Search results for: complex interventions

Authors: Clara Tramuta, Lucia Decastelli, Elisa Barcucci, Sandra Fragassi, Samantha Lupi, Enrico Arletti, Melissa Bizzarri, Daniela Manila Bianchi

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Introdution: Food allergies occurs, in the Western World, 2% of adults and up to 8% of children. The protection of allergic consumers is guaranted, in Eurrope, by Regulation (EU) No 1169/2011 of the European Parliament which governs the consumer's right to information and identifies 14 food allergens to be mandatory indicated on the label. Among these, mustard is a popular spice added to enhance the flavour and taste of foods. It is frequently present as an ingredient in spice blends, marinades, salad dressings, sausages, and other products. Hypersensitivity to mustard is a public health problem since the ingestion of even low amounts can trigger severe allergic reactions. In order to protect the allergic consumer, high performance methods are required for the detection of allergenic ingredients. Food safety laboratories rely on validated methods that detect hidden allergens in food to ensure the safety and health of allergic consumers. Here we present the test results for the validation and accreditation of a Real time PCR assay (RT-PCR: SPECIALfinder MC Mustard, Generon), for the detection of mustard traces in food. Materials and Methods. The method was tested on five classes of food matrices: bakery and pastry products (chocolate cookies), meats (ragù), ready-to-eat (mixed salad), dairy products (yogurt), grains, and milling products (rice and barley flour). Blank samples were spiked starting with the mustard samples (Sinapis Alba), lyophilized and stored at -18 °C, at a concentration of 1000 ppm. Serial dilutions were then prepared to a final concentration of 0.5 ppm, using the DNA extracted by ION Force FAST (Generon) from the blank samples. The Real Time PCR reaction was performed by RT-PCR SPECIALfinder MC Mustard (Generon), using CFX96 System (BioRad). Results. Real Time PCR showed a limit of detection (LOD) of 0.5 ppm in grains and milling products, ready-to-eat, meats, bakery, pastry products, and dairy products (range Ct 25-34). To determine the exclusivity parameter of the method, the ragù matrix was contaminated with Prunus dulcis (almonds), peanut (Arachis hypogaea), Glycine max (soy), Apium graveolens (celery), Allium cepa (onion), Pisum sativum (peas), Daucus carota (carrots), and Theobroma cacao (cocoa) and no cross-reactions were observed. Discussion. In terms of sensitivity, the Real Time PCR confirmed, even in complex matrix, a LOD of 0.5 ppm in five classes of food matrices tested; these values are compatible with the current regulatory situation that does not consider, at international level, to establish a quantitative criterion for the allergen considered in this study. The Real Time PCR SPECIALfinder kit for the detection of mustard proved to be easy to use and particularly appreciated for the rapid response times considering that the amplification and detection phase has a duration of less than 50 minutes. Method accuracy was rated satisfactory for sensitivity (100%) and specificity (100%) and was fully validated and accreditated. It was found adequate for the needs of the laboratory as it met the purpose for which it was applied. This study was funded in part within a project of the Italian Ministry of Health (IZS PLV 02/19 RC).

Keywords: allergens, food, mustard, real time PCR

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Authors: Ariesny Vera, Rodrigo Montecinos

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The rare-earth elements (REEs) or rare-earth metals (REMs), correspond to seventeen chemical elements composed by the fifteen lanthanoids, as well as scandium and yttrium. Lanthanoids corresponds to lanthanum and the f-block elements, from cerium to lutetium. Scandium and yttrium are considered rare-earth elements because they have ionic radii similar to the lighter f-block elements. These elements were called rare earths because they are simply more difficult to extract and separate individually than the most metals and, generally, they do not accumulate in minerals, they are rarely found in easily mined ores and are often unfavorably distributed in common ores/minerals. REEs show unique chemical and physical properties, in comparison to the other metals in the periodic table. Nowadays, these physicochemical properties are utilized in a wide range of synthetic, catalytic, electronic, medicinal, and military applications. Because of their applications, the global demand for rare earth metals is becoming progressively more important in the transition to a self-sustaining society and greener economy. However, due to the difficult separation between lanthanoid ions, the high cost and pollution of these processes, the scientists search the development of a method that combines selectivity and quantitative separation of lanthanoids from the leaching liquor, while being more economical and environmentally friendly processes. This motivation has favored the design and development of more efficient and environmentally friendly cation extractors with the incorporation of compounds as ionic liquids, membrane inclusion polymers (PIM) and supramolecular systems. Supramolecular chemistry focuses on the development of host-guest systems, in which a host molecule can recognize and bind a certain guest molecule or ion. Normally, the formation of a host-guest complex involves non-covalent interactions Additionally, host-guest interactions can be influenced among others effects by the structural nature of host and guests. The different macrocyclic hosts for lanthanoid species that have been studied are crown ethers, cyclodextrins, cucurbituryls, calixarenes and pillararenes.Among all the factors that can influence and affect lanthanoid (III) coordination, perhaps the most basic of them is the systematic control using macrocyclic substituents that promote a selective coordination. In this sense, macrocycles pillar[n]arenes (P[n]As) present a relatively easy functionalization and they have more π-rich cavity than other host molecules. This gives to P[n]As a negative electrostatic potential in the cavity which would be responsible for the selectivity of these compounds towards cations. Furthermore, the cavity size, the linker, and the functional groups of the polar headgroups could be modified in order to control the association of lanthanoid cations. In this sense, different P[n]As systems, specifically derivatives of the pentamer P[5]A functionalized with amide, amine, phosphate and sulfate derivatives, have been designed in terms of experimental synthesis and molecular dynamics, and the interaction between these P[5]As and some lanthanoid ions such as La³+, Eu³+ and Lu³+ has been studied by physicochemical characterization by 1H-NMR, ITC and fluorescence in the case of Eu³+ systems. The molecular dynamics study of these systems was developed in hexane as solvent, also taking into account the lanthanoid ions mentioned above, and the respective comparison studies between the different ions.

Keywords: lanthanoids, macrocycles, pillar[n]arenes, rare-earth metal extraction, supramolecular chemistry, supramolecular complexes.

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Authors: Pauline Nyssen, Ange Mouithys-Mickalad, Maryse Hoebeke

