Surface Plasmon Resonance kinetic analysis of the interaction between G-quadruplex nucleic acids and an anti-G-quadruplex monoclonal antibody

Background G-quadruplexes (G4s) are nucleic acids secondary structures formed in guanine-rich sequences. Anti-G4 antibodies represent a tool for the direct investigation of G4s in cells. Surface Plasmon Resonance (SPR) is a highly sensitive technology, suitable for assessing the affinity between biomolecules. We here aimed at improving the orientation of an anti-G4 antibody on the SPR sensor chip to optimize detection of binding antigens. Methods SPR was employed to characterize the anti-G4 antibody interaction with G4 and non-G4 oligonucleotides. Dextran-functionalized sensor chips were used both in covalent coupling and capturing procedures. Results The use of two leading molecule for orienting the antibody of interest allowed to improve its activity from completely non-functional to 65% active. The specificity of the anti-G4 antobody for G4 structures could thus be assessed with high sensitivity and reliability. Conclusions Optimization of the immobilization protocol for SPR biosensing, allowed us to determine the anti-G4 antibody affinity and specificity for G4 antigens with higher sensitivity with respect to other in vitro assays such as ELISA. Anti-G4 antibody specificity is a fundamental assumption for the future utilization of this kind of antibodies for monitoring G4s directly in cells. General significance The heterogeneous orientation of amine-coupling immobilized ligands is a general problem that often leads to partial or complete inactivation of the molecules. Here we describe a new strategy for improving ligand orientation: driving it from two sides. This principle can be virtually applied to every molecule that loses its activity or is poorly immobilized after standard coupling to the SPR chip surface

Visualization of DNA G-quadruplexes in herpes simplex virus 1-infected cells

We have previously shown that clusters of guanine quadruplex (G4) structures can form in the human herpes simplex-1 (HSV-1) genome. Here we used immunofluorescence and immune-electron microscopy with a G4-specific monoclonal antibody to visualize G4 structures in HSV-1 infected cells. We found that G4 formation and localization within the cells was virus cycle dependent: viral G4s peaked at the time of viral DNA replication in the cell nucleus, moved to the nuclear membrane at the time of virus nuclear egress and were later found in HSV-1 immature virions released from the cell nucleus. Colocalization of G4s with ICP8, a viral DNA processing protein, was observed in viral replication compartments. G4s were lost upon treatment with DNAse and inhibitors of HSV-1 DNA replication. The notable increase in G4s upon HSV-1 infection suggests a key role of these structures in the HSV-1 biology and indicates new targets to control both the lytic and latent infection

Exploring drivers of consumer well-being in music festivals: a relationship marketing approach

In the recent decades, the music festivals industry has developed very fast as well as contributed for the growth of wealth in the tourism sector. However, the research up-to-date about festivals is at an early stage, as there are plenty concepts and themes that have not been explored yet. Therefore, this study is motivated to explore what is behind the music festivals’ phenomenon. This dissertation considers the context of music festivals in Portugal due to its increasing levels of tourism growth, and investigates the existing relationships between common concepts within the field of Relationship Marketing, such as consumer’s experience, emotions and level of satisfaction, and yet their level engagement and their sense of well-being. In order to establish a connection between all these constructs, the S-O-R framework developed by Mehrabian and Russell (1974), regarded as one of the most influential in environmental psychology, was adapted to this study to carry this investigation. The results of the study revealed that all the constructs explored in the study show a positive relationship that enables to better comprehend the music festival experience felt by visitors, but it should be noted that not all the dimensions of each construct can be considered as relevant to our theoretical model. The overall findings in the Conclusion Chapter presented some interesting information, particularly, the relationship between consumers’ engagement and well-being, because it provides innovative and revealing facts that have not been explored previously in the music festivals literature. The managerial implications concerned to this study are discussed at the end.Nas últimas décadas, a indústria dos festivais de música tem vindo a desenvolver-se rapidamente, como ainda tem contribuído para o crescimento da riqueza que é gerada através do sector do turismo. No entanto, a pesquisa que é recolhida até à data acerca de festivais encontra-se ainda numa fase inicial, uma vez que existem vários conceitos e temas que não foram ainda investigados. Desta forma, este estudo é motivado a explorar o que está por detrás do fenómeno dos festivais de música. Esta dissertação considera o contexto dos festivais de música em Portugal, devido aos altos níveis de crescimento do seu turismo, e investiga as relações entre alguns conceitos que são comuns na área do Marketing Relacional, como por exemplo, a experiência dos consumidores, as suas emoções e satisfação, e ainda os seus níveis de compromisso, e as suas sensações de bem-estar. De maneira a estabelecer uma conexão entre todos estes conceitos, o framework S-O-R desenvolvido por Mehrabian e Russell (1974), reconhecido como um dos mais influentes na área de psicologia ambiental, foi adaptado a este estudo para que fosse conduzida esta investigação. Os resultados deste estudo revelaram que todos os constructos explorados neste estudo demonstram uma relação positiva que permite compreender melhor a experiência dos festivais de música sentida pelos visitantes, mas deve também ser referido que nem todas as dimensões de cada constructo podem ser consideradas como relevantes para este modelo teórico. Os resultados gerais que se encontram no capítulo das conclusões apresentaram informação relevante, nomeadamente, a relação entre compromisso e bem-estar dos consumidores, por apresentar factos inovadores e reveladores que não foram ainda explorados na literatura dos festivais de música. As implicações que foram reconhecidas neste estudo são discutidas no final do mesmo

