European Regional Development Fund Research Articles

Consumption of ultra-processed foods and relationship with sodium and potassium excretion: a cross-sectional study

Introduction: Higher consumption of ultra-processed foods (UPF) had been associated with higher risk of non-communicable diseases (NCD) and NCD-promoting nutrient profile such as high sodium content and low potassium content. The objective of the study was to evaluate the level of food processing according to the NOVA classification and its relationship with sodium and potassium intake estimated by 24-hour urinary excretion. Methods: This is a cross-sectional study. Data collection included 107 workers from a public university (51.4% female; mean age 47 years) participating in an iMC Salt clinical trial). A 24-hour urinary collection was used to estimate sodium and potassium intake, validated by the creatinine coefficient. The corresponding 24-hour dietary recall was used to assess food intake and foods were categorized using the NOVA classification according to the degree and purpose of processing, into four groups. Nutri-score was also used to categorize UPF and provide more information about the nutritional quality of products. Results: The largest energy contribution came from the group of unprocessed or minimally processed foods (51.6%), followed by UPF (24%), processed foods (21.8%) and processed culinary ingredients (2.6%). Individuals with the lowest caloric contribution of the consumption of UPF had higher average values of potassium (K) intake (1438 vs 1136 mg/1000kcal; p=0.007), as individuals with the highest weight contribution of the consumption of unprocessed or minimally processed foods (1434 vs 1109 mg/1000kcal; p=0.010). For sodium (Na) intake no significant results were found between individuals with the lowest and the highest consumption of each NOVA group. Conclusions: Higher consumption of fresh or minimally processed foods and lower UPF intake were associated with higher K intake values. On the other hand, higher UPF intake was not associated with higher Na intake values. Funding: This work was supported by Fundação para a Ciência e Tecnologia by Grant POCI-01-0145-FEDER-029269) and by the European Regional Development Fund (ERDF) through NORTE 2020 (Northern Regional Operational Program 2014/2020) - NORTE-01-0145-FEDER-072687.

The Macroeconomic Effects of EU Regional Structural Funds

Abstract I examine the macroeconomic effects of two EU regional structural funds. The European regional development fund (ERDF) has positive short term average consequences, but gains typically dissipate within 3 years. The European Social Fund has insignificant average short term effects, but the medium term effect is positive and economically important. There are regional heterogeneities in terms of location, level of development, EU tenure, Euro area membership, and national borders. The reaction of regional government expenditure to shocks in the structural funds accounts for the differences. The implications for the EU Recovery and Resilience Facility are discussed.

Longitudinal profiling of human androgenotes through single-cell analysis unveils paternal gene expression dynamics in early embryo development.

How do transcriptomics vary in haploid human androgenote embryos at single cell level in the first four cell cycles of embryo development? Gene expression peaks at the fourth cell cycle, however some androcytes exhibit unique transcriptional behaviors. The developmental potential of an embryo is determined by the competence of the oocyte and the sperm. However, studies of the contribution of the paternal genome using pure haploid androgenotes are very scarce. This study was performed analyzing the single-cell transcriptomic sequencing of 38 androcytes obtained from 10 androgenote bioconstructs previously produced in vitro (de Castro et al., 2023). These results were analyzed through different bioinformatics software such as g: Profiler, GSEA, Cytoscape, and Reactome. Single cell sequencing was used to obtain the transcriptomic profiles of the different androcytes. The results obtained were compared between the different cycles studied using the DESeq2 program and functional enrichment pathways using g: Profiler, Cytoscape, and Reactome. A wave of paternally driven transcriptomic activation was found during the third-cell cycle, with 1128 upregulated and 225 downregulated genes and the fourth-cell cycle, with 1373 upregulated and 286 downregulated genes, compared to first-cell cycle androcytes. Differentially expressed routes related to cell differentiation, DNA-binding transcription, RNA biosynthesis and RNA polymerase II transcription regulatory complex, and cell death were found in the third and fourth with respect to the first-cell cycle. Conversely, in the fourth cell cycle, 153 downregulated and 332 upregulated genes were found compared with third cell cycle, associated with differentially expressed processes related to E-box binding and zinc finger protein 652 (ZNF652) transcription factor. Further, significant overexpression of LEUTX, PRAMEF1, DUXA, RFPL4A, TRIM43, and ZNF675 found in androgenotes, compared to biparental embryos, highlights the paternal contributions to zygote genome activation. All raw sequencing data are available through the Gene Expression Omnibus (GEO) under accessions number: GSE216501. Extrapolation of biological events from uniparental constructs to biparental embryos should be done with caution. Maternal and paternal genomes do not act independently of each other in a natural condition. The absence of one genome may affect gene transcription of the other. In this sense, the haploid condition of the bioconstructs could mask the transcriptomic patterns of the single cells. The results obtained demonstrated the level of involvement of the human paternal haploid genome in the early stages of embryo development as well as its evolution at the transcriptomic level, laying the groundwork for the use of these bioconstructs as reliable models to dispel doubts about the genetic role played by the paternal genome in the early cycles of embryo development. This study was funded by Instituto de Salud Carlos III (ISCIII) through the project 'PI22/00924', co-funded by European Regional Development Fund (ERDF); 'A way to make Europe'. F.D. was supported by the Spanish Ministry of Economy and Competitiveness through the Miguel Servet program (CPII018/00002). M.J.E. was supported by Instituto de Salud Carlos III (PI19/00577 [M.J.E.]) and FI20/00086. P.dC. was supported by a predoctoral grant for training in research into health (PFIS PI19/00577) from the Instituto de Salud Carlos III. All authors declare having no conflict of interest with regard to this trial.