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Inflammation is a complex physiological phenomenon involving chemical and enzymatic mechanisms. Polymorphonuclear neutrophil leukocytes (PMNs) play an important role by producing reactive oxygen species (ROS) and releasing myeloperoxidase (MPO), a pro-oxidant enzyme. Released both in the phagolysosome and the extracellular medium, MPO produces during its peroxidase and halogenation cycles oxidant species, including hypochlorous acid, involved in the destruction of pathogen agents, like bacteria or viruses. Inflammatory pathologies, like rheumatoid arthritis, atherosclerosis induce an excessive stimulation of the PMNs and, therefore, an uncontrolled release of ROS and MPO in the extracellular medium, causing severe damages to the surrounding tissues and biomolecules such as proteins, lipids, and DNA. The treatment of chronic inflammatory pathologies remains a challenge. For many years, MPO has been used as a target for the development of effective treatments. Numerous studies have been focused on the design of new drugs presenting more efficient MPO inhibitory properties. However, some designed inhibitors can be toxic. An alternative consists of assessing the potential inhibitory action of clinically-known molecules, having antioxidant activity. Propofol, 2,6-diisopropyl phenol, which is used as an intravenous anesthetic agent, meets these requirements. Besides its anesthetic action employed to induce a sedative state during surgery or in intensive care units, propofol and its injectable form Diprivan indeed present antioxidant properties and act as ROS and free radical scavengers. A study has also evidenced the ability of propofol to inhibit the formation of the neutrophil extracellular traps fibers, which are important to trap pathogen microorganisms during the inflammation process. The aim of this study was to investigate the potential inhibitory action mechanism of propofol and Diprivan on MPO activity. To go into the anti-inflammatory action of propofol in-depth, two of its oxidative derivatives, 2,6-diisopropyl-1,4-p-benzoquinone (PPFQ) and 3,5,3’,5’-tetra isopropyl-(4,4’)-diphenoquinone (PPFDQ), were studied regarding their inhibitory action. Specific immunological extraction followed by enzyme detection (SIEFED) and molecular modeling have evidenced the low anti-catalytic action of propofol. Stopped-flow absorption spectroscopy and direct MPO activity analysis have proved that propofol acts as a reversible MPO inhibitor by interacting as a reductive substrate in the peroxidase cycle and promoting the accumulation of redox compound II. Overall, Diprivan exhibited a weaker inhibitory action than the active molecule propofol. In contrast, PPFQ seemed to bind and obstruct the enzyme active site, preventing the trigger of the MPO oxidant cycles. PPFQ induced a better chlorination cycle inhibition at basic and neutral pH in comparison to propofol. PPFDQ did not show any MPO inhibition activity. The three interest molecules have also demonstrated their inhibition ability on an important step of the inflammation pathway, the PMNs superoxide anions production, thanks to EPR spectroscopy and chemiluminescence. In conclusion, propofol presents an interesting immunomodulatory activity by acting as a reductive substrate in the peroxidase cycle of MPO, slowing down its activity, whereas PPFQ acts more as an anti-catalytic substrate. Although PPFDQ has no impact on MPO, it can act on the inflammation process by inhibiting the superoxide anions production by PMNs.

Keywords: Diprivan, inhibitor, myeloperoxidase, propofol, spectroscopy

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Authors: Ana Fonseca, Ana Gouveia, Edgar Ramalho, Rita Henriques, Filipa Figueiredo, João Nunes

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Due to persistent climate pressures, there is a need to find a resilient economic system that is regenerative in nature. Bioeconomy offers the possibility of replacing non-renewable and non-biodegradable materials derived from fossil fuels with ones that are renewable and biodegradable, while a Circular Economy aims at sustainable and resource-efficient operations. The term "Circular Bioeconomy", which can be summarized as all activities that transform biomass for its use in various product streams, expresses the interaction between these two ideas. Portugal has a very favourable context to promote a Circular Bioeconomy due to its variety of climates and ecosystems, availability of biologically based resources, location, and geomorphology. Recently, there have been political and legislative efforts to develop the Portuguese Circular Bioeconomy. The Action Plan for a Sustainable Bioeconomy, approved in 2021, is composed of five axes of intervention, ranging from sustainable production and the use of regionally based biological resources to the development of a circular and sustainable bioindustry through research and innovation. However, as some statistics show, Portugal is still far from achieving circularity. According to Eurostat, Portugal has circularity rates of 2.8%, which is the second lowest among the member states of the European Union. Some challenges contribute to this scenario, including sectorial heterogeneity and fragmentation, prevalence of small producers, lack of attractiveness for younger generations, and absence of implementation of collaborative solutions amongst producers and along value chains.Regarding the Portuguese industrial sector, there is a tendency towards complex bureaucratic processes, which leads to economic and financial obstacles and an unclear national strategy. Together with the limited number of incentives the country has to offer to those that pretend to abandon the linear economic model, many entrepreneurs are hesitant to invest the capital needed to make their companies more circular. Absence of disaggregated, georeferenced, and reliable information regarding the actual availability of biological resources is also a major issue. Low literacy on bioeconomy among many of the sectoral agents and in society in general directly impacts the decisions of production and final consumption. The WinBio project seeks to outline a strategic approach for the management of weaknesses/opportunities in the technology transfer process, given the reality of the territory, through road mapping and national and international benchmarking. The developed work included the identification and analysis of agents in the interior region of Portugal, natural endogenous resources, products, and processes associated with potential development. Specific flow of biological wastes, possible value chains, and the potential for replacing critical raw materials with bio-based products was accessed, taking into consideration other countries with a matured bioeconomy. The study found food industry, agriculture, forestry, and fisheries generate huge amounts of waste streams, which in turn provide an opportunity for the establishment of local bio-industries powered by this biomass. The project identified biological resources with potential for replication and applicability in the Portuguese context. The richness of natural resources and potentials known in the interior region of Portugal is a major key to developing the Circular Economy and sustainability of the country.

Keywords: circular bioeconomy, interior region of portugal, regional development., public policy

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Authors: Claudio Loconsole, Daniele Leonardis, Antonio Brunetti, Gianpaolo Francesco Trotta, Nicholas Caporusso, Vitoantonio Bevilacqua

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Unfortunately, many people still do not have access to communication, with specific regard to reading and writing. Among them, people who are blind or visually impaired, have several difficulties in getting access to the world, compared to the sighted. Indeed, despite technology advancement and cost reduction, nowadays assistive devices are still expensive such as Braille-based input/output systems which enable reading and writing texts (e.g., personal notes, documents). As a consequence, assistive technology affordability is fundamental in supporting the visually impaired in communication, learning, and social inclusion. This, in turn, has serious consequences in terms of equal access to opportunities, freedom of expression, and actual and independent participation to a society designed for the sighted. Moreover, the visually impaired experience difficulties in recognizing objects and interacting with devices in any activities of daily living. It is not a case that Braille indications are commonly reported only on medicine boxes and elevator keypads. Several software applications for the automatic translation of written text into speech (e.g., Text-To-Speech - TTS) enable reading pieces of documents. However, apart from simple tasks, in many circumstances TTS software is not suitable for understanding very complicated pieces of text requiring to dwell more on specific portions (e.g., mathematical formulas or Greek text). In addition, the experience of reading\writing text is completely different both in terms of engagement, and from an educational perspective. Statistics on the employment rate of blind people show that learning to read and write provides the visually impaired with up to 80% more opportunities of finding a job. Especially in higher educational levels, where the ability to digest very complex text is key, accessibility and availability of Braille plays a fundamental role in reducing drop-out rate of the visually impaired, thus affecting the effectiveness of the constitutional right to get access to education. In this context, the Braille Lab project aims at overcoming these social needs by including affordability in designing and developing assistive tools for visually impaired people. In detail, our awarded project focuses on a technology innovation of the operation principle of existing assistive tools for the visually impaired leaving the Human-Machine Interface unchanged. This can result in a significant reduction of the production costs and consequently of tool selling prices, thus representing an important opportunity for social entrepreneurship. The first two assistive tools designed within the Braille Lab project following the proposed approach aims to provide the possibility to personally print documents and handouts and to read texts written in Braille using refreshable Braille display, respectively. The former, named ‘Braille Cartridge’, represents an alternative solution for printing in Braille and consists in the realization of an electronic-controlled dispenser printing (cartridge) which can be integrated within traditional ink-jet printers, in order to leverage the efficiency and cost of the device mechanical structure which are already being used. The latter, named ‘Braille Cursor’, is an innovative Braille display featuring a substantial technology innovation by means of a unique cursor virtualizing Braille cells, thus limiting the number of active pins needed for Braille characters.