Synthesis, Binding and Antiviral Properties of Potent Core-Extended Naphthalene Diimides Targeting the HIV-1 Long Terminal Repeat Promoter G-Quadruplexes

We have previously reported that stabilization of the G-quadruplex structures in the HIV-1 long terminal repeat (LTR) promoter suppresses viral transcription. Here we sought to develop new G-quadruplex ligands to be exploited as antiviral compounds by enhancing binding toward the viral G-quadruplex structures. We synthesized naphthalene diimide derivatives with a lateral expansion of the aromatic core. The new compounds were able to bind/stabilize the G-quadruplex to a high extent, and some of them displayed clear-cut selectivity toward the viral G-quadruplexes with respect to the human telomeric G-quadruplexes. This feature translated into low nanomolar anti-HIV-1 activity toward two viral strains and encouraging selectivity indexes. The selectivity depended on specific recognition of LTR loop residues; the mechanism of action was ascribed to inhibition of LTR promoter activity in cells. This is the first example of G-quadruplex ligands that show increased selectivity toward the viral G-quadruplexes and display remarkable antiviral activity

Factors That Impact the Relationship between Perceived Organizational Support and Technostress in Teachers

In the last two years, the obligatory use of technologies due to the COVID-19 pandemic has increased the technostress suffered by education professionals. This study investigates the relationships between technostress and perceived organizational support and the influence of certain socio-demographic variables. An online survey was administered to 771 teachers working in different educational stages in various autonomous communities in Spain. Perceived organizational support was found to be significantly correlated with technostress. Women tend to experience more technostress in general and significant gender differences were also found in the dimension of anxiety. The analyzed data also suggest that perceived organizational support is higher in private schools. In urban centers, teachers’ technostress increases in higher educational stages, such as secondary education and baccalaureate. Further work is needed to develop school policies that address the needs of teachers and provide support for those at risk of technostress. In addition, there is a need to design coping strategies and prioritize the most at-risk sectors to improve their overall health and well-being

Migrant workers and the digital transformation in the EU

The aim of this report is to provide insights on the implications that structural changes in the labour market related to the Digital Transformation (DT) could have on the integration of EU mobile citizens and third country nationals working in the EU. A comprehensive analysis of the changing nature of the EU labour markets and the effects of DT is provided in the upcoming European Commission’s 2018 Employment and Social Developments in Europe (ESDE) review. Building upon these general findings, this report contributes to the debate from a migration-specific point of view by providing evidence on the extent to which migrants are employed in occupations that are potentially prone to automation and therefore may disappear in future. The analysis is based on data drawn from EU LFS 2015-2016 and PIAAC 2012 surveys. The results show that: • Third country nationals tend to be more concentrated in occupations characterized by high routine intensity and thus more prone to automation (e.g. elementary occupations), followed by EU mobile citizens and by natives. • Both EU mobile citizens and third country nationals have a higher likelihood of being employed in jobs with high automation potential than nationals, even when socio-demographic characteristics are taken into account. However, the likelihood decreases as educational attainment increases, for all but more so for migrants. • Major differences between EU mobile citizens and third country nationals appear when considering their length of residence. The results show that among EU mobile citizens, recent migrants have higher odds of being employed in a job with high automation potential compared to long-term migrants. On the contrary, in the case of third country nationals, long term migrants report higher odds of working in a job with high automation potential than recent migrants. • Both EU mobile citizens and third country nationals are less likely to receive professional training in comparison to nationals. This lower investment in the human capital of migrants can hamper migrants’ opportunities to transition to other jobs once they would lose their jobs due to the DT. • Both EU mobile citizens and third country nationals are more likely to be on fixed-term contracts with a shorter horizon compared to natives, with risk of non-renewal of contract in case of economic and technological shocks. • In summary, the vulnerability of migrants in the labour market is furthermore reinforced by the fact that they tend to be concentrated in jobs with high automation potential which, in turn, are associated to lower training and more widespread use of fixed-term contracts.JRC.E.6-Demography, Migration and Governanc

Environmental and economic benefits of wheat and chickpea crop rotation in the Mediterranean region of Apulia (Italy)

