAMP, the CGRP‐induced vasodilation in HCA was not inhibited by targeting this intracellular signalling pathway at different levels. Instead, inhibition of Gβγ subunits did inhibit the relaxation to CGRP, suggesting a different mechanism of CGRP‐induced relaxation than generally believed

25th annual computational neuroscience meeting: CNS-2016

The same neuron may play different functional roles in the neural circuits to which it belongs. For example, neurons in the Tritonia pedal ganglia may participate in variable phases of the swim motor rhythms [1]. While such neuronal functional variability is likely to play a major role the delivery of the functionality of neural systems, it is difficult to study it in most nervous systems. We work on the pyloric rhythm network of the crustacean stomatogastric ganglion (STG) [2]. Typically network models of the STG treat neurons of the same functional type as a single model neuron (e.g. PD neurons), assuming the same conductance parameters for these neurons and implying their synchronous firing [3, 4]. However, simultaneous recording of PD neurons shows differences between the timings of spikes of these neurons. This may indicate functional variability of these neurons. Here we modelled separately the two PD neurons of the STG in a multi-neuron model of the pyloric network. Our neuron models comply with known correlations between conductance parameters of ionic currents. Our results reproduce the experimental finding of increasing spike time distance between spikes originating from the two model PD neurons during their synchronised burst phase. The PD neuron with the larger calcium conductance generates its spikes before the other PD neuron. Larger potassium conductance values in the follower neuron imply longer delays between spikes, see Fig. 17.Neuromodulators change the conductance parameters of neurons and maintain the ratios of these parameters [5]. Our results show that such changes may shift the individual contribution of two PD neurons to the PD-phase of the pyloric rhythm altering their functionality within this rhythm. Our work paves the way towards an accessible experimental and computational framework for the analysis of the mechanisms and impact of functional variability of neurons within the neural circuits to which they belong

Van co-design tot playtest: een leidraad voor een player-centered design process

Het e-Treasure consortium heeft de afgelopen twee jaar een proces verfijnd dat spelontwikkelaars en e-learning bedrijven kan helpen met de sprong naar Meaningful Play. Het handboek is als volgt opgebouwd. In het eerste hoofdstuk overlopen we wat we bedoelen met Meaningful Play en de noodzaak voor een Player- Centered Design proces. In het tweede hoofdstuk geven we theoretische achtergrondinformatie. De User-Centered design cyclus”, waarvan de Player-Centered Design cyclus uiteraard een specifieke toepassing hoort te zijn, wordt allereerst samenvattend beschreven met uitleg en achtergrond bij de diverse stappen. Vervolgens wordt dieper ingegaan op een aantal methodologische aspecten, waarbij voor de verschillende methodologieën specifieke voor- en nadelen worden aangehaald, en argumenten waarom men (niet) voor een bepaalde methode zou kiezen. In het derde en laatste hoofdstuk illustreren we aan de hand van het e- Treasure project hoe we dit proces en de technieken hebben toegepast. We geven ook anekdotes, problemen en hoe dit op te lossen.nrpages: 35status: publishe

Pneumococcal aetiology and serotype distribution in paediatric community-acquired pneumonia.

Community-acquired pneumonia (CAP) is a major cause of morbidity in children. This study estimated the proportion of children with pneumococcal CAP among children hospitalised with CAP in Belgium and describes the causative serotype distribution after implementation of the 7-valent pneumococcal conjugate vaccine. Children 0-14 years hospitalised with X-ray-confirmed CAP were prospectively enrolled in a multicentre observational study. Acute and convalescent blood samples were collected. Pneumococcal aetiology was assessed by conventional methods (blood or pleural fluid cultures with Quellung reaction capsular typing or polymerase chain reaction [PCR] in pleural fluid), and recently developed methods (real-time PCR in blood and World Health Organization-validated serotype-specific serology). A total of 561 children were enrolled. Pneumococcal aetiology was assessed by conventional methods in 539, serology in 171, and real-time PCR in blood in 154. Pneumococcal aetiology was identified in 12.2% (66/539) of the children by conventional methods alone but in 73.9% by the combination of conventional and recently developed methods. The pneumococcal detection rate adjusted for the whole study population was 61.7%. Serotypes 1 (42.3%), 5 (16.0%), and 7F(7A) (12.8%) were predominant. In conclusion, Streptococcus pneumoniae remains the predominant bacteria in children hospitalised for CAP in Belgium after implementation of 7-valent pneumococcal conjugate vaccine, with non-vaccine-serotypes accounting for the majority of cases. The use of recently developed methods improves diagnosis of pneumococcal aetiology

