Quantitative analysis of the benzanthrone aminoderivative binding to amyloid fibrils of lysozyme

The accumulation of amyloid fibrils in different tissues is associated with a number of neurodegenerative diseases. Despite a huge variety of amyloid-specific probes, all of them suffer from many drawbacks, highlighting the necessity of searching for more preferable dyes. In the present work, the potential of new fluorescent probe AM3 for selective detection of fibrillar protein aggregates, formed from lysozyme, has been evaluated. To quantify the affinity of this dye for amyloid fibrils, the isotherms of dye binding to the fibrillar lysozyme have been derived from fluorimetric titration. Parameters of the dye-protein complexation: association constant, molar fluorescence and binding stoichiometry, calculated from the Langmuir adsorption model, revealed that AM3 interacts strongly with protein insoluble aggregates. High values of the binding parameters make AM3 an alternative to a widely-used amyloid-specific probe Thioflavin T. We also investigated the effects of polarity and viscosity on AM3 fluorescence properties. The binding of AM3 to the protein hydrophobic cavities has been followed by red shift of the dye emission spectra, which can be explained by H-bonding between proton-donating groups of the protein and carbonyl moiety of the probe. Long-wavelength shift of emission maximum was observed also upon increasing the excitation wavelength. This finding suggests that reorientation time of solvent molecules is higher, than the dye fluorescence lifetime. Fluorescence anisotropy studies revealed slow rotation diffusion of the probe, bound to amyloid fibrils being indicative of high viscosity of AM3 microenvironment. The observed photophysical properties of the new aminobenzanthrone derivative make AM3 a perspective probe for basic research and medical diagnostics

Interaction of novel benzanthrone derivative with amyloid lysozyme

A novel benzanthrone derivative AM18 was investigated with respect to its photophysical properties when bound to native, oligomeric and fibrillar hen egg white lysozyme. As shown by fluorimetric titration AM18 is more sensitive to pathogenic protein aggregates than Thioflavin T, however has no ability to differentiate between mature and immature lysozyme fibrils. The recovered affinity and fluorescence response of the novel probe to amyloid protein appeared to be similar to those of recently developed amyloid lysozyme-sensitive dyes like e. g. Nile Red and cyanine dye 7515. Despite the high increase of the probe emission in the presence of amyloid lysozyme compared to its fluorescence in buffer, the minimal amount that could be detected by 1 μM AM18 was 10 times lower for amyloid-native protein solutions due to high affinity of the dye for lysozyme monomers. In general, because of high quantum yields and “signal-to-noise” ratios in the presence of pathogenic protein aggregates AM18 appeared to be an effective tool for amyloid detection and characterization in vitro, being however unable to detect pathogenic protein aggregates in vivo like e.g. recently reported p-FTAA because of the sensitivity to lipids. Compared to previously reported AM3 a novel dye showed 2-fold lower “signal-to- noise” ratio in the presence of fibrillar lysozyme, and 2 fold lower blue shift of emission maximum. This tendency was explained in terms of decreased charge transfer from the donor to acceptor groupes of AM18 compared to AM3. Finally, as concluded from the comparison of AM18 and previously studied benzanthrone derivatives, the 5 nm – red edge excitation shift of AM18 is indicative of its possible binding to fibril “deep cavities”, containing no water. High anisotropy values of amyloid-bound dye led us to conclusion that the enhanced fluorescence of the probe is associated with the decrease of the rotational motion of the amino-substitute about the benzanthrone unit. This is a sign of AM18 behaviour as a molecular rotor

Analysis of factors that influence eating habits in different countries

Individual eating habits are influenced by a number of factors, including both internal variables such as physiology and emotion, as well as environmental factors such as food availability and cultural norms. Given the public health impact of dietary habits (choice, quality, amount, frequency) on health outcomes, it is important to understand what factors influence eating habits on a societal level. The aim of this research was to determine factors that influence eating habits and compare these factors between four different countries – Latvia, Lithuania, Portugal and the USA. An eating motivation questionnaire was used to measure eating habits in 3,348 respondents from different regions and countries. There were ten parts - demographical information, anthropometric data and behavioral and health related elements, sources of information about healthy eating, factors related to food choices according to motivations (health, emotional, economic, availability, social, cultural, environmental, political, marketing and commercials). Data were analyzed using descriptive statistics, and self-reported motivation was compared across countries. Health was the primary motivator of food selection in this sample (71% of respondents), whereas 34% reported that emotional factors impact their dietary habits and 35% reported that economic factors determine their food selection. A large number of respondents (44%) disagreed or strongly disagree or disagreed with the idea that marketing impacts their dietary habits. Portugal had the highest number of participants (86%), reporting that they agreed or strongly agreed with having health-related motivations for food selection, with Latvia (65%) and Lithuania (76%) showing more moderate levels of endorsement of healthy eating motivations, and the USA having the fewest respondents (52%) endorsing health-related motivations. Respondents from Portugal were more likely than respondents from the other countries to deny having emotional, economic and marketing motivations in food selection. From results can conclude that consumers are motivated by healthiness factors when making food choices (71% of respondents), but marketing, economic and emotional factors positively impact only 30% of consumers, other respondents completely disagreed or was indifferent to these types of motivations. Baltic countries (Latvia and Lithuania) were similar to each other, but Portugal and USA were completely different. Portugal strongly agreed with healthy motivations and disagreed with all other motivations, whereas USA and also Baltic countries had more equal division of opinions regarding impact of different motivations.info:eu-repo/semantics/publishedVersio