Keywords: Human rights, social challenges and technology innovations, visually impaired, affordability, assistive tools

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Authors: Hasneet K. Kalsi, Julie K. Kilgariff

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Introduction: Within dentistry, the process for gaining informed consent has become more complex. To consent for treatment, patients must understand all reasonable treatment options and associated risks and benefits. Consenting is therefore deeply embedded in health literacy. Patients attending for dental consultation are often presented with an array of information and choices, yet studies show patients recall less than half of the information provided immediately after. Appropriate and comprehensible patient information leaflets (PILs) may be useful aid memories. In 2016 the World Health Organisation set improving health literacy as a global priority. Soon after, Scotland’s 2017-2025 Making it Easier: A Health Literacy Action Plan followed. This project involved the review of Restorative PILs used within Dundee Dental Hospital to assess the Content and Readability. Method: The current PIL on Root Canal Treatment (RCT) was created in 2011. This predates the Montgomery vs. NHS Lanarkshire case, a ruling which significantly impacted dental consenting processes, as well as General Dental Council’s (GDC’s) Standards for the Dental Team and Faculty of General Dental Practice’s Good Practice Guidance on Clinical Examination and Record-Keeping. Current evidence-based guidance, including that stipulated by the GDC, was reviewed. A 20-point Essential Content Checklist was designed to conform to best practice guidance for valid consenting processes. The RCT leaflet was scored against this to ascertain if the content was satisfactory. Having ensured the content satisfied medicolegal requirements, health literacy considerations were reviewed regarding readability. This was assessed using McLaughlin’s Simple Measure of Gobbledygook (SMOG) formula, which identifies school stages that would have to be achieved to comprehend the PIL. The sensitivity of the results to alternative readability methods were assessed. Results: The PIL was not sufficient for modern consenting processes and reflected a suboptimal level of health literacy. Evaluation of the leaflet revealed key content was missing, including information pertaining to risks and benefits. Only five points out of the 20-point checklist were present. The readability score was 16, equivalent to a level 2 in National Adult Literacy Standards/Scottish Credit and Qualification Framework Level 5; 62% of Scottish adults are able to read to this standard. Discussion: Assessment of the leaflet showed it was no longer fit for purpose. Reasons include a lack of pertinent information, a text-heavy leaflet lacking flow, and content errors. The SMOG score indicates a high level of comprehension is required to understand this PIL, which many patients may not possess. A new PIL, compliant with medicolegal and health literacy guidance, was designed with patient-driven checklists, notes spaces for annotations/ questions and areas for clinicians to highlight important case-specific information. It has been tested using the SMOG formula. Conclusion: PILs can be extremely useful. Studies show that interactive use can enhance their effectiveness. PILs should reflect best practice guidance and be understood by patients. The 2020 leaflet designed and implemented aims to fulfill the needs of a modern healthcare system and its service users. It embraces and embeds Scotland’s Health Literacy Action Plan within the consenting process. A review of further leaflets using this model is ongoing.

Keywords: consent, health literacy, patient information leaflet, restorative dentistry

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Authors: Michelle Maurer, Antonia Last, Mark S. Gresnigt, Bernhard Hube, Alexander S. Mosig

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The intestinal epithelium forms an essential barrier to prevent translocation of microorganisms, toxins or other potentially harmful molecules into the bloodstream. In particular, dendritic cells of the intestinal epithelium orchestrate an adapted response of immune tolerance to commensals and immune defense against invading pathogens. Systemic inflammation is typically associated with a dysregulation of this adapted immune response and is accompanied by a disruption of the epithelial and endothelial gut barrier which enables dissemination of pathogens within the human body. To understand the pathophysiological mechanisms underlying the inflammation-associated gut barrier breakdown, it is crucial to elucidate the complex interplay of the host and the intestinal microbiome. A microfluidically perfused three-dimensional intestine-on-chip model was established to emulate these processes in the presence of immune cells, commensal bacteria, and facultative pathogens. Multi-organ tissue flow (MOTiF) biochips made from polystyrene were used for microfluidic perfusion of the intestinal tissue model. The biochips are composed of two chambers separated by a microporous membrane. Each chamber is connected to inlet and outlet channels allowing independent perfusion of the individual channels and application of microfluidic shear stress. Human umbilical vein endothelial cells (HUVECs), monocyte-derived macrophages and intestinal epithelial cells (Caco-2) were assembled on the biochip membrane. Following 7 – 14 days of growth in the presence of physiological flow conditions, the epithelium was colonized with the commensal bacterium Lactobacillus rhamnosus, while the endothelium was perfused with peripheral blood mononuclear cells (PBMCs). Additionally, L. rhamnosus was co-cultivated with the opportunistic fungal pathogen Candida albicans. Within one week of perfusion, the epithelial cells formed self-organized and well-polarized villus- and crypt-like structures that resemble essential morphological characteristics of the human intestine. Dendritic cells were differentiated in the epithelial tissue that specifically responds to bacterial lipopolysaccharide (LPS) challenge. LPS is well-tolerated at the luminal epithelial side of the intestinal model without signs of tissue damage or induction of an inflammatory response, even in the presence of circulating PBMC at the endothelial lining. In contrast, LPS stimulation at the endothelial side of the intestinal model triggered the release of pro-inflammatory cytokines such as TNF, IL-1β, IL-6, and IL-8 via activation of macrophages residing in the endothelium. Perfusion of the endothelium with PBMCs led to an enhanced cytokine release. L. rhamnosus colonization of the model was tolerated in the immune competent tissue model and was demonstrated to reduce damage induced by C. albicans infection. A microfluidic intestine-on-chip model was developed to mimic a systemic infection with a dysregulated immune response under physiological conditions. The model facilitates the colonization of commensal bacteria and co-cultivation with facultative pathogenic microorganisms. Both, commensal bacteria alone and facultative pathogens controlled by commensals, are tolerated by the host and contribute to cell signaling. The human intestine-on-chip model represents a promising tool to mimic microphysiological conditions of the human intestine and paves the way for more detailed in vitro studies of host-microbiota interactions under physiologically relevant conditions.

Keywords: host-microbiota interaction, immune tolerance, microfluidics, organ-on-chip

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Authors: Yunwei Tang, Linhai Jing, Hui Li, Qingjie Liu, Xiuxia Li, Qi Yan, Haifeng Ding

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The giant panda (Ailuropoda melanoleuca) is an endangered species, mainly live in central China, where bamboos act as the main food source of wild giant pandas. Knowledge of spatial distribution of bamboos therefore becomes important for identifying the habitat of giant pandas. There have been ongoing studies for mapping bamboos and other tree species using remote sensing. WorldView-2 (WV-2) is the first high resolution commercial satellite with eight Multi-Spectral (MS) bands. Recent studies demonstrated that WV-2 imagery has a high potential in classification of tree species. The advanced classification techniques are important for utilising high spatial resolution imagery. It is generally agreed that object-based image analysis is a more desirable method than pixel-based analysis in processing high spatial resolution remotely sensed data. Classifiers that use spatial information combined with spectral information are known as contextual classifiers. It is suggested that contextual classifiers can achieve greater accuracy than non-contextual classifiers. Thus, spatial correlation can be incorporated into classifiers to improve classification results. The study area is located at Wuyipeng area in Wolong, Sichuan Province. The complex environment makes it difficult for information extraction since bamboos are sparsely distributed, mixed with brushes, and covered by other trees. Extensive fieldworks in Wuyingpeng were carried out twice. The first one was on 11th June, 2014, aiming at sampling feature locations for geometric correction and collecting training samples for classification. The second fieldwork was on 11th September, 2014, for the purposes of testing the classification results. In this study, spectral separability analysis was first performed to select appropriate MS bands for classification. Also, the reflectance analysis provided information for expanding sample points under the circumstance of knowing only a few. Then, a spatially weighted object-based k-nearest neighbour (k-NN) classifier was applied to the selected MS bands to identify seven land cover types (bamboo, conifer, broadleaf, mixed forest, brush, bare land, and shadow), accounting for spatial correlation within classes using geostatistical modelling. The spatially weighted k-NN method was compared with three alternatives: the traditional k-NN classifier, the Support Vector Machine (SVM) method and the Classification and Regression Tree (CART). Through field validation, it was proved that the classification result obtained using the spatially weighted k-NN method has the highest overall classification accuracy (77.61%) and Kappa coefficient (0.729); the producer’s accuracy and user’s accuracy achieve 81.25% and 95.12% for the bamboo class, respectively, also higher than the other methods. Photos of tree crowns were taken at sample locations using a fisheye camera, so the canopy density could be estimated. It is found that it is difficult to identify bamboo in the areas with a large canopy density (over 0.70); it is possible to extract bamboos in the areas with a median canopy density (from 0.2 to 0.7) and in a sparse forest (canopy density is less than 0.2). In summary, this study explores the ability of WV-2 imagery for bamboo extraction in a mountainous region in Sichuan. The study successfully identified the bamboo distribution, providing supporting knowledge for assessing the habitats of giant pandas.