Mechanical excavation of wetland habitat failed to eradicate invasive American red swamp crayfish (Procambarus clarkii) in Malta

Abstract Invasive crayfish are an important ecological concern in many freshwater ecosystems. Many efforts have been made to eradicate them, but there is very little documentation of the effectiveness of these efforts. Between 2019 and 2020, a restoration project funded by the European Regional Development Fund tried to eradicate invasive American red swamp crayfish Procambarus clarkii from the Fiddien valley system in Malta by mechanically excavating the valley's stream bed and exporting contaminated debris securely off‐site to a dry quarry. Three years post‐intervention, we systematically surveyed the valley system to explore the distribution and relative abundance of the invasive crayfish population. We placed traps in a stratified random sample of stream segments (both those that were included in the original restoration project and those that were not) and recorded catch per unit effort (crayfish caught per trap night) and the size/frequency distribution of crayfish caught. The invasive crayfish were still abundant in the upper reaches of the valley system, and, despite the excavation effort, crayfish were present at the highest relative abundance (4.3–14.8 CPUE, median = 12.3) within the restored area. Despite substantial effort and spending of more than 700,000 €, mechanical excavation did not eradicate invasive crayfish populations. We urge caution for future projects planning to attempt crayfish eradication using this approach and call for much greater impact evaluation, and at the very least post‐project monitoring, to ensure lessons can be learnt from such failures in future.

Specialization in climate technologies at the regional level in Europe: relatedness and the moderating role of policy direction

This study focuses on the influence of related and unrelated variety on specialization in climate change mitigation technologies on a regional level in Europe. We consider two dimensions of specialization: the degree of specialization and being specialized or not. We also investigate the moderating role of public policy direction towards climate change mitigation at regional, national, and EU regime level. Based on a fixed effect specification of patent data relating to climate technologies in 194 regions during the period 2008–2017, we find that related variety positively influences the degree of specialization in climate technologies, but only in regions with a low degree of specialization. We also report a weakening effect of national policy level environmental taxation and of the European Regional Development Fund for related variety. Regarding being specialized as a region in climate technologies, our study reveals a positive role played by unrelated variety and a negative moderating effect exerted by both national policy level environmental taxation and European Regional Development Fund policy support. We discuss implications for policymaking in the field of climate change mitigation technologies.

Selected aspects of perceptions of critical thinking among Polish and Czech primary school principals

The article concerns Polish-Czech research carried out as part of the project co-financed by the European Regional Development Fund and the state budget, We are crossing borders. 'Key competences in teaching and education' project was implemented under the Praděd Microprojects Fund. The descriptive purpose of the research was to prepare a detailed description of the level of competence of the surveyed people and the relationships between the dependent variable and independent variables.The explanatory purpose of the research was to find out why certain independent variables differentiate the respondents' answers.The study used the diagnostic survey method, the survey technique. They were attended by, among others, 49 principals from primary schools from Poland and the Czech Republic. The data analysis was carried out using the statistical program STATISTICA, in which statistical methods related to the study of the statistical significance of differences between variables were used using the Student's T-test for independent samples, to compare the means of two independent samples. Also the chi-square test was used.The results show how the management staff of educational institutions from Poland and the Czech Republic confidently perceive potential solutions to various difficult situations in their institutions and what critical thinking requirements they have for future teachers.

Climate change-related distributional range shifts of venomous snakes: a predictive modelling study of effects on public health and biodiversity

Climate change is expected to have profound effects on the distribution of venomous snake species, including reductions in biodiversity and changes in patterns of envenomation of humans and domestic animals. We estimated the effect of future climate change on the distribution of venomous snake species and potential knock-on effects on biodiversity and public health. We built species distribution models based on the geographical distribution of 209 medically relevant venomous snake species (WHO categories 1 and 2) and present climatic variables, and used these models to project the potential distribution of species in 2070. We incorporated different future climatic scenarios into the model, which we used to estimate the loss and gain of areas potentially suitable for each species. We also assessed which countries were likely to gain new species in the future as a result of species crossing national borders. We integrated the species distribution models with different socioeconomic scenarios to estimate which countries would become more vulnerable to snakebites in 2070. Our results suggest that substantial losses of potentially suitable areas for the survival of most venomous snake species will occur by 2070. However, some species of high risk to public health could gain climatically suitable areas for habitation. Countries such as Niger, Namibia, China, Nepal, and Myanmar could potentially gain several venomous snake species from neighbouring countries. Furthermore, the combination of an increase in climatically suitable areas and socioeconomic factors (including low-income and high rural populations) means that southeast Asia and Africa (and countries including Uganda, Kenya, Bangladesh, India, and Thailand in particular) could have increased vulnerability to snakebites in the future, with potential effects on public human and veterinary health. Loss of venomous snake biodiversity in low-income countries will affect ecosystem functioning and result in the loss of valuable genetic resources. Additionally, climate change will create new challenges to public health in several low-income countries, particularly in southeast Asia and Africa. The international community needs to increase its efforts to counter the effects of climate change in the coming decades. German Research Foundation, Conselho Nacional de Desenvolvimento Científico e Tecnológico, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, German Centre for Integrative Biodiversity Research, Ministerio de Ciencia e Innovación de España, European Regional Development Fund.