Reverse engineering of model transformations for reusability

The final publication is available at Springer via http://dx.doi.org/10.1007/978-3-319-08789-4_14Proceedings of 7th International Conference, ICMT 2014, Held as Part of STAF 2014, York, UK, July 21-22, 2014Reuse techniques are key for the industrial adoption of Model-Driven Engineering (MDE). However, while reusability has been successfully applied to programming languages, its use is scarce in MDE and, in particular, in model transformations. In previous works, we developed an approach that enables the reuse of model transformations for different meta-models. This is achieved by defining reusable components that encapsulate a generic transformation template and expose an interface called concept declaring the structural requirements that any meta-model using the component should fulfil. Binding the concept to one of such meta-models induces an adaptation of the template, which becomes applicable to the meta-model. To facilitate reuse, concepts need to be concise, reflecting only the minimal set of requirements demanded by the transformation. In this paper, we automate the reverse engineering of existing transformations into reusable transformation components. To make a transformation reusable, we use the information obtained from its static analysis to derive a concept that is minimal with respect to the transformation and maximizes its reuse opportunities, and then evolve the transformation accordingly. The paper describes a prototype implementation and an evaluation using transformations from the ATL zoo.This work has been funded by the Spanish Ministry of Economy and Competitivity with project “Go Lite” (TIN2011-24139

Tritium in plasma-facing components of JET with the ITER-Like-Wall

Publisher Copyright: © 2021 Institute of Physics Publishing. All rights reserved.The ITER-Like-Wall project has been carried out at the Joint European Torus (JET) to test plasma facing materials relevant to ITER. Materials being tested include both bulk metals (Be andW) and coatings. Tritium accumulation mechanisms and release properties depend both on the wall components, their location in the vacuum vessel, conditions of exposure to plasma and to the material itself. In this study, bulk beryllium limiter tiles, plasma-facing beryllium coated Inconel components from the main chamber, bulk tungsten and tungsten coated carbon fibre composite divertor tiles were analysed. A range of methods have been developed and applied in order to obtain a comprehensive overview on tritium retention and behaviour in different materials of plasma facing components (PFCs). Tritium content and chemical state were studied by the means of chemical or electrochemical dissolution methods and thermal desorption spectroscopy. Tritium distribution in the vacuum vessel and factors affecting its accumulation have been assessed and discussed.publishersversionPeer reviewe

Pollen-based quantitative reconstructions of Holocene regional vegetation cover (plant-functional types and land-cover types) in Europe suitable for climate modelling

We present quantitative reconstructions of regional vegetation cover in north-western Europe, western Europe north of the Alps, and eastern Europe for five time windows in the Holocene [around 6k, 3k, 0.5k, 0.2k, and 0.05k calendar years before present (bp)] at a 1 degrees x1 degrees spatial scale with the objective of producing vegetation descriptions suitable for climate modelling. The REVEALS model was applied on 636 pollen records from lakes and bogs to reconstruct the past cover of 25 plant taxa grouped into 10 plant-functional types and three land-cover types [evergreen trees, summer-green (deciduous) trees, and open land]. The model corrects for some of the biases in pollen percentages by using pollen productivity estimates and fall speeds of pollen, and by applying simple but robust models of pollen dispersal and deposition. The emerging patterns of tree migration and deforestation between 6k bp and modern time in the REVEALS estimates agree with our general understanding of the vegetation history of Europe based on pollen percentages. However, the degree of anthropogenic deforestation (i.e. cover of cultivated and grazing land) at 3k, 0.5k, and 0.2k bp is significantly higher than deduced from pollen percentages. This is also the case at 6k in some parts of Europe, in particular Britain and Ireland. Furthermore, the relationship between summer-green and evergreen trees, and between individual tree taxa, differs significantly when expressed as pollen percentages or as REVEALS estimates of tree cover. For instance, when Pinus is dominant over Picea as pollen percentages, Picea is dominant over Pinus as REVEALS estimates. These differences play a major role in the reconstruction of European landscapes and for the study of land cover-climate interactions, biodiversity and human resources.Peer reviewe