Keywords: bamboo mapping, classification, geostatistics, k-NN, worldview-2

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Authors: Danyi Wang, Andrew Schriefer, Dennis O'Rourke, Brajendra Kumar, Yang Liu, Fei Zhong, Juergen Scheuenpflug, Zheng Feng

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Purpose: It has been recognized that the microbiome plays critical roles in disease pathogenesis, including cancer, autoimmune disease, and multiple sclerosis. To develop a clinical-grade assay for exploring microbiome-derived clinical biomarkers across disease areas, a two-phase approach is implemented. 1) Identification of the optimal sample preparation reagents using pre-mixed bacteria and healthy donor stool samples coupled with proprietary Sigma-Aldrich® bioinformatics solution. 2) Exploratory analysis of patient samples for enabling precision medicine. Study Procedure: In phase 1 study, we first compared the 16S sequencing results of two ATCC® microbiome standards (MSA 2002 and MSA 2003) across five different extraction kits (Kit A, B, C, D & E). Both microbiome standards samples were extracted in triplicate across all extraction kits. Following isolation, DNA quantity was determined by Qubit assay. DNA quality was assessed to determine purity and to confirm extracted DNA is of high molecular weight. Bacterial 16S ribosomal ribonucleic acid (rRNA) amplicons were generated via amplification of the V3/V4 hypervariable region of the 16S rRNA. Sequencing was performed using a 2x300 bp paired-end configuration on the Illumina MiSeq. Fastq files were analyzed using the Sigma-Aldrich® Microbiome Platform. The Microbiome Platform is a cloud-based service that offers best-in-class 16S-seq and WGS analysis pipelines and databases. The Platform and its methods have been extensively benchmarked using microbiome standards generated internally by MilliporeSigma and other external providers. Data Summary: The DNA yield using the extraction kit D and E is below the limit of detection (100 pg/µl) of Qubit assay as both extraction kits are intended for samples with low bacterial counts. The pre-mixed bacterial pellets at high concentrations with an input of 2 x106 cells for MSA-2002 and 1 x106 cells from MSA-2003 were not compatible with the kits. Among the remaining 3 extraction kits, kit A produced the greatest yield whereas kit B provided the least yield (Kit-A/MSA-2002: 174.25 ± 34.98; Kit-A/MSA-2003: 179.89 ± 30.18; Kit-B/MSA-2002: 27.86 ± 9.35; Kit-B/MSA-2003: 23.14 ± 6.39; Kit-C/MSA-2002: 55.19 ± 10.18; Kit-C/MSA-2003: 35.80 ± 11.41 (Mean ± SD)). Also, kit A produced the greatest yield, whereas kit B provided the least yield. The PCoA 3D visualization of the Weighted Unifrac beta diversity shows that kits A and C cluster closely together while kit B appears as an outlier. The kit A sequencing samples cluster more closely together than both the other kits. The taxonomic profiles of kit B have lower recall when compared to the known mixture profiles indicating that kit B was inefficient at detecting some of the bacteria. Conclusion: Our data demonstrated that the DNA extraction method impacts DNA concentration, purity, and microbial communities detected by next-generation sequencing analysis. Further microbiome analysis performance comparison of using healthy stool samples is underway; also, colorectal cancer patients' samples will be acquired for further explore the clinical utilities. Collectively, our comprehensive qualification approach, including the evaluation of optimal DNA extraction conditions, the inclusion of positive controls, and the implementation of a robust qualified bioinformatics pipeline, assures accurate characterization of the microbiota in a complex matrix for deciphering the deep biology and enabling precision medicine.

Keywords: 16S rRNA sequencing, analytical validation, bioinformatics pipeline, metagenomics

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Authors: Batul Kagalwala

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The nervous system is made up of complex delicate structures such as the spinal cord, peripheral nerves and the brain. These are prone to various types of injury ranging from neurodegenerative diseases to trauma leading to diseases like Parkinson's, Alzheimer's, multiple sclerosis, amyotrophic lateral sclerosis (ALS), multiple system atrophy etc. Unfortunately, because of the complicated structure of nervous system, spontaneous regeneration, repair and healing is seldom seen due to which brain damage, peripheral nerve damage and paralysis from spinal cord injury are often permanent and incapacitating. Hence, innovative and standardized approach is required for advance treatment of neurological injury. Nigella sativa (N. sativa), an annual flowering plant native to regions of southern Europe and Asia; has been suggested to have neuroprotective and anti-seizures properties. Neuroregeneration is found to occur in damaged cells when treated using extract of N. sativa. Due to its proven health benefits, lots of experiments are being conducted to extract all the benefits from the plant. The flowers are delicate and are usually pale blue and white in color with small black seeds. These seeds are the source of active components such as 30–40% fixed oils, 0.5–1.5% essential oils, pharmacologically active components containing thymoquinone (TQ), ditimoquinone (DTQ) and nigellin. In traditional medicine, this herb was identified to have healing properties and was extensively used Middle East and Far East for treating diseases such as head ache, back pain, asthma, infections, dysentery, hypertension, obesity and gastrointestinal problems. Literature studies have confirmed the extract of N. sativa seeds and TQ have inhibitory effects on inducible nitric oxide synthase and production of nitric oxide as well as anti-inflammatory and anticancer activities. Experimental investigation will be conducted to understand which ingredient of N. sativa causes neuroregeneration and roots to its healing property. An aqueous/ alcoholic extract of N. sativa will be made. Seed oil is also found to have used by researchers to prepare such extracts. For the alcoholic extracts, the seeds need to be powdered and soaked in alcohol for a period of time and the alcohol must be evaporated using rotary evaporator. For aqueous extracts, the powder must be dissolved in distilled water to obtain a pure extract. The mobile phase will be the extract while the suitable stationary phase (substance that is a good adsorbent e.g. silica gels, alumina, cellulose etc.) will be selected. Different ingredients of N. sativa will be separated using High Performance Liquid Chromatography (HPLC) for treating damaged cells. Damaged brain cells will be treated individually and in different combinations of 2 or 3 compounds for different intervals of time. The most suitable compound or a combination of compounds for the regeneration of cells will be determined using DOE methodology. Later the gene will also be determined and using Polymerase Chain Reaction (PCR) it will be replicated in a plasmid vector. This plasmid vector shall be inserted in the brain of the organism used and replicated within. The gene insertion can also be done by the gene gun method. The gene in question can be coated on a micro bullet of tungsten and bombarded in the area of interest and gene replication and coding shall be studied. Investigation on whether the gene replicates in the organism or not will be examined.