Lower respiratory tract single-cell RNA sequencing and neutrophil extracellular trap profiling of COVID-19-associated pulmonary aspergillosis: a single centre, retrospective, observational study

COVID-19-associated pulmonary aspergillosis (CAPA) is a severe superinfection with the fungus Aspergillus affecting patients who are critically ill with COVID-19. The pathophysiology and the role of neutrophil extracellular traps (NETs) in this infection are largely unknown. We aimed to characterise the immune profile, with a focus on neutrophils and NET concentrations, of critically ill patients with COVID-19, with or without CAPA. We conducted a single-centre, retrospective, observational study in two patient cohorts, both recruited at University Hospitals Leuven, Belgium. We included adults aged 18 years or older who were admitted to the intensive care unit because of COVID-19 between March 31, 2020, and May 18, 2021, and who were included in the previous Contagious trial (NCT04327570). We investigated the immune cellular landscape of CAPA versus COVID-19 only by performing single-cell RNA sequencing (scRNA-seq) on bronchoalveolar lavage fluid. Bronchoalveolar lavage immune cell fractions were compared between patients with CAPA and patients with COVID-19 only. Additionally, wedetermined lower respiratory tract NET concentrations using biochemical assays in patients aged 18 years and older who were admitted to the intensive care unit because of severe COVID-19 between March 15, 2020, and Dec31, 2021, for whom bronchoalveolar lavage was available in the hospital biobank. Bronchoalveolar lavage NET concentrations were compared between patients with CAPA and patients with COVID-19 only and integrated with existing data on immune mediators in bronchoalveolar lavage and 90-day mortality. We performed scRNA-seq of bronchoalveolar lavage on 43 samples from 39 patients, of whom 36 patients (30male and six female; 14 with CAPA) were included in downstream analyses. We performed bronchoalveolar lavage NET analyses in 59 patients (46 male and 13 female), of whom 26 had CAPA. By scRNA-seq, patients with CAPA had significantly lower neutrophil fractions than patients with COVID-19 only (16% vs 33%; p=0·0020). The remaining neutrophils in patients with CAPA preferentially followed a hybrid maturation trajectory characterised by expression of genes linked to antigen presentation, with enhanced transcription of antifungal effector pathways. Patients with CAPA also showed depletion of mucosal-associated invariant T cells, reduced T helper 1 and T helper 17 differentiation, and transcriptional defects in specific aspects of antifungal immunity in macrophages andmonocytes. We observed increased formation of NETs in patients with CAPA compared with patients with COVID-19 only (DNA complexed with citrullinated histone H3 median 15 898 ng/mL [IQR 4588-86 419] vs 7062 ng/mL [775-14 088]; p=0·042), thereby explaining decreased neutrophil fractions by scRNA-seq. Low bronchoalveolar lavage NET concentrations were associated with increased 90-day mortality in patients with CAPA. Qualitative and quantitative disturbances in monocyte, macrophage, B-cell, and T-cell populations could predispose patients with severe COVID-19 to develop CAPA. Hybrid neutrophils form a specialised response to CAPA, and an adequate neutrophil response to CAPA is a major determinant for survival in these patients. Therefore, measuring bronchoalveolar lavage NETs could have diagnostic and prognostic value in patients with CAPA. Clinicians should be wary of aspergillosis when using immunomodulatory therapy that might inhibit NETosis to treat patients with severe COVID-19. Research Foundation Flanders, KU Leuven, UZ Leuven, VIB, the Fundação para a Ciência e a Tecnologia, the European Regional Development Fund, la Caixa Foundation, the Flemish Government, and Horizon 2020.

Tick-borne diseases in the North Sea region–A comprehensive overview and recommendations for diagnostics and treatment

As part of the NorthTick project, co-funded by the European Union through the European Regional Development Fund and the North Sea Region Programme, specialists in the field of tick-borne diseases from seven North Sea countries co-operated with patient organisations and governmental health care institutions to provide this comprehensive overview of diagnostics and treatment recommendations in the region for Lyme borreliosis, Borrelia miyamotoi infection, tick-borne encephalitis, human granulocytic anaplasmosis, rickettsiosis, neoehrlichiosis and babesiosis. The main conclusion is that the recommendations in these northern countries are essentially the same, with very few differences. This overview presents the current diagnostics and provides useful clinical guidance.