Identifying Alternative Hyper-Splicing Signatures in MG-Thymoma by Exon Arrays

BACKGROUND: The vast majority of human genes (>70%) are alternatively spliced. Although alternative pre-mRNA processing is modified in multiple tumors, alternative hyper-splicing signatures specific to particular tumor types are still lacking. Here, we report the use of Affymetrix Human Exon Arrays to spot hyper-splicing events characteristic of myasthenia gravis (MG)-thymoma, thymic tumors which develop in patients with MG and discriminate them from colon cancer changes. METHODOLOGY/PRINCIPAL FINDINGS: We combined GO term to parent threshold-based and threshold-independent ad-hoc functional statistics with in-depth analysis of key modified transcripts to highlight various exon-specific changes. These denote alternative splicing in MG-thymoma tumors compared to healthy human thymus and to in-house and Affymetrix datasets from colon cancer and healthy tissues. By using both global and specific, term-to-parent Gene Ontology (GO) statistical comparisons, our functional integrative ad-hoc method allowed the detection of disease-relevant splicing events. CONCLUSIONS/SIGNIFICANCE: Hyper-spliced transcripts spanned several categories, including the tumorogenic ERBB4 tyrosine kinase receptor and the connective tissue growth factor CTGF, as well as the immune function-related histocompatibility gene HLA-DRB1 and interleukin (IL)19, two muscle-specific collagens and one myosin heavy chain gene; intriguingly, a putative new exon was discovered in the MG-involved acetylcholinesterase ACHE gene. Corresponding changes in spliceosome composition were indicated by co-decreases in the splicing factors ASF/SF(2) and SC35. Parallel tumor-associated changes occurred in colon cancer as well, but the majority of the apparent hyper-splicing events were particular to MG-thymoma and could be validated by Fluorescent In-Situ Hybridization (FISH), Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and mass spectrometry (MS) followed by peptide sequencing. Our findings demonstrate a particular alternative hyper-splicing signature for transcripts over-expressed in MG-thymoma, supporting the hypothesis that alternative hyper-splicing contributes to shaping the biological functions of these and other specialized tumors and opening new venues for the development of diagnosis and treatment approaches

Metformin strongly affects transcriptome of peripheral blood cells in healthy individuals

Funding Information: The study was supported by the European Regional Development Fund under the project ?Investigation of interplay between multiple determinants influencing response to metformin: search for reliable predictors for efficacy of type 2 diabetes therapy? (Project No.: 1.1.1.1/16/A/091, https://ec.europa.eu/regional_policy/en/funding/ erdf/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors would like to thank all the volunteers for their participation and acknowledge the Genome Database of the Latvian Population for providing biological material and data. Publisher Copyright: © 2019 Ustinova et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Metformin is a commonly used antihyperglycaemic agent for the treatment of type 2 diabetes mellitus. Nevertheless, the exact mechanisms of action, underlying the various therapeutic effects of metformin, remain elusive. The goal of this study was to evaluate the alterations in longitudinal whole-blood transcriptome profiles of healthy individuals after a one-week metformin intervention in order to identify the novel molecular targets and further prompt the discovery of predictive biomarkers of metformin response. Next generation sequencing-based transcriptome analysis revealed metformin-induced differential expression of genes involved in intestinal immune network for IgA production and cytokine-cytokine receptor interaction pathways. Significantly elevated faecal sIgA levels during administration of metformin, and its correlation with the expression of genes associated with immune response (CXCR4, HLA-DQA1, MAP3K14, TNFRSF21, CCL4, ACVR1B, PF4, EPOR, CXCL8) supports a novel hypothesis of strong association between metformin and intestinal immune system, and for the first time provide evidence for altered RNA expression as a contributing mechanism of metformin’s action. In addition to universal effects, 4 clusters of functionally related genes with a subject-specific differential expression were distinguished, including genes relevant to insulin production (HNF1B, HNF1A, HNF4A, GCK, INS, NEUROD1, PAX4, PDX1, ABCC8, KCNJ11) and cholesterol homeostasis (APOB, LDLR, PCSK9). This inter-individual variation of the metformin effect on the transcriptional regulation goes in line with well-known variability of the therapeutic response to the drug.publishersversionPeer reviewe

Shattered pellet injection experiments at JET in support of the ITER disruption mitigation system design

A series of experiments have been executed at JET to assess the efficacy of the newly installed shattered pellet injection (SPI) system in mitigating the effects of disruptions. Issues, important for the ITER disruption mitigation system, such as thermal load mitigation, avoidance of runaway electron (RE) formation, radiation asymmetries during thermal quench mitigation, electromagnetic load control and RE energy dissipation have been addressed over a large parameter range. The efficiency of the mitigation has been examined for the various SPI injection strategies. The paper summarises the results from these JET SPI experiments and discusses their implications for the ITER disruption mitigation scheme