Keywords: black cumin, brain cells, damage, extract, neuroregeneration, PCR, plasmids, vectors

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Authors: D. Malouch, M. Berchel, C. Dreanno, S. Stachowski-Haberkorn, P-A. Jaffres

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Biofilm is a predominant lifestyle chosen by bacteria. Whether it is developed on an immerged surface or a mobile biofilm known as flocs, the bacteria within this form of life show properties different from its planktonic ones. Within the biofilm, the self-formed matrix of Extracellular Polymeric Substances (EPS) offers hydration, resources capture, enhanced resistance to antimicrobial agents, and allows cell-communication. Biofouling is a complex natural phenomenon that involves biological, physical and chemical properties related to the environment, the submerged surface and the living organisms involved. Bio-colonization of artificial structures can cause various economic and environmental impacts. The increase in costs associated with the over-consumption of fuel from biocolonized vessels has been widely studied. Measurement drifts from submerged sensors, as well as obstructions in heat exchangers, and deterioration of offshore structures are major difficulties that industries are dealing with. Therefore, surfaces that inhibit biocolonization are required in different areas (water treatment, marine paints, etc.) and many efforts have been devoted to produce efficient and eco-compatible antifouling agents. The different steps of surface fouling are widely described in literature. Studying the biofilm and its stages provides a better understanding of how to elaborate more efficient antifouling strategies. Several approaches are currently applied, such as the use of biocide anti-fouling paint6 (mainly with copper derivatives) and super-hydrophobic coatings. While these two processes are proving to be the most effective, they are not entirely satisfactory, especially in a context of a changing legislation. Nowadays, the challenge is to prevent biofouling with non-biocide compounds, offering a cost effective solution, but with no toxic effects on marine organisms. Since the micro-fouling phase plays an important role in the regulation of the following steps of biofilm formation7, it is desired to reduce or delate biofouling of a given surface by inhibiting the micro fouling at its early stages. In our recent works, we reported that some amphiphilic compounds exhibited bacteriostatic or bactericidal properties at a concentration that did not affect eukaryotic cells. These remarkable properties invited us to assess this type of bio-inspired phospholipids9 to prevent the colonization of surfaces by marine bacteria. Of note, other studies reported that amphiphilic compounds interacted with bacteria leading to a reduction of their development. An amphiphilic compound is a molecule consisting of a hydrophobic domain and a polar head (ionic or non-ionic). These compounds appear to have interesting antifouling properties: some ionic compounds have shown antimicrobial activity, and zwitterions can reduce nonspecific adsorption of proteins. Herein, we investigate the potential of amphiphilic compounds as inhibitors of bacterial growth and marine biofilm formation. The aim of this study is to compare the efficacy of four synthetic phospholipids that features a cationic charge (BSV36, KLN47) or a zwitterionic polar-head group (SL386, MB2871) to prevent microfouling with marine bacteria. We also study the toxicity of these compounds in order to identify the most promising compound that must feature high anti-adhesive properties and a low cytotoxicity on two links representative of coastal marine food webs: phytoplankton and oyster larvae.

Keywords: amphiphilic phospholipids, bacterial biofilm, marine microfouling, non-toxic antifouling

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Authors: Mahbuba Kaneez Hasna

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Introductory Statement outlining the background: Little has been written about political ecology of urban gardening, such as a network of knowledge generation, technologies of food production and distribution, food consumption practices, and the regulation of ‘agricultural activities. For urban food gardens to sustain as a long-term food security enterprise, we will need to better understand the anthropological, ecological, political, and institutional factors influencing their development, management, and ongoing viability. Significance of the study: Dhaka as one of the fastest growing city. There are currently no studies regards to Bangladesh on how urban slum dwellerscope with the changing urban environment in the city, where they overcome challenges, and how they cope with the urban ecological cycle of food and vegetable production. It is also essential to understand the importance of their access to confined spaces in the slums they apply their indigenous knowledge. These relationships in nature are important factors in community and conservation ecology. Until now, there has been no significant published academic work on relationships between urban and environmental anthropology, urban planning, geography, ecology, and social anthropology with a focus on urban agriculture and how this contributes to the moral economies, indigenous knowledge, and government policies in order to improve the lives and livelihoods of slum dwellers surrounding parks and open spaces in Dhaka, Bangladesh. Methodology: it have applied participant observation, semi-structured questionnaire-based interviews, and focus group discussions to collect social data. Interviews were conducted with the urban agriculture practitioners who are slum dwellers who carry out their urban agriculture activities. Some of the interviews were conducted with non-government organisations (NGOs) and local and state government officials, using semi-structured interviews. Using these methods developed a clearer understanding of how green space cultivation, local economic self-reliance, and urban gardening are producing distinctive urban ecologies in Dhaka and their policy-implications on urban sustainability. Major findings of the study: The research provided an in-depth knowledge on the challenges that slum dwellers encounter in establishing and maintaining urban gardens, such as the economic development of the city, conflicting political agendas, and environmental constraints in areas within which gardening activities take place. The research investigated (i) How do slum dwellers perform gardening practices from rural areas to open spaces in the city? (ii) How do men and women’s ethno-botanical knowledge contribute to urban biodiversity; (iii) And how do slum dwellers navigate complex constellations of land use policy, competing political agendas, and conflicting land and water tenures to meet livelihood functions provided by their gardens. Concluding statement: Lack of infrastructure facilities such as water supply and sanitation, micro-drains and waste disposal areas, and poor access to basic health care services increase the misery of people in the slum areas. Lack of environmental health awareness information for farmers, such as the risks from the use of chemical pesticides in gardens and from grazing animals in contaminated fields or cropping and planting trees or vegetable in contaminated dumping grounds, can all cause high health risk to humans and their environment.

Keywords: gender, urban agriculture, ecosystem health, urban slum systems

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Authors: Susana Barreto

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This paper explores a pedagogical exercise that employs family photos as catalysts for teaching visual analysis and inspiring academic writing among MA students. The study aimed to achieve two primary objectives: to impart students with the skills of analyzing images or artifacts and to ignite their writing for research purposes. Conducted at Viana Polytechnic in Portugal, the exercise involved two classes on Arts Management and Art Education Master course comprising approximately twenty students from diverse academic backgrounds, including Economics, Design, Fine Arts, and Sociology, among others. The exploratory exercise involved selecting an old family photo, analyzing its content and context, and deconstructing the chosen images in an intuitive and systematic manner. Students were encouraged to engage in photo elicitation, seeking insights from family/friends to gain multigenerational perspectives on the images. The feedback received from this exercise was consistently positive, largely due to the personal connection students felt with the objects of analysis. Family photos, with their emotional significance, fostered deeper engagement and motivation in the learning process. Furthermore, visual analysing family photos stimulated critical thinking as students interpreted the composition, subject matter, and potential meanings embedded in the images. This practice enhanced their ability to comprehend complex visual representations and construct compelling visual narratives, thereby facilitating the writing process. The exercise also facilitated the identification of patterns, similarities, and differences by comparing different family photos, leading to a more comprehensive analysis of visual elements and themes. Throughout the exercise, students found analyzing their own photographs both enjoyable and insightful. They progressed through preliminary analysis, explored content and context, and artfully interwove these components. Additionally, students experimented with various techniques such as converting photos to black and white, altering framing angles, and adjusting sizes to unveil hidden meanings.The methodology employed included observation, documental analysis of written reports, and student interviews. By including students from diverse academic backgrounds, the study enhanced its external validity, enabling a broader range of perspectives and insights during the exercise. Furthermore, encouraging students to seek multigenerational perspectives from family and friends added depth to the analysis, enriching the learning experience and broadening the understanding of the cultural and historical context associated with the family photos Highlighting the emotional significance of these family photos and the personal connection students felt with the objects of analysis fosters a deeper connection to the subject matter. Moreover, the emphasis on stimulating critical thinking through the analysis of composition, subject matter, and potential meanings in family photos suggests a targeted approach to developing analytical skills. This improvement focuses specifically on critical thinking and visual analysis, enhancing the overall quality of the exercise. Additionally, the inclusion of a step where students compare different family photos to identify patterns, similarities, and differences further enhances the depth of the analysis. This comparative approach adds a layer of complexity to the exercise, ultimately leading to a more comprehensive understanding of visual elements and themes. The expected results of this study will culminate in a set of practical recommendations for implementing this exercise in academic settings.