The Impact of European Union Policies on Urban Development: A Case Study of Liverpool

This study delves into the history of urban development in Liverpool and the contemporary challenges it faces, examining the impact of European Union (EU) policies on its urban planning. It primarily focuses on employment opportunities and green policies, including transportation, clean energy, and biodiversity conservation. The research underscores the significant influence of the EU, mainly through the European Regional Development Fund (ERDF), in shaping Liverpool's development projects, enhancing spatial planning, and addressing urban issues. Recommendations for Liverpool City Council (LCC) include continued support for employment, especially among the diverse urban population, and exploring opportunities through development projects. The study emphasizes the need for LCC to align with EU policy directions, particularly in the context of the European Green Deal (EGD), to effectively address climate change sustainably. Post-Brexit, LCC should establish a committee to uphold and adapt EU policies to local needs, ultimately promoting Liverpool's resilience and sustainable urban development.

Systems biology analysis reveals distinct molecular signatures associated with immune responsiveness to the BNT162b COVID-19 vaccine

Human immune responses to COVID-19 vaccines display a large heterogeneity of induced immunity and the underlying immune mechanisms for this remain largely unknown. Using a systems biology approach, we longitudinally profiled a unique cohort of female high and low responders to the BNT162b vaccine, who were known from previous COVID-19 vaccinations to develop maximum and minimum immune responses to the vaccine. We utilized high dimensional flow cytometry, bulk and single cell mRNA sequencing and 48-plex serum cytokine analyses. We revealed early, transient immunological and molecular signatures that distinguished high from low responders and correlated with B and T cell responses measured 14 days later. High responders featured a distinct transcriptional activity of interferon-driven genes and genes connected to enhanced antigen presentation. This was accompanied by a robust cytokine response related to Th1 differentiation. Both transcriptome and serum cytokine signatures were confirmed in two independent confirmatory cohorts. Collectively, our data contribute to a better understanding of the immunogenicity of mRNA-based COVID-19 vaccines, which might lead to the optimization of vaccine designs for individuals with poor vaccine responses. German Center for Infection Research, German Center for Lung Research, German Research Foundation, Excellence Strategy EXC 2155 "RESIST" and European Regional Development Fund.

The Dynamics of Fund Absorption: Evaluating the Efficacy of EU Structural Funds in Mitigating Regional Inequalities—Calabrian Case

The European Union aims for territorial cohesion, with human capital as a key factor. Assessing how investment in regional human capital enhances this cohesion is therefore essential. This study assesses the impact of the EU Structural Funds (ESFs) in Calabria (Italy), a region grappling with economic challenges and a brain drain phenomenon. Aimed at fostering regional cohesion, ESFs have been directed towards supporting Calabrian graduates’ pursuit of master’s degrees, intending to incentivize their retention or return postgraduation. A comprehensive survey of the beneficiaries of these subsidies was carried out to determine their employability in the region and the probability of the return of migrants, analyzed by logistic regression of the data. Results demonstrate a dual effect: while the quality of education and EU funding positively influence graduates to work in Calabria, acquiring advanced skills paradoxically diminishes this propensity. Moreover, although the likelihood of returning to Calabria for those working elsewhere does increase, ESF support counterintuitively reduces this probability. The findings reveal a vicious cycle; they equip graduates with high-level skills that facilitate their access to the labor market but simultaneously encourage their migration due to more favorable conditions elsewhere. It is suggested that synergies between ESF-funded policies and those supported by the European Regional Development Fund (ERDF) should be encouraged.

