Development of a HPLC-UV method for the simultaneous determination of intracellular glutathione species in human cells

In the present work, an HPLC-UV method was set-up to allow the simultaneous quantification of the reduced-GSH, oxidised-GSSG and nitroso-GSNO glutathione species. Chromatographic separation was achieved on YMC ODS-A C18 column (150 × 4.6 mm, 5 μm), coupled to a Guard-c precolumn (YMC-Pack, 10 × 1-4,0 mm). The eluted compounds were detected at 215 nm by UV-detector, by keeping the column oven at room temperature while the auto-sampler temperature was maintained at 4°C. A fractional factorial design has been applied for the optimization of the mobile phase resulting in baseline separated peaks within 6 minutes. In-house validation was evaluated by linearity, limits of detection (LODs), limits of quantification (LOQs), reproducibility, repeatability and recovery. The detection and quantification limits obtained for standard solutions were below 0.2 μM and 0.6 μM, respectively (RSD values below 2%). The developed method was applied to the measurement of GSH, GSSG and GSNO in human pulmonary cells (A549) exposed to limonene, limonene oxide solubilized into the culture medium and to NO2 as gas phase. Results show an increase in GSH levels, without significant changes in GSSG, when cells were exposed to limonene oxide, while cells exposed to NO2 resulted in a significant increase of GSNO amount. Detection limits were of 1 μM for the glutathione species measured in A549 cells, with RSD values below 2.5%. In conclusion, the present HPLC-UV method can be readily used to measure in a rapid, simultaneous and accurate way the status of GSH, GSSG and GSNO in human cells, their simultaneous quantification helping to better predict the potential impact of chemicals on human health.JRC.I.1-Chemical Assessment and Testin

Migration of Polycyclic Aromatic Hydrocarbons (PAHs) from plastic and rubber articles