Keywords: visual analysis, academic writing, pedagogical exercise, family photos

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Authors: Rachel Danielson, Brendan Bohannan, S.M. Tsai, Kyle Meyer, Jorge L.M. Rodrigues

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The Amazon Rainforest is a global diversity hotspot and crucial carbon sink, but approximately 20% of its total extent has been deforested- primarily for the establishment of cattle pasture. Understanding the impact of this large-scale disturbance on soil microbial community composition and activity is crucial in understanding potentially consequential shifts in nutrient or greenhouse gas cycling, as well as adding to the body of knowledge concerning how these complex communities respond to human disturbance. In this study, surface soils (0-10cm) were collected from three forests and three 45-year-old pastures in Rondonia, Brazil (the Amazon state with the greatest rate of forest destruction) in order to determine the impact of forest conversion on microbial communities involved in nitrogen fixation. Soil chemical and physical parameters were paired with measurements of microbial activity and genetic profiles to determine how community composition and process rates relate to environmental conditions. Measuring both the natural abundance of 15N in total soil N, as well as incorporation of enriched 15N2 under incubation has revealed that conversion of primary forest to cattle pasture results in a significant increase in the rate of nitrogen fixation by free-living diazotrophs. Quantification of nifH gene copy numbers (an essential subunit encoding the nitrogenase enzyme) correspondingly reveals a significant increase of genes in pasture compared to forest soils. Additionally, genetic sequencing of both nifH genes and transcripts shows a significant increase in the diversity of the present and metabolically active diazotrophs within the soil community. Levels of both organic and inorganic nitrogen tend to be lower in pastures compared to forests, with ammonium rather than nitrate as the dominant inorganic form. However, no significant or consistent differences in total, extractable, permanganate-oxidizable, or loss-on-ignition carbon are present between the two land-use types. Forest conversion is associated with a 0.5- 1.0 unit pH increase, but concentrations of many biologically relevant nutrients such as phosphorus do not increase consistently. Increases in free-living diazotrophic community abundance and activity appear to be related to shifts in carbon to nitrogen pool ratios. Furthermore, there may be an important impact of transient, low molecular weight plant-root-derived organic carbon on free-living diazotroph communities not captured in this study. Preliminary analysis of nitrogenase gene variant composition using NovoSeq metagenomic sequencing indicates that conversion of forest to pasture may significantly enrich vanadium-based nitrogenases. This indication is complemented by a significant decrease in available soil molybdenum. Very little is known about the ecology of diazotrophs utilizing vanadium-based nitrogenases, so further analysis may reveal important environmental conditions favoring their abundance and diversity in soil systems. Taken together, the results of this study indicate a significant change in nitrogen cycling and diazotroph community composition with the conversion of the Amazon Rainforest. This may have important implications for the sustainability of cattle pastures once established since nitrogen is a crucial nutrient for forage grass productivity.

Keywords: free-living diazotrophs, land use change, metagenomic sequencing, nitrogen fixation

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Authors: Camille Perréard, Yoann Ladner, Fanny D'Orlyé, Stéphanie Descroix, Vélan Taniga, Anne Varenne, Cédric Guyon, Michael. Tatoulian, Frédéric Kanoufi, Cyrine Slim, Sophie Griveau, Fethi Bedioui

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The development of micro total analysis systems is of major interest for contaminant and biomarker analysis. As a lab-on-chip integrates all steps of an analysis procedure in a single device, analysis can be performed in an automated format with reduced time and cost, while maintaining performances comparable to those of conventional chromatographic systems. Moreover, these miniaturized systems are either compatible with field work or glovebox manipulations. This work is aimed at developing an analytical microsystem for trace and ultra trace quantitation in complex matrices. The strategy consists in the integration of a sample pretreatment step within the lab-on-chip by a confinement zone where selective ligands are immobilized for target extraction and preconcentration. Aptamers were chosen as selective ligands, because of their high affinity for all types of targets (from small ions to viruses and cells) and their ease of synthesis and functionalization. This integrated target extraction and concentration step will be followed in the microdevice by an electrokinetic separation step and an on-line detection. Polymers consisting of cyclic olefin copolymer (COC) or fluoropolymer (Dyneon THV) were selected as they are easy to mold, transparent in UV-visible and have high resistance towards solvents and extreme pH conditions. However, because of their low chemical reactivity, surface treatments are necessary. For the design of this miniaturized diagnostics, we aimed at modifying the microfluidic system at two scales : (1) on the entire surface of the microsystem to control the surface hydrophobicity (so as to avoid any sample wall adsorption) and the fluid flows during electrokinetic separation, or (2) locally so as to immobilize selective ligands (aptamers) on restricted areas for target extraction and preconcentration. We developed different novel strategies for the surface functionalization of COC and Dyneon, based on plasma, chemical and /or electrochemical approaches. In a first approach, a plasma-induced immobilization of brominated derivatives was performed on the entire surface. Further substitution of the bromine by an azide functional group led to covalent immobilization of ligands through “click” chemistry reaction between azides and terminal alkynes. COC and Dyneon materials were characterized at each step of the surface functionalization procedure by various complementary techniques to evaluate the quality and homogeneity of the functionalization (contact angle, XPS, ATR). With the objective of local (micrometric scale) aptamer immobilization, we developed an original electrochemical strategy on engraved Dyneon THV microchannel. Through local electrochemical carbonization followed by adsorption of azide-bearing diazonium moieties and covalent linkage of alkyne-bearing aptamers through click chemistry reaction, typical dimensions of immobilization zones reached the 50 µm range. Other functionalization strategies, such as sol-gel encapsulation of aptamers, are currently investigated and may also be suitable for the development of the analytical microdevice. The development of these functionalization strategies is the first crucial step in the design of the entire microdevice. These strategies allow the grafting of a large number of molecules for the development of new analytical tools in various domains like environment or healthcare.