Investigating the Use of Mg-3Pb Alloy for Cathodic Protection of Mg Alloys

Magnesium alloys possess many interesting properties, including high strength-to-weight ratios, good castability, and excellent biocompatibility. However, its use has been limited because they are also highly reactive and susceptible to corrosion in aqueous environments. As a result, the development of effective corrosion protection strategies for Mg alloys represents a significant challenge for many industrial applications, particularly in the aerospace, automotive, and biomedical industries.Sacrificial cathodic protection is a commonly used strategy to enhance the durability of metallic materials. This method involves the use of an active metal (known as the sacrificial anode) in galvanic contact with the metal to be protected from corrosion, which acts as the cathode. Due to their low corrosion potential, Mg alloys have traditionally served as sacrificial anodes to protect more noble metals such as steel, aluminum, and zinc. However, the growing interest in Mg alloys for industrial applications has led to the development of effective corrosion protection alternatives for Mg alloy components. Among them, the use of Mg alloy coatings as the anode material for the sacrificial protection of other Mg alloys is an interesting strategy. This approach offers several advantages such as minimized galvanic corrosion due to their comparable electrochemical behavior, as well as enhanced adhesion between the anode and the substrate.One of the key requirements for a metallic material to be used as a sacrificial anode is that its electrode potential must be more negative than that of the metal being protected. Additionally, the anode material must be chemically stable in the environment in which it will be located. These conditions are largely dependent on the composition of the anode material. Although recent studies have aimed to enhance the cathodic protection properties of Mg alloys for their use in industrial applications, such work remains restricted. This is due to the limited number of alloying elements with lower standard reduction potentials than Mg that are stable in solution, thereby decreasing the anode corrosion potential. In addition, Mg exhibits enhanced hydrogen evolution rates with increasing anodic polarization, which contradicts expectations of standard electrochemical kinetics and leads to significantly increased rates of self-dissolution. This phenomenon is termed anomalous hydrogen evolution (HE).This work investigates the potential of Mg-3Pb alloy as possible sacrificial anode material for the corrosion protection of three industrially important Mg alloys: AZ31, AM60, and AZ91. For that purpose, a Mg-3Pb alloy was casted in-house and its microstructural characteristics and electrochemical behavior were studied. Furthermore, galvanic corrosion between the Mg-3Pb alloy and the industrially relevant Mg alloys, AZ31, AM60, and AZ91, was evaluated in 0.1 M NaCl solution. The choice of Pb as an alloying element was motivated by its extremely low exchange current density for the HE reaction (i0,H2,Pb) on the order of 10-12 A/cm2. This was expected to result in reduced HE rates under cathodic polarization, which shifts the corrosion potential (Ecorr) to more negative values, and during anodic polarization, hindering anomalous HE and reducing self-dissolution.The microstructural characterization of the Mg-3Pb alloy revealed that it consisted of large α-Mg grains with the presence of a small amount of secondary phases, including Al, Si, Mn, Fe-containing intermetallics and oxide particles. Additionally, scanning electron microscopy revealed the occurrence of remarkable Pb segregation at grain boundaries.Electrochemical measurements showed that the Ecorr associated with the Mg-3Pb alloy was consistently lower than those of the AZ31, AM60, and AZ91 alloys during 24 hours of immersion in the test solution. Moreover, potentiodynamic polarization confirmed that the Mg-3Pb alloy exhibited the lowest cathodic kinetics. In terms of anomalous HE, which is directly related to the self-dissolution rate of the Mg alloy anode material, galvanostatic polarization at different anodic current densities showed that the HE current densities were linear with respect to the applied current density. Additionally, the charge associated with the HE was in the range of 40–50% of the applied anodic charge, which is consistent with previous findings for high purity Mg.Finally, ZRA measurements indicated that the galvanic current of the Mg-3Pb alloy remained anodic for 24 hours of immersion with no polarity reversal, indicating that the Pb-containing alloy served as sacrificial anode for the AZ31, AM60, and AZ91 alloys.In conclusion, the use of Mg-3Pb alloy in cathodic protection systems for Mg alloys has proved to be a viable option in 0.1 M NaCl solution, offering new possibilities for its use in various applications.Funding: This work was supported by the State Research Agency (MICINN), the Spanish National Research Council (CSIC) and European Regional Development Fund (ERDF) under the project RYC2019-027006-I (AEI/FEDER/UE).

Efficient High Temperature PEMFC Metallic Stack

High temperature (HT) polymer electrolyte membrane fuel cells (PEMFCs) are highly beneficial for high power applications [1-3]. They can be used both in stationary, as well as portable power devices [1,4,5] combining the advantages of the PEM type cells (flexibility in dynamic operation, compactness) and high temperature operation (elimination of fuel humidification and carbon monoxide clean up).This work focuses on the development, construction and testing of an efficient all-metallic HT-PEMFC stack with a power output of 1 kWe [6]. The selection of the proper polymer electrolytes allowed the operation of the stack within the range of 160-190oC. The Membrane-Electrode-Assemblies (MEA) and all components of the stack were designed and prepared in-house. The fuel cell stack characteristics and performance are affected by many parameters such as the materials, components’ design and proper assembly of the stack, as well as the operating conditions.The stack accomplished 50% electrical efficiency with power output of 1070 W and a power density of about 0.18 W/cm2 at 180°C under pure H2 feed. The same performance was reached with reformate hydrogen feeds. The 1 kW electrical and 1 kW thermal power HT-PEMFC stack presented herein consists of thirty MEAs, metallic bipolar plates and an innovative cost-efficient internal pumpless two-phase equilibrium water-steam cooling. The effective design and construction of the latter secures temperature gradientless and isothermal operation of the stack and the production of high temperature superheated steam, which is an effective carrier of high-quality thermal energy with several commercial and industrial applications. Acknowledgments Funding from Region of Western Greece and European Regional Development Fund (RIS3) through DemoCHP DЕР7-0019758 and the Operational Program “Competitiveness, Entrepreneurship & Innovation” (EPAnEK), NSRF 2014-2020 through Solar2HyP Τ2ΕΔΚ-01877.