Implantace energetických Au iontů do ZnO nanopilířů pro modulaci optické odezvy

Nanopillars of ZnO were implanted with Au-400 keV ions at various ion fluences from 1 x 10(15) cm(-2) to 1 x 10(16) cm(-2) and subsequently annealed at 750 degrees C for 15 min in order to reduce the implantation damage and to support Au nanoparticle (NP) aggregation. It was found that implantation-induced effects and thermal effects influence the Au NP coalescence as well as the quality of the ZnO nanopillars. Rutherford Back-Scattering spectrometry (RBS) showed the broader Au-depth profiles than it was theoretically predicted, but the Au-concentration maximum agrees well with prediction taking into account the effective ZnO layer density. The implantation at the higher fluences induced the morphology modification of the nanopillar layer evidenced by RBS and scanning electron microscopy (SEM). An indirect evidence of this effect was given by optical ellipsometry due to gradual refractive index changes in the ZnO nanopillars with the increased Au-ion fluence. Optical characterization of the Au-implanted and annealed nanopillars performed by means of photoluminescence (PL) and diffuse-reflectance spectroscopy (DRS) evidenced the surface plasmon resonance (SPR) activity of the embedded Au NPs. The SPR-enhanced scattering and PL emission observed in the spectral range 500-650 nm are ascribed to Au NPs or more complex Au-clusters. In addition, the ellipsometry measurements of extinction coefficient are found to corroborate well results from DRS, both indicating increase of SPR effect with the increase of Au-ion fluence and after the post-annealing.Do nanopilířů ZnO byly implantovány ionty Au-400 keV při různých fluktuacích iontů od 1 x 10(15) cm(-2) do 1 x 10(16) cm(-2) a následně žíhány při 750 stupních C po dobu 15 min, aby se snížilo poškození implantace a podpořila se agregace nanočástic Au (NP). Bylo zjištěno, že efekty vyvolané implantací a tepelné efekty ovlivňují koalescenci Au NP a také kvalitu nanopilířů ZnO. Rutherfordova zpětná rozptylová spektrometrie (RBS) ukázala širší profily hloubky Au, než bylo teoreticky předpovězeno, ale maximum koncentrace Au dobře souhlasí s predikcí, která bere v úvahu efektivní hustotu vrstvy ZnO. Implantace při vyšších fluencesách vyvolala morfologickou modifikaci nanopilární vrstvy doloženou RBS a rastrovací elektronovou mikroskopií (SEM). Nepřímý důkaz tohoto efektu poskytla optická elipsometrie v důsledku postupných změn indexu lomu v nanopilárech ZnO se zvýšeným fluencem Au-iontů. Optická charakterizace Au-implantovaných a žíhaných nanopilárů provedená pomocí fotoluminiscence (PL) a difuzně-reflexní spektroskopie (DRS) prokázala aktivitu povrchové plasmonové rezonance (SPR) vložených Au NP. SPR-enhanced rozptyl a PL emise pozorované ve spektrálním rozsahu 500-650 nm jsou připisovány Au NP nebo složitějším Au-klastrům. Kromě toho bylo zjištěno, že elipsometrická měření extinkčního koeficientu dobře potvrzují výsledky z DRS, což ukazuje na zvýšení účinku SPR se zvýšením fluence Au-iontů a po následném žíhání

Original Article Simulation and Robotics Studies of Salamander Locomotion Applying Neurobiological Principles to the Control of Locomotion in Robots