Keywords: alkyne-azide click chemistry (CuAAC), electrochemical modification, microsystem, plasma bromination, surface functionalization, thermoplastic polymers

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Authors: Aparajita Bhattacharya, Mousumi Dutta, Zakir Husain, Dionne Sequeira, Asima Mukhopadhyay

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Background: Tata Medical Centre (TMC), Kolkata is a major cancer referral centre in Eastern India and neighbouring countries providing state of the art facilities; however, it is a non-profit organisation with patients requiring to pay at subsidised rates. Although a system for social assessment and applying for governmental/ non-governmental (NGO) funds is in place, access is challenging. Amongst gynaecological cancers (GC), ovarian cancer (OC) is associated with the highest treatment cost; majority of which is required at the beginning when complex surgery is performed and funding arrangements cannot be made in time. We therefore appointed a dedicated Medical Social Worker (MSW) in 2016, supported by NGO for GC patients in order to assist patients/family members to access/avail these funds more readily and assist in economic evaluation for both direct and opportunity costs. Objectives: To reflect on our experience and challenges in collecting data on economic evaluation of cancer patients and compare success rates in achieving fund mobilization after introduction of MSW. Methods: A Retrospective survey. Patients with OC and their relatives were seen by the MSW during the initial outpatients department visit and followed though till discharge from the hospital and during follow-up visits. Assistance was provided in preparing the essential documents/paperwork/contacts for the funding agencies including both governmental (Chief-Minister/Prime-Minister/President) and NGO sources. In addition, a detailed questionnaire was filled up for economic assessment of direct/opportunity costs during the entire treatment and 12 months follow up period which forms a part of the study called HEPTROC (Health economic evaluation of primary treatment for ovarian cancer) developed in collaboration with economics departments of Universities. Results: In 2015, 102 patients were operated for OC; only 16 patients (15.68 %) had availed funding of a total sum of INR 1640000 through the hospital system for social assessment. Following challenges were faced by majority of the relatives: 1. Gathering important documents/proper contact details for governmental funding bodies and difficulty in following up the current status 3. Late arrival of funds. In contrast in 2016, 104 OC patients underwent surgery; the direct cost of treatment was significantly higher (median, INR 300000- 400000) compared to other GCs (n=274). 98/104 (94.23%) OC patients could be helped to apply for funds and 90/104(86.56%) patients received funding amounting to a total of INR 10897000. There has been a tenfold increase in funds mobilized in 2016 after the introduction of dedicated MSW in GC. So far, in 2017 (till June), 46/54(85.18%) OC patients applied for funds and 37/54(68.51%) patients have received funding. In a qualitative survey, all patients appreciated the role of the MSW who subsequently became the key worker for patient follow up and the chief portal for patient reported outcome monitoring. Data collection quality for the HEPTROC study was improved when questionnaires were administered by the MSW compared to researchers. Conclusion: Introduction of cancer specific MSW can expedite the availability of funds required for cancer patients and it can positively impact on patient satisfaction and outcome reporting. The economic assessment will influence fund allocation and decision for policymaking in ovarian cancer. Acknowledgement: Jivdaya Foundation Dallas, Texas.

Keywords: economic evaluation, funding, medical social worker, ovarian cancer

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Authors: Daniele Lerede, Gianvito Colucci, Matteo Nicoli, Laura Savoldi

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The European Green Deal is the first internationally agreed set of measures to contrast climate change and environmental degradation. Besides the main target of reducing emissions by at least 55% by 2030, it sets the target of accompanying European countries through an energy transition to make the European Union into a modern, resource-efficient, and competitive net-zero emissions economy by 2050, decoupling growth from the use of resources and ensuring a fair adaptation of all social categories to the transformation process. While the general purpose to allow the realization of the purposes of the Green Deal already dates back to 2019, strategies and policies keep being developed coping with recent circumstances and achievements. However, general long-term measures like the Circular Economy Action Plan, the proposals to shift from fossil natural gas to renewable and low-carbon gases, in particular biomethane and hydrogen, and to end the sale of gasoline and diesel cars by 2035, will all have significant effects on energy supply and demand evolution across the next decades. The interactions between energy supply and demand over long-term time frames are usually assessed via energy system models to derive useful insights for policymaking and to address technological choices and research and development. TEMOA-Europe is a newly developed energy system optimization model instance based on the minimization of the total cost of the system under analysis, adopting a technologically integrated, detailed, and explicit formulation and considering the evolution of the system in partial equilibrium in competitive markets with perfect foresight. TEMOA-Europe is developed on the TEMOA platform, an open-source modeling framework totally implemented in Python, therefore ensuring third-party verification even on large and complex models. TEMOA-Europe is based on a single-region representation of the European Union and EFTA countries on a time scale between 2005 and 2100, relying on a set of assumptions for socio-economic developments based on projections by the International Energy Outlook and a large technological dataset including 7 sectors: the upstream and power sectors for the production of all energy commodities and the end-use sectors, including industry, transport, residential, commercial and agriculture. TEMOA-Europe also includes an updated hydrogen module considering its production, storage, transportation, and utilization. Besides, it can rely on a wide set of innovative technologies, ranging from nuclear fusion and electricity plants equipped with CCS in the power sector to electrolysis-based steel production processes and steel in the industrial sector – with a techno-economic characterization based on public literature – to produce insightful energy scenarios and especially to cope with the very long analyzed time scale. The aim of this work is to examine in detail the scheme of measures and policies for the realization of the purposes of the Green Deal and to transform them into a set of constraints and new socio-economic development pathways. Based on them, TEMOA-Europe will be used to produce and comparatively analyze scenarios to assess the consequences of Green Deal-related measures on the future evolution of the energy mix over the whole energy system in an economic optimization environment.

Keywords: European Green Deal, energy system optimization modeling, scenario analysis, TEMOA-Europe

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Authors: Gousilin Leandra Rocha Da Silva, Stéfani T. A. Dantas, Bruna F. Rossi, Erika R. Bonsaglia, Ivana G. Castilho, Terue Sadatsune, Ary Fernandes Júnior, Vera l. M. Rall

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Kidney failure causes decreased diuresis and accumulation of nitrogenous substances in the body. To increase patient survival, hemodialysis is used as a partial substitute for renal function. However, contamination of the water used in this treatment, causing bacteremia in patients, is a worldwide concern. The Burkholderia cepacia complex (Bcc), a group of bacteria with more than 20 species, is frequently isolated from hemodialysis water samples and comprises opportunistic bacteria, affecting immunosuppressed patients, due to its wide variety of virulence factors, in addition to innate resistance to several antimicrobial agents, contributing to the permanence in the hospital environment and to the pathogenesis in the host. The objective of the present work was to characterize molecularly and phenotypically Bcc isolates collected from the water and dialysate of the Hemodialysis Unit and from the blood of patients at a Public Hospital in Botucatu, São Paulo, Brazil, between 2019 and 2021. We used 33 Bcc isolates, previously obtained from blood cultures from patients with bacteremia undergoing hemodialysis treatment (2019-2021) and 24 isolates obtained from water and dialysate samples in a Hemodialysis Unit (same period). The recA gene was sequenced to identify the specific species among the Bcc group. All isolates were tested for the presence of some genes that encode virulence factors such as cblA, esmR, zmpA and zmpB. Considering the epidemiology of the outbreak, the Bcc isolates were molecularly characterized by Multi Locus Sequence Type (MLST) and by pulsed-field gel electrophoresis (PFGE). The verification and quantification of biofilm in a polystyrene microplate were performed by submitting the isolates to different incubation temperatures (20°C, average water temperature and 35°C, optimal temperature for group growth). The antibiogram was performed with disc diffusion tests on agar, using discs impregnated with cefepime (30µg), ceftazidime (30µg), ciprofloxacin (5µg), gentamicin (10µg), imipenem (10µg), amikacin 30µg), sulfametazol/trimethoprim (23.75/1.25µg) and ampicillin/sulbactam (10/10µg). The presence of ZmpB was identified in all isolates, while ZmpA was observed in 96.5% of the isolates, while none of them presented the cblA and esmR genes. The antibiogram of the 33 human isolates indicated that all were resistant to gentamicin, colistin, ampicillin/sulbactam and imipenem. 16 (48.5%) isolates were resistant to amikacin and lower rates of resistance were observed for meropenem, ceftazidime, cefepime, ciprofloxacin and piperacycline/tazobactam (6.1%). All isolates were sensitive to sulfametazol/trimethoprim, levofloxacin and tigecycline. As for the water isolates, resistance was observed only to gentamicin (34.8%) and imipenem (17.4%). According to PFGE results, all isolates obtained from humans and water belonged to the same pulsotype (1), which was identified by recA sequencing as B. cepacia¸, belonging to sequence type ST-767. By observing a single pulse type over three years, one can observe the persistence of this isolate in the pipeline, contaminating patients undergoing hemodialysis, despite the routine disinfection of water with peracetic acid. This persistence is probably due to the production of biofilm, which protects bacteria from disinfectants and, making this scenario more critical, several isolates proved to be multidrug-resistant (resistance to at least three groups of antimicrobials), turning the patient care even more difficult.