Surface Curvature Effect on Dual-Atom Site Electrocatalysis

Oxygen, nitrogen, and carbon electrocatalysis are central in power-to-x innovations. Overpotentials of underlying processes are defined by the scaling relations between the adsorption energies of intermediates and, in turn, determine their efficiency. Our research explores the possibility of switching and tuning the scaling relations by applying metal-nitrogen-carbon (M-N-C) catalysts.In this presentation, we introduce dual-atom site models and demonstrate the effect of surface curvature on oxygen and nitrogen electrocatalysis. Our Density Functional Theory (DFT) calculations show how curving the surface switches the scaling and significantly lowers overpotentials. For instance, we predict a bifunctional overpotential as low as 0.45 V, markedly lower than the theoretical limit of 0.74 V given by the OOH–OH scaling. [1]Our study is among the first to investigate the surface curvature effect beyond the common scaling limitations. It will inspire the community to explore further this effect on oxygen electrocatalysis at M-N-C catalysts, as well as other challenging reactions and similarly curved materials. Funding V.I. and J.R. acknowledge the Danish National Research Foundation Centers of Excellence, The Center for High Entropy Alloys Catalysis (Project DNRF149), and the Independent Research Fund Denmark, grant no. 0217-00014B. V.I. receives funding from the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska–Curie grant agreement no. 101031656. This research was also supported by the Estonian Research Council grant PSG250 and the EU through the European Regional Development Fund (TK141).

Morphological and Electrical Characterization of AlGaN/GaN Heterostructures Modified By Atomic Layer Etching

Modern microelectronic components for high-power and high-frequency applications based on III-V compound semiconductors offer high break down voltage (GaN: 5 MV/cm and AlN: 15 MV/cm) at low specific on-resistance. However, it requires reliability and reproducibility of the production processes. Going to higher frequencies (5 GHz and beyond) and more integration in consumer electronics, smaller structure sizes are needed. This is going along with increasing demands on the manufacturing processes like dry etching. The main goal is to reach smaller and high controllable etching rates, going down to single atomic layers, low damage, and minimized surface roughness. Atomic layer etching (ALE) represents a key technology in order to achieve such requirements.An ALE process consists of two independent steps forming a repeatable cycle with a specific etching amount per cycle (EPC). First a surface modification is done by a chemical precursor i.e., adsorbing or oxidizing the surface. The second step is the removal of the modified layer, either physically by low energy non-reactive ion bombardment (Plasma Enhanced ALE [PEALE]) or chemically by a reactive species (Thermal ALE). The two steps are separated by an inert gas purge. This purging grants the removal of the chemistry after the first step, preventing further chemical reaction and transporting the chemical products out of the chamber after the second step.In this study, a PEALE conducted on different tools is used, for gate and contact recess study in a AlGaN/GaN heterostructure to form high electron mobility transistors (HEMT) with reduced ohmic contact resistance and threshold voltages. The results show a reduction of ohmic contact resistance at reduced forming temperatures and a linear dependence of the threshold voltage on gate recess depth. In order to determine the etching rate and roughness, atomic force microscopy (AFM) measurements were performed.The PEALE module is either integrated into a cluster tool consisting of atomic layer deposition (ALD) and chemical vapor deposition (CVD) or done in a combined ALD/ALE chamber. This opens the possibility of etching and depositing in-situ, reducing contamination and degradation of the substrate material.By this means, metal insulator semiconductor (MIS)-HEMTS with a recessed gate are studied. The monitoring of the individual processes is achieved by test structures like transfer length method structures (TLM) and metal oxide semiconductor (MOS), which showed the improved electric performance of these devices and may indicate unwanted interface issues, respectively. Additional, pre- and post-treatments could improve the interfaces further. The results will be presented and discussed in terms of ALE process development and optimization.This work was financially supported within the ALEStar project by the government of the Free-state of Saxony and the European Regional Development Fund under grant no. SAB 100402929.

Individualised, perioperative open-lung ventilation strategy during one-lung ventilation (iPROVE-OLV): a multicentre, randomised, controlled clinical trial

It is uncertain whether individualisation of the perioperative open-lung approach (OLA) to ventilation reduces postoperative pulmonary complications in patients undergoing lung resection. We compared a perioperative individualised OLA (iOLA) ventilation strategy with standard lung-protective ventilation in patients undergoing thoracic surgery with one-lung ventilation. This multicentre, randomised controlled trial enrolled patients scheduled for open or video-assisted thoracic surgery using one-lung ventilation in 25 participating hospitals in Spain, Italy, Turkey, Egypt, and Ecuador. Eligible adult patients (age ≥18 years) were randomly assigned to receive iOLA or standard lung-protective ventilation. Eligible patients (stratified by centre) were randomly assigned online by local principal investigators, with an allocation ratio of 1:1. Treatment with iOLA included an alveolar recruitment manoeuvre to 40 cm H2O of end-inspiratory pressure followed by individualised positive end-expiratory pressure (PEEP) titrated to best respiratory system compliance, and individualised postoperative respiratory support with high-flow oxygen therapy. Participants allocated to standard lung-protective ventilation received combined intraoperative 4 cm H2O of PEEP and postoperative conventional oxygen therapy. The primary outcome was a composite of severe postoperative pulmonary complications within the first 7 postoperative days, including atelectasis requiring bronchoscopy, severe respiratory failure, contralateral pneumothorax, early extubation failure (rescue with continuous positive airway pressure, non-invasive ventilation, invasive mechanical ventilation, or reintubation), acute respiratory distress syndrome, pulmonary infection, bronchopleural fistula, and pleural empyema. Due to trial setting, data obtained in the operating and postoperative rooms for routine monitoring were not blinded. At 24 h, data were acquired by an investigator blinded to group allocation. All analyses were performed on an intention-to-treat basis. This trial is registered with ClinicalTrials.gov, NCT03182062, and is complete. Between Sept 11, 2018, and June 14, 2022, we enrolled 1380 patients, of whom 1308 eligible patients (670 [434 male, 233 female, and three with missing data] assigned to iOLA and 638 [395 male, 237 female, and six with missing data] to standard lung-protective ventilation) were included in the final analysis. The proportion of patients with the composite outcome of severe postoperative pulmonary complications within the first 7 postoperative days was lower in the iOLA group compared with the standard lung-protective ventilation group (40 [6%] vs 97 [15%], relative risk 0·39 [95% CI 0·28 to 0·56]), with an absolute risk difference of -9·23 (95% CI -12·55 to -5·92). Recruitment manoeuvre-related adverse events were reported in five patients. Among patients subjected to lung resection under one-lung ventilation, iOLA was associated with a reduced risk of severe postoperative pulmonary complications when compared with conventional lung-protective ventilation. Instituto de Salud Carlos III and the European Regional Development Funds.