This article presents a project that aims at understanding the neural circuitry controlling salamander locomotion, and developing an amphibious salamander-like robot capable of replicating its bimodal locomotion, namely swimming and terrestrial walking. The controllers of the robot are central pattern generator models inspired by the salamander’s locomotion control network. The goal of the project is twofold: (1) to use robots as tools for gaining a better understanding of locomotion control in vertebrates and (2) to develop new robot and control technologies for developing agile an

P-III: A Player-Centered, Iterative, Interdisciplinary and Integrated Framework for Serious Game Design and Development

While reconciling a creative game design process with a complex software engineering process is already a daunting task, serious games add another ingredient to an already volatile mixture: the challenge of crafting an effective learning experience. In order to achieve this strenuous objective, Group T's e-Media Lab and the Centre for User Experience Research, K.U.Leuven, have developed a player-centered, iterative, interdisciplinary and integrated (P-III) framework. This framework has been developed over the course of five years of research on the design and development of serious games. Hence, P-III is built bottom-up, molded and shaped, tested and refined through several research projects [1,9,17,18,19,20,21,22,23]. While P-III also prescribes a specific process, in this paper we limit ourselves to highlighting the four pillars of the P-III framework, and their theoretical underpinnings. © 2012 Springer-Verlag.status: publishe

Intracellular pathways of calcitonin gene-related peptide-induced relaxation of human coronary arteries: A key role for Gβγ subunit instead of cAMP.

Publication status: PublishedBACKGROUND AND PURPOSE: Calcitonin gene-related peptide (CGRP) is a potent vasodilator. While its signalling is assumed to be mediated via increases in cAMP, this study focused on elucidating the actual intracellular signalling pathways involved in CGRP-induced relaxation of human isolated coronary arteries (HCA). EXPERIMENTAL APPROACH: HCA were obtained from heart valve donors (27 M, 25 F, age 54 ± 2 years). Concentration-response curves to human α-CGRP or forskolin were constructed in HCA segments, incubated with different inhibitors of intracellular signalling pathways, and intracellular cAMP levels were measured with and without stimulation. RESULTS: Adenylyl cyclase (AC) inhibitors SQ22536 + DDA and MDL-12330A, and PKA inhibitors Rp-8-Br-cAMPs and H89, did not inhibit CGRP-induced relaxation of HCA, nor did the guanylyl cyclase inhibitor ODQ, PKG inhibitor KT5823, EPAC1/2 inhibitor ESI09, potassium channel blockers TRAM-34 + apamin, iberiotoxin or glibenclamide, or the Gαq inhibitor YM-254890. Phosphodiesterase inhibitors induced a concentration-dependent decrease in the response to KCl but did not potentiate relaxation to CGRP. Relaxation to forskolin was not blocked by PKA or AC inhibitors, although AC inhibitors significantly inhibited the increase in cAMP. Inhibition of Gβγ subunits using gallein significantly inhibited the relaxation to CGRP in human coronary arteries. CONCLUSION: While CGRP signalling is generally assumed to act via cAMP, the CGRP-induced vasodilation in HCA was not inhibited by targeting this intracellular signalling pathway at different levels. Instead, inhibition of Gβγ subunits did inhibit the relaxation to CGRP, suggesting a different mechanism of CGRP-induced relaxation than generally believed

Comparison of SSS and rtPCR in blood results for the 126 patients evaluated by both additional methods.

<p>Values are numbers (%) of subjects. P-CAP, pneumococcal community-acquired pneumonia; PCR, polymerase chain reaction; rtPCR, real-time polymerase chain reaction; SSS, serotype-specific serology.</p>a<p>SSS included serotypes 1, 5, 6B(6D), 7F(7A), 9V(9N), 14, 19A, 19F, and 23F.</p>b<p>rtPCR in blood included serotypes 1, 3, 4, 5, 6A(6C), 6B(6D), 7F(7A), 9V(9N), 14, 18C(18B), 19A, 19F, and 23F.</p