Polycyclic Aromatic Hydrocarbons (PAHs) constitute a large group of chemically related substances many of which are known carcinogens. To minimise human exposure there are already several pieces of EU legislation which limit their presence in certain food products, as well as in water and ambient air. Under the REACH regulation (EC 1907/2006 Annex XVII, Entry 50), eight priority PAHs have for some time been restricted in extender oils used in tyres. Although not added deliberately to consumer products, PAHs can still be present as impurities. An amendment of the above mentioned legislation (Regulation EU 1272/2013) establishes content limits for the eight PAHs of 0.5 mg kg-1 for plastic and rubber components of toys/childcare articles, and 1 mg kg-1 for all other consumer articles, in direct and prolonged, or short-term repetitive, contact with the skin or oral cavity. In May 2016 DG JRC and DG GROW signed an Administrative Arrangement (AA 34003) known as the STANPAHs project. The main objective of this contract was for the JRC to provide scientific support in the implementation and potential amendment of the restriction on polycyclic aromatic hydrocarbons, in particular concerning paragraphs 5 and 6 of entry 50 of Annex XVII to the REACH legislation. The main objectives of the project were: a) to gain a better understanding of the migration behaviour of certain PAHs in plastic and rubber components of articles, and b) to develop a reliable methodology to determine PAH migration from these matrices, under conditions simulating, to the best possible extent, dermal contact (including the oral cavity). This report presents the outcomes of the experimental studies carried out at JRC and the achievements towards fulfilling these objectives. A set of manufactured polymeric plastic and rubber matrices, to be used as test materials in the project, has been chosen based on criteria such as their frequency of use in articles within the scope of the restriction and the likelihood of the presence of high PAH contents (e.g. due to their content in carbon black or extender oils). Various grades and types of ingredients known to be PAH sources were used in the formulation of the manufactured ad-hoc materials. The test materials included low density polyethylene (LDPE), polystyrene (PS) and polyvinyl chloride (PVC) as plastic matrices, and ethylene-propylene diene monomer (EPDM), natural rubber-butadiene rubber (NR-BR) and silicone as rubber matrices. Moreover, recycled granules (coated and uncoated) originating from end-of-life tyres produced before and after 2010 as well as rubber tiles made of the recycled coated granules were also made available for this study. The content of each of the eight restricted PAHs was measured by using a method developed in-house based on Randall hot extraction, purification by Solid Phase Extraction based on Molecular Imprinted Polymers, and Gas Chromatography Mass Spectrometry determination. A number of experimental studies were undertaken to generate data and information to improve the knowledge on migration of the target PAHs. Migration parameters operated in the STANPAHs project to estimate migration rates were as follows: dynamic mode at 40°C for 24 hours using a variety of migration media including artificial aqueous simulants, modified biosimulants formulations with lipidic content such as skin surface film liquid (SSFL), and 20% ethanol in water. According to scientific literature the use of 20% ethanol as the migration medium proved to correlate well with human skin absorption. Using these conditions, migration of the target PAHs into artificial sweat (EN1811) and artificial saliva (DIN53160-1) was not detected in any of the materials studied. Moreover none of the plastic polymeric materials led to detectable release of the target PAHs in any of the migration media used in this study (i.e. artificial sweat and saliva, skin surface film liquid (SSFL), and 20% ethanol solution). Similarly the tests with silicone materials did not result in detectable migration. Only the rubber matrices containing Distillate Aromatic Extract (DAE) as extender oil showed detectable migration when using 20% ethanol as the migration solution. In addition, the release of PAHs from coated recycled rubber granules was lower than from the uncoated granules suggesting that the coating acts as a barrier to chemical migration. According to industrial partners DAE is not used by European industries for manufacturing of parts of articles intended for skin contact. The materials containing DAE, although not representative for marketed products, have been made available to facilitate migration testing method development. The migration test method using 20% ethanol has been validated in-house and shows good method performance allowing the determination of PAH at trace level. Furthermore it has been considered for an initial inter-laboratory comparison study (ILC) aiming to investigate method applicability and transferability in a variety of laboratories. The within-laboratory precision, expressed as the relative standard deviation for repeatability (RSDr), and the between-laboratory precision, expressed as the relative standard deviation for reproducibility (RSDR) were assessed. In general the RSDR ranged from 28 to 113% and the RSDr from 7 to 23%. It is worth remembering that the level of PAH migration was very close to the quantification limit of the method and therefore this variability can be expected. Similar values have been reported in a recent German study with the participation of 9 laboratories on the migration of PAHs from rubber materials in contact with aqueous ethanol. The fact that better values of RSDr and RSDR were obtained for chrysene and benzo(e)pyrene that had the highest concentrations in the final migration solutions and that the analysis of the control solution used in this exercise showed a good reproducibility (RSDR% <10%), shows the possibility to reduce the variability between laboratories with a revised operating procedure in terms of injection volume and/or elution volume. In conclusion this report makes available new data and scientific information on the migration behaviour of certain PAHs from selected plastic and rubber polymeric matrices, in support of the European Commission's legal obligation to review the PAHs restriction under REACH. Standard operating procedures for quantification of the content of each of the eight restricted PAHs as well as their migration into 20% ethanol have been developed. Moreover the information gathered in STANPAHs (e.g. literature search), the ad-hoc manufactured materials still available, as well as the JRC in-house analysis method for PAH content could be of great benefit to accelerate the work towards standardisation of PAH content analysis in consumer products that has been recently undertaken by the European Standardisation Committee following a request by DG GROW.JRC.F.2-Consumer Products Safet

Digitalización y gestión de la infraestructura de recarga del vehículo eléctrico para su integración eficiente en la red de distribución