The involvement of GLP1 signaling with the effect of oral galactose in the hypothalamus of sporadic Alzheimer’s disease rat model

AbstractBackgroundIntracerebroventricular (icv) application of streptozotocin (STZ) causes mytochondrial dysfunction, oxidative stress and other metabolic effects, which leads to accumulation of τ‐protein and Aß plaques that makes it suitable for animal model of sporadic Alzheimer’s disease (sAD). We aimed to explore whether acute effect of single oral galactose dose is mediated by glucagon‐like peptide‐1 (GLP‐1) in sAD rat model.MethodWistar rats were injected icv with STZ (3 mg/kg) or vehicle only (CTR). One month after the injection, animals were divided into three experiments. First experiment: 200 mg/kg galactose was given orally and CTR animals were sacrificed 30, 60 and 120 min after administration. Second experiment: GLP‐1R antagonist (Exendin 9‐39, 85 µg/kg; GLP1Ra) was administered icv in both groups and animals were sacrificed 30 min after the injection. Third experiment: 30 min prior to oral galactose, rats were injected icv with GLP‐1Ra, and animals were sacrificed 120 min after galactose load. PDH, COXIV, CytC and Casp‐3 levels were measured by Western blot method and lactate, ATP and SOD were measured by ELISA in HPT. Glucose, galactose and GLP‐1 levels were measured in plasma and CSF. Data were analyzed by principal component analysis and Spearman rank correlation.ResultMetabolic changes were found more pronounced 120 min after single galactose load: reduced levels of Casp‐3 and PDH and increased SOD levels in HPT; increased CSF glucose. GLP1Ra decreased the levels of lactate, COXIV and CytC and increased PDH and Casp‐3 levels in HPT, only in STZ treated animals. CytC was found negatively correlated with ATP and PDH levels in HPT. GLP1Ra decreased the levels of CytC and SOD and increased PDH levels in CTR only after oral galactose.ConclusionGLP1Ra might inhibit decreased PDH levels caused by single galactose load. GLP1R central inhibition has more pronounced effect in STZ‐icv treated rats suggesting the involvement of GLP1 signaling in model pathogenesis. Supported by Croatian Science Foundation and co‐financed by the Scientific Centre of Excellence for Basic, Clinical and Translational Neuroscience (project “Experimental and clinical research of hypoxic‐ischemic damage in perinatal and adult brain”; GA KK01.1.1.01.0007 funded by the European Union through the European Regional Development Fund).

Label-Free Identification of Biochemical Footprints of Key Mutations in Acute Myeloid Leukemia Using Raman Spectroscopy