Wheat plays an essential role in safeguarding global food security. However, its intensive agricultural production, aimed at maximizing crop yields and associated economic benefits, jeopardizes many ecosystem services and the economic stability of farmers. Rotations with leguminous are recognized as a promising strategy in favor of sustainable agriculture. However, not all crop rotations are suitable for promoting sustainability and their implications on agricultural soil and crop quality should be carefully analyzed. This research aims to demonstrate the environmental and economic benefits of introducing chickpea into a wheat-based system under Mediterranean pedo-climatic conditions. For this purpose, the crop rotation “wheat-chickpea” was evaluated and compared with the conventional regime (wheat monoculture) by means of life cycle assessment methodology. For this purpose, inventory data (e.g., agrochemical doses, machinery, energy consumption, production yield, among others) was compiled for each crop and cropping system, thus converted into environmental impacts based on two functional units: 1 ha per year and one € of gross margin. Eleven environmental indicators were analyzed, including soil quality and biodiversity loss. Results indicate that chickpea-wheat rotation system offers lower environmental impacts, regardless of the functional unit considered. Global warming (18 %) and freshwater ecotoxicity (20 %) were the categories with the largest reductions. Furthermore, a remarkable increase (96 %) in gross margin was observed with the rotation system, due to the low cost of chickpea cultivation and its higher market price. Nevertheless, proper fertilizer management remains essential to fully attain the environmental benefits of crop rotation with legumesThis research is supported by the project Enhancing diversity in Mediterranean cereal farming systems (CerealMed), funded by PRIMA Program and FEDER/Ministry of Science and Innovation – Spanish National Research Agency (PCI2020-111978) and the project Transition to sustainable agri-food sector bundling life cycle assessment and ecosystem services approaches (ALISE), funded by the Spanish National Research Agency (TED2021-130309B-I00). S.L.O., R.R.L, M.T.M. and S.G.G belong to the Galician Competitive Research Group (GRC ED431C-2021/37) and to the Cross-disciplinary Research in Environmental Technologies (CRETUS Research Center, ED431E 2018/01)S

Improving environmental sustainability of agriculture in Egypt through a life-cycle perspective

Soil plays an essential role as a habitat, source of nutrients and support for vegetation. Promoting food security and environmental sustainability of agricultural systems requires an integrated approach to soil fertility management. Agricultural activities should be developed with preventive approaches aimed at avoiding or reducing negative impacts on the soil physicochemical and biological properties and the depletion of soil nutrient reserves. In this regard, Egypt has developed the Sustainable Agricultural Development Strategy to encourage environmentally friendly practices among farmers, such as crop rotation and water management, in addition to extending agriculture to desert areas, favoring the socio-economic development of the region. In order to evaluate the outcomes of the plan beyond quantitative data of production, yield, consumption and emissions, the environmental profile of agriculture in Egypt has been assessed under a life-cycle perspective in order to identify the associated environmental burdens and ultimately contribute to improving the sustainability policies of agricultural activity within the framework of a crop rotation system. In particular, a two-year crop rotation (Egyptian clover-maize-wheat) was analyzed in two distinct agricultural areas in Egypt: New Lands in desert regions and Old Lands along the Nile River, traditionally recognized as fertile areas due to the river alluvium and water availability. The New Lands had the worst environmental profile for all impact categories, except for Soil organic carbon deficit and Global potential species loss. Irrigation and on-field emissions associated with mineral fertilization were identified as the most critical hotspots of Egyptian agriculture. In addition, land occupation and land transformation were reported as the main drivers of biodiversity loss and soil degradation, respectively. Beyond these results, further research on biodiversity and soil quality indicators is needed to more accurately assess the environmental damage caused by the conversion of deserts into agricultural areas, given the species richness these regions holdThis research is supported by the project Enhancing diversity in Mediterranean cereal farming systems (CerealMed), funded by PRIMA Programme, FEDER/Ministry of Science and Innovation– Spanish National Research Agency (PCI2020-111978); and Academy of Scientific Research and Technology (ASRT); and the project Transition to sustainable agri-food sector bundling life cycle assessment and ecosystem services approaches (ALISE), funded by the Spanish National Research Agency (TED2021-130309B-I00). S.L.O., M.T.M. and S.G.G belong to the Galician Competitive Research Group (GRC ED431C-2021/37) and to the Cross-disciplinary Research in Environmental Technologies (CRETUS Research Center, ED431E 2018/01)This research is supported by the project Enhancing diversity in Mediterranean cereal farming systems (CerealMed), funded by PRIMA Programme, FEDER/Ministry of Science and Innovation– Spanish National Research Agency (PCI2020-111978); and Academy of Scientific Research and Technology (ASRT); and the project Transition to sustainable agri-food sector bundling life cycle assessment and ecosystem services approaches (ALISE), funded by the Spanish National Research Agency (TED2021-130309B-I00). S.L.O., M.T.M. and S.G.G belong to the Galician Competitive Research Group (GRC ED431C-2021/37) and to the Cross-disciplinary Research in Environmental Technologies (CRETUS Research Center, ED431E 2018/01)S