[ES] La integración del vehículo eléctrico (EV) es una pieza clave en la transición energética. Sin embargo, una alta penetración del EV supone un incremento de la demanda eléctrica con posibles consecuencias negativas sobre las redes de distribución. Para evitarlo, es necesario digitalizar las infraestructuras de recarga para ofrecer mecanismos de carga controlada con el objetivo de minimizar el impacto en la red y gestionar de manera activa y eficiente los recursos. En este artículo se presenta un piloto real de recarga con mecanismos de monitorización y de control de carga, en el que se muestran las ventajas de tener una infraestructura monitorizada, digitalizada y gestionada, así como los efectos que esto tiene sobre la red.La elaboración de este artículo se llevó a cabo en el marco de la red de Centros de Excelencia HySGrid+, con el título Desarrollo de comunidades locales energéticamente positivas con sistemas híbridos de generación renovable y almacenamiento financiado por el Ministerio de Ciencia, Innovación y Universidades a través del organismo CDTI, en el programa CER-20191019. Los autores agradecen la contribución del Instituto Valenciano de Competitividad Empresarial y del Fondo de Desarrollo Regional Europeo por hacer posible los desarrollos, permitiendo a ITE la difusión de los resultados y facilitando y promoviendo la transferencia de conocimiento a las empresas.Calatayud Martí, P.; Montagud Aguar, M.; Arcos Usero, L.; Romero-Chavarro, JC.; García Pellicer, M. (2020). Digitalización y gestión de la infraestructura de recarga del vehículo eléctrico para su integración eficiente en la red de distribución. Grupo Tecma Red. 138-143. http://hdl.handle.net/10251/178592S13814

Public Health–Driven Research and Innovation for Next-Generation Influenza Vaccines, European Union

Influenza virus infections are a major public health threat. Vaccination is available, but unpredictable antigenic changes in circulating strains require annual modification of seasonal influenza vaccines. Vaccine effectiveness has proven limited, particularly in certain groups, such as the elderly. Moreover, preparedness for upcoming pandemics is challenging because we can predict neither the strain that will cause the next pandemic nor the severity of the pandemic. The European Union fosters research and innovation to develop novel vaccines that evoke broadly protective and long-lasting immune responses against both seasonal and pandemic influenza, underpinned by a political commitment to global public health

Three-level methodology for secure and efficient grid integration of electric vehicle

[EN] The secure integration of electric vehicle (EV) plays a key role in the energy transition through a resilient and decarbonised economy. However, a massive EV penetration means a rise in electricity demand with negative consequences to the distribution systems (voltage drops, branches congestion, etc) if the charging infrastructure is not cybersecure and does not perform smart charging mechanisms. Furthermore, these new infrastructures and their operating procedures provide new chances to cyberattacks to be performed, aimed at either exploiting those grid vulnerabilities or acquiring some user's private information. Therefore, to ease the secure integration of EV charging infrastructures in the future network, this paper presents a three-level actuation methodology for charging infrastructures, which includes active management of EV supply equipment (EVSE) to allow dynamic control of charges, installation of ancillary protection systems, planning of EVSE's location within the distribution system and cybersecure management of the whole infrastructure. The presented methodology is based on a thorough analysis of the possible cyberattacks that may occur during the transactions of the charging process, as well as tests carried out on a real pilot, which demonstrate the possible impacts that an uncontrolled charging of the EV can have on the distribution network, thus identifying the vulnerabilities of the distribution network.The preparation of this article has been carried out within the framework of HySGrid+, a Spanish network of Centers of Excellence under the title "Development of energy-positive local communities with hybrid renewable generation and storage systems". HySGrid+ is funded by the Ministry of Science, Innovation and Universities through the CDTI agency, under the Cervera program "CER20191019". The authors of this article also thank the contribution of the "Instituto Valenciano de Competitividad Empresarial (IVACE)" and the European Regional Development Fund (ERDF) for making possible the development of this project, allowing the "Instituto Tecnológico de la Energía (ITE)" to disseminate the results of the research carried out and facilitating and promoting the transfer of knowledge to companies in the Valencian Community.Calatayud-Martí, P.; Romero-Chavarro, JC.; Montagud Aguar, M.; Arcos-Usero, L.; García Pellicer, M.; Quijano-Lopez, A. (2021). Three-level methodology for secure and efficient grid integration of electric vehicle. Dyna: Ingeniería e Industria. 96(3):264-269. https://doi.org/10.6036/1001326426996