Introduction: With increasing understanding of the complexity and heterogeneity of acute myeloid leukemia (AML), the diagnostic process becomes more complicated and time-consuming. Therefore, there is a need to supplement AML diagnostics with a quick method to identify key, druggable genetic lesions that serve as a target for small molecule drugs. To address this need, we used Raman spectroscopy (RS)-based imaging methods. RS uses non-elastic scattering of laser light on cell biomolecules. When coupled to computational image segmentation and analysis, composition of the analyzed fragment of the cytoplasm can be characterized. In this way, by mapping the focal surface of the cell, we obtain a multidimensional image containing information about the distribution of its biochemical components. In our study, we utilized RS microscopy to investigate how single mutations affect the Raman spectral images of leukemic cells. The primary objective was to determine whether this technique could be utilized to identify mutations in key genes related to leukemia development. Methods: Genetic models of AML were generated by introducing wild-type and mutated forms of three frequently mutated and/or druggable genes (FLT3, IDH1/2, and MLL) into myeloid cell lines (THP1, HEL, HL60). The presence of the desired phenotype was confirmed using flow cytometry, immunoblotting, high-throughput kinase activity assay (Pamgene), and confocal microscopy. For RS experiments, the cells were fixed, and subjected to Raman imaging measurements using the WITec Alpha 300 microscope. Reference or control samples consisted of cells with wild-type genes (WT). The recorded hyperspectral data was subjected to multidimensional chemometric analysis, including k-means cluster analysis (KMCA), principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). To validate the findings from the cell models, primary AML cells from 30 patients diagnosed with FLT-3 ITD, MLL or IDH1/2 mutations were examined with RS. Peripheral blood mononuclear cells (PBMC) isolated from healthy donors blood served as reference. Results: The spectral differences between cells with wild-type FLT3 and FLT3/ITD were subtle, yet significant. The results revealed distinct variances in lipid (1440, 1240, 1660, and 2850 cm -1) and hemoprotein content (740-760 cm -1). Additionally, we successfully distinguished cells harboring IDH1/2 and MLL mutations from those with the wild-type genes. In these cases Raman bands corresponding to protein and nucleic acids composition were changed in comparison to control cells. Raman features in PCA loadings were also related to proteins and nucleic acids. Using the PLS-DA method and simple AI model, we built successful algorithm for distinguishing transgenic models carrying mutation from wild-type cells. PCA analysis also enabled us to distinguish AML primary cells from PBMC. Analysis weather primary cells carrying the selected mutations can be distinguished from non-mutated AML cells is concordant with our findings on models, however, it requires further analysis in larger group of patients to increase statistical power of these observations. Conclusions: Collectively, these findings suggest that Raman spectroscopy coupled with appropriate chemometric analysis may serve as rapid, label-free tool for identification of key druggable mutations in AML. Acknowledgements: The studies were performed as a part of the “ Label-free and rapid optical imaging, detection and sorting of leukemia cells” project carried out within the Team-Net programme of the Foundation for Polish Science co-financed by the European Union under the European Regional Development Fund.

Raman Spectroscopy Imaging in Evaluation of Immune Cells Activation Status

Introduction: The ability to assess the activation status of immune cells is useful for medical diagnostics and currently also for monitoring the performance of immune cells-based advanced therapy medicinal products (ATMP) such as the chimeric antigen receptor (CAR)-T cells. CAR-T cells numbers and immunophenotype are routinely monitored to avoid poor expansion or exhaustion phenotype. In this study, we utilized Raman spectroscopy (RS) as a label-free, non-destructive method for assessing T cells activation and macrophages polarization (pro-inflammatory M1 vs immunosuppressive M2). RS is a well-defined, non-elastic light scattering-based spectroscopic technique that followed by chemometric analysis provides unique and comprehensive information regarding the molecular structure, metabolic signature and biochemical composition of tissues and cells at subcellular level. To assess if RS would be a valid approach for detection of CAR-T activation we first developed CD19CAR-transduced Jurkat cell line model. Although Jurkat cells, a leukemic T cell line, lack cytolytic activity, they do produce IL-2 and upregulate CD69 upon activation like CAR-T thus serve as a convenient model. Materials and Methods: T cells and monocyte were isolated from healthy donors' blood. T cells were activated with coated CD3/CD28 beads. Jurkat cells were transduced with anti-CD19 chimeric TCR. CAR-J cells activation upon contact with CD19+ cells were confirmed by CD69 expression (flow cytometry) and calcium fluxes (fluorescence microscopy). Monocyte were differentiated to macrophages using CSF1, while THP1 monocytic cell line with PMA. Macrophages were subsequently polarized into M1 and M2 with IL-4 and IL-13 or IFN-gamma and LPS respectively. M0 macrophages or naïve T cells were used as a reference and control. Cell phenotypes were confirmed using flow cytometry, q-PCR, fluorescent confocal microscopy and proliferation assay. Cells were fixed in 0.5% glutaraldehyde and imaged at 533 and 633 nm with a WITec Alpha 300 Raman microscope. Collected data were analyzed by chemometric methods: K-means cluster analysis (KMCA) followed by Partial Least Squares-Discrimination Analysis (PLS-DA) and Principal Component Analysis (PCA). Results: RS analysis of polarized macrophages showed the increased signal from hemoproteins (740-760 cm -1) in all three states. Interestingly we observed different hemoproteins to lipids (1440, 1240, 1660, 2850 cm -1) ratio between M0, M1 and M2. Also degree of lipid unsaturation can be considered an important parameter in the identification of the M1 phenotype of macrophages. Chemometric analyzes revealed significant morphological (increased cell and nucleus sizes) biochemical and difference between activated T lymphocytes and naive cells. Carotenoids present in non-activated T cells disappeared after activation. Conversely, lipids in the form of distinct clusters were observed after activation. Moreover, in the spectra of cell nuclei (733, 790, 1583 cm -1) of activated lymphocytes, less condensed chromatin was manifested by a lower intensity of marker bands. We observed similar changes for activated CAR-Jurkat, though the morphological changes were more subtle and carotenoids were not observed in Jurkat cells. Conclusions: The obtained results indicate the potential utility of Raman spectroscopy methods in identifying lymphocyte states activation and macrophage polarization . Acknowledgements. The studies were performed as a part of the “ Label-free and rapid optical imaging, detection and sorting of leukemia cells” project carried out within the Team-Net programme of the Foundation for Polish Science co-financed by the European Union under the European Regional Development Fund.