Extracellular ATP induces fast and transient non-selective cationic currents and cytosolic Ca2+ changes in human umbilical artery smooth muscle cells

Ionotropic purinergic receptors (P2X) are expressed in endothelial and smooth muscle cells of blood vessels. ATP acting on smooth muscle P2X receptors is able to induce vasoconstriction in different kind of vessels. However, to our knowledge, there are no reports that directly show the activity of these purinergic receptors in native human vascular smooth muscle cells. In this work, we describe for the first time an ATP-induced current in freshly isolated human umbilical artery (HUA) smooth muscle cells. The current was measured by patch-clamp technique in whole-cell condition on cells clamped at −50 mV. At 100 μM of ATP the current showed a rapid activation and desensitization, and was carried by both Na+ and Ca2+. The current was completely blocked by suramin (300 μM) and partially blocked by 100 μM of Zn2+ without affecting the kinetic of desensitization. All these properties suggest that the ATP-induced ionic currents are mediated through P2X1-like receptors. Moreover, we show that ATP transiently increased cytosolic Ca2+ in “in situ” smooth muscle cells of intact HUA segments and that this response is dependent of extracellular and intracellular Ca2+. These data expand the knowledge of purinergic receptors properties in vascular smooth muscle cells and the probable role of ATP as a paracrine modulator of contractile tone in a human artery which is fundamental for feto-placental blood flow

Improving clinical paediatric research and learning from COVID-19: recommendations by the Conect4Children expert advice group

none42Background: The COVID-19 pandemic has had a devastating impact on multiple aspects of healthcare, but has also triggered new ways of working, stimulated novel approaches in clinical research and reinforced the value of previous innovations. Conect4children (c4c, www.conect4children.org) is a large collaborative European network to facilitate the development of new medicines for paediatric populations, and is made up of 35 academic and 10 industry partners from 20 European countries, more than 50 third parties, and around 500 affiliated partners. Methods: We summarise aspects of clinical research in paediatrics stimulated and reinforced by COVID-19 that the Conect4children group recommends regulators, sponsors, and investigators retain for the future, to enhance the efficiency, reduce the cost and burden of medicines and non-interventional studies, and deliver research-equity. Findings: We summarise aspects of clinical research in paediatrics stimulated and reinforced by COVID-19 that the Conect4children group recommends regulators, sponsors, and investigators retain for the future, to enhance the efficiency, reduce the cost and burden of medicines and non-interventional studies, and deliver research-equityWe provide examples of research innovation, and follow this with recommendations to improve the efficiency of future trials, drawing on industry perspectives, regulatory considerations, infrastructure requirements and parent–patient–public involvement. We end with a comment on progress made towards greater international harmonisation of paediatric research and how lessons learned from COVID-19 studies might assist in further improvements in this important area.noneRamanan A.V.; Modi N.; de Wildt S.N.; Aurich B.; Bakhtadze S.; Sirvent F.J.B.; Cabanas F.; Campbell L.; Casanova M.; Charlton P.; Crandall W.; Eichler I.; Fregonese L.; Hawcutt D.B.; Iveli P.; Jaki T.; Jocic-Jakubi B.; Johnson M.; Kaguelidou F.; Karadag B.; Kelly L.E.; Lim M.; Moreno C.; Neumann E.; Ollivier C.; Oualha M.; Raffaeli G.; Ribeiro M.A.; Roilides E.; de Rojas T.; Simon A.R.S.; Ruperto N.; Scarpa M.; Schwab M.; Siapkara A.; Singh Y.; Smits A.; Striano P.; Urru S.A.M.; Vivarelli M.; de Wildt S.; Zivkoviz Z.Ramanan, A. V.; Modi, N.; de Wildt, S. N.; Aurich, B.; Bakhtadze, S.; Sirvent, F. J. B.; Cabanas, F.; Campbell, L.; Casanova, M.; Charlton, P.; Crandall, W.; Eichler, I.; Fregonese, L.; Hawcutt, D. B.; Iveli, P.; Jaki, T.; Jocic-Jakubi, B.; Johnson, M.; Kaguelidou, F.; Karadag, B.; Kelly, L. E.; Lim, M.; Moreno, C.; Neumann, E.; Ollivier, C.; Oualha, M.; Raffaeli, G.; Ribeiro, M. A.; Roilides, E.; de Rojas, T.; Simon, A. R. S.; Ruperto, N.; Scarpa, M.; Schwab, M.; Siapkara, A.; Singh, Y.; Smits, A.; Striano, P.; Urru, S. A. M.; Vivarelli, M.; de Wildt, S.; Zivkoviz, Z

Development of a neonatal adverse event severity scale through a Delphi consensus approach

Background: Assessment of the seriousness, expectedness and causality are necessary for any adverse event (AE) in a clinical trial. In addition, assessing AE severity helps determine the importance of the AE in the clinical setting. Standardisation of AE severity criteria could make safety information more reliable and comparable across trials. Although standardised AE severity scales have been developed in other research fields, they are not suitable for use in neonates. The development of an AE severity scale to facilitate the conduct and interpretation of neonatal clinical trials is therefore urgently needed. / Methods: A stepwise consensus process was undertaken within the International Neonatal Consortium (INC) with input from all relevant stakeholders. The consensus process included several rounds of surveys (based on a Delphi approach), face-to-face meetings and a pilot validation. / Results: Neonatal AE severity was classified by five grades (mild, moderate, severe, life threatening or death). AE severity in neonates was defined by the effect of the AE on age appropriate behaviour, basal physiological functions and care changes in response to the AE. Pilot validation of the generic criteria revealed κ=0.23 and guided further refinement. This generic scale was applied to 35 typical and common neonatal AEs resulting in the INC neonatal AE severity scale (NAESS) V.1.0, which is now publicly available. / Discussion: The INC NAESS is an ongoing effort that will be continuously updated. Future perspectives include further validation and the development of a training module for users