Nanotechnology and the Environment

Nanotechnology has enormous potential for social, economic benefits, from innovative medical techniques to savings on materials and energy, as well as advances in detection and remediation of pollution. However, with environmental impacts as yet largely unknown and public controls absent, more systematic research and sector specific policies are necessary. Most of the current public dialogue on nanotechnology concerns its potential application and impacts on human health. From a socio-economic perspective, it is anticipated that the growth of nanotechnology may have positive or negative effects on specific communities and countries that are commodity dependent. For example, reduced resource extraction may benefit the environment but cause economic downturn. This is not unique to nanotechnology, but is typical of all technological innovation. Recognizing that these areas are inextricably linked, this paper focuses primarily on relatively uncharted territory: the relationship between nanotechnology and the environment. Many new technologies pose challenges for the environment. Today, scientific knowledge of environmental processes is significant, as is general awareness of environmental issues. Policy makers, industry, non-governmental organisations (NGOs) and scientists need to work together to raise public awareness of the opportunities and risks associated with nanotechnology. They also need to inform the public on whether steps are being undertaken to assess the potential impacts of nanomaterials before they enter the marketplace – as opposed to reacting after such materials become widespread. Nanotechnology creates many new possibilities for social and economic development, both in the short and long terms. The enhanced capacities to monitor the environment, to increase energy efficiency, and reduce the impact of human activities on the environment are clear potential benefits of the dissemination of nanomaterials. To realise this potential, more, not less, nanoscience and nanotechnology are needed. A balanced approach is required to maximise benefits while minimising risks. In this regard, a cooperative effort, which is both international and multi-sectoral, is strongly encouraged.JRC.H-Institute for environment and sustainability (Ispra

Impacto de la COVID-19 en el tratamiento del infarto agudo de miocardio con elevación del segmento ST. La experiencia Española

The COVID-19 outbreak has had an unclear impact on the treatment and outcomes of patients with ST-segment elevation myocardial infarction (STEMI). The aim of this study was to assess changes in STEMI management during the COVID-19 outbreak. Using a multicenter, nationwide, retrospective, observational registry of consecutive patients who were managed in 75 specific STEMI care centers in Spain, we compared patient and procedural characteristics and in-hospital outcomes in 2 different cohorts with 30-day follow-up according to whether the patients had been treated before or after COVID-19. Suspected STEMI patients treated in STEMI networks decreased by 27.6% and patients with confirmed STEMI fell from 1305 to 1009 (22.7%). There were no differences in reperfusion strategy (> 94% treated with primary percutaneous coronary intervention in both cohorts). Patients treated with primary percutaneous coronary intervention during the COVID-19 outbreak had a longer ischemic time (233 [150-375] vs 200 [140-332] minutes, P < .001) but showed no differences in the time from first medical contact to reperfusion. In-hospital mortality was higher during COVID-19 (7.5% vs 5.1%; unadjusted OR, 1.50; 95%CI, 1.07-2.11; P < .001); this association remained after adjustment for confounders (risk-adjusted OR, 1.88; 95%CI, 1.12-3.14; P = .017). In the 2020 cohort, there was a 6.3% incidence of confirmed SARS-CoV-2 infection during hospitalization. The number of STEMI patients treated during the current COVID-19 outbreak fell vs the previous year and there was an increase in the median time from symptom onset to reperfusion and a significant 2-fold increase in the rate of in-hospital mortality. No changes in reperfusion strategy were detected, with primary percutaneous coronary intervention performed for the vast majority of patients. The co-existence of STEMI and SARS-CoV-2 infection was relatively infrequent.S

Stress neuropeptide levels in adults with chest pain due to coronary artery disease: potential implications for clinical assessment

: Substance P (SP) and neuropeptide Y (NPY) are neuropeptides involved in nociception. The study of biochemical markers of pain in communicating critically ill coronary patients may provide insight for pain assessment and management in critical care. Purpose of the study was to to explore potential associations between plasma neuropeptide levels and reported pain intensity in coronary critical care adults, in order to test the reliability of SP measurements for objective pain assessment in critical care