Keywords: hemodialysis, burkholderia cepacia, PFGE, MLST, multi drug resistance

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Authors: Sophio Kobauri, Temur Kantaria, Nina Kulikova, David Tugushi, Ramaz Katsarava

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Polymeric nanoparticles-based drug delivery systems and therapeutics have a great potential in the treatment of a numerous diseases, due to they are characterizing the flexible properties which is giving possibility to modify their structures with a complex definition over their structures, compositions and properties. Important characteristics of the polymeric nanoparticles (PNPs) used as drug carriers are high particle’s stability, high carrier capacity, feasibility of encapsulation of both hydrophilic and hydrophobic drugs, and feasibility of variable routes of administration, including oral application and inhalation; NPs are especially effective for intracellular drug delivery since they penetrate into the cells’ interior though endocytosis. A variety of PNPs based drug delivery systems including charged and neutral, degradable and non-degradable polymers of both natural and synthetic origin have been developed. Among these huge varieties the biodegradable PNPs which can be cleared from the body after the fulfillment of their function could be considered as one of the most promising. For intracellular uptake it is highly desirable to have positively charged PNPs since they can penetrate deep into cell membranes. For long-lasting circulation of PNPs in the body it is important they have so called “stealth coatings” to protect them from the attack of immune system of the organism. One of the effective ways to render the PNPs “invisible” for immune system is their PEGylation which represent the process of pretreatment of polyethylene glycol (PEG) on the surface of PNPs. The present work deals with constructing PNPs from amino acid based biodegradable polymers – regular poly(ester amide) (PEA) composed of sebacic acid, leucine and 1,6-hexandiol (labeled as 8L6), cationic PEA composed of sebacic acid, arginine and 1,6-hexandiol (labeled as 8R6), and comb-like co-PEA composed of sebacic acid, malic acid, leucine and 1,6-hexandiol (labeled as PEG-PEA). The PNPs were fabricated using the polymer deposition/solvent displacement (nanoprecipitation) method. The regular PEA 8L6 form stable negatively charged (zeta-potential within 2-12 mV) PNPs of desired size (within 150-200 nm) in the presence of various surfactants (Tween 20, Tween 80, Brij 010, etc.). Blending the PEAs 8L6 and 8R6 gave the 130-140 nm sized positively charged PNPs having zeta-potential within +20 ÷ +28 mV depending 8L6/8R6 ratio. The PEGylating PEA PEG-PEA was synthesized by interaction of epoxy-co-PEA [8L6]0,5-[tES-L6]0,5 with mPEG-amine-2000 The stable and positively charged PNPs were fabricated using pure PEG-PEA as a surfactant. A firm anchoring of the PEG-PEA with 8L6/8R6 based PNPs (owing to a high afinity of the backbones of all three PEAs) provided good stabilization of the NPs. In vitro biocompatibility study of the new PNPs with four different stable cell lines: A549 (human), U-937 (human), RAW264.7 (murine), Hepa 1-6 (murine) showed they are biocompatible. Considering high stability and cell compatibility of the elaborated PNPs one can conclude that they are promising for subsequent therapeutic applications. This work was supported by the joint grant from the Science and Technology Center in Ukraine and Shota Rustaveli National Science Foundation of Georgia #6298 “New biodegradable cationic polymers composed of arginine and spermine-versatile biomaterials for various biomedical applications”.

Keywords: biodegradable poly(ester amide)s, cationic poly(ester amide), pegylating poly(ester amide), nanoparticles

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Authors: Maria Areias, Diogo Simões, Ana Figueiredo, Anishur Rahman, Filipa Figueiredo, João Nunes

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It is expected that close to 7 billion people will live in urban areas by 2050. In order to improve the sustainability of the territories and its transition towards circular economy, it’s necessary to understand its metabolism and promote and guide the entrepreneurship answer. The study of a macroeconomic metabolism involves the quantification of the inputs, outputs and storage of energy, water, materials and wastes for an urban region. This quantification and analysis representing one opportunity for the promotion of green entrepreneurship. There are several methods to assess the environmental impacts of an urban territory, such as human and environmental risk assessment (HERA), life cycle assessment (LCA), ecological footprint assessment (EF), material flow analysis (MFA), physical input-output table (PIOT), ecological network analysis (ENA), multicriteria decision analysis (MCDA) among others. However, no consensus exists about which of those assessment methods are best to analyze the sustainability of these complex systems. Taking into account the weaknesses and needs identified, the CiiM - Circular Innovation Inter-Municipality project aims to define an uniform and globally accepted methodology through the integration of various methodologies and dynamic approaches to increase the efficiency of macroeconomic metabolisms and promoting entrepreneurship in a circular economy. The pilot territory considered in CiiM project has a total area of 969,428 ha, comprising a total of 897,256 inhabitants (about 41% of the population of the Center Region). The main economic activities in the pilot territory, which contribute to a gross domestic product of 14.4 billion euros, are: social support activities for the elderly; construction of buildings; road transport of goods, retailing in supermarkets and hypermarkets; mass production of other garments; inpatient health facilities; and the manufacture of other components and accessories for motor vehicles. The region's business network is mostly constituted of micro and small companies (similar to the Central Region of Portugal), with a total of 53,708 companies identified in the CIM Region of Coimbra (39 large companies), 28,146 in the CIM Viseu Dão Lafões (22 large companies) and 24,953 in CIM Beiras and Serra da Estrela (13 large companies). For the construction of the database was taking into account data available at the National Institute of Statistics (INE), General Directorate of Energy and Geology (DGEG), Eurostat, Pordata, Strategy and Planning Office (GEP), Portuguese Environment Agency (APA), Commission for Coordination and Regional Development (CCDR) and Inter-municipal Community (CIM), as well as dedicated databases. In addition to the collection of statistical data, it was necessary to identify and characterize the different stakeholder groups in the pilot territory that are relevant to the different metabolism components under analysis. The CIIM project also adds the potential of a Geographic Information System (GIS) so that it is be possible to obtain geospatial results of the territorial metabolisms (rural and urban) of the pilot region. This platform will be a powerful visualization tool of flows of products/services that occur within the region and will support the stakeholders, improving their circular performance and identifying new business ideas and symbiotic partnerships.

Keywords: circular economy tools, life cycle assessment macroeconomic metabolism, multicriteria decision analysis, decision support tools, circular entrepreneurship, industrial and regional symbiosis

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Authors: Jatziri Y. Moreno-Martínez, Arturo Galván, Israel Enrique Herrera Díaz, José Ramón Gasca Tirado

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In recent years, for the elevated viaducts in Mexico City, a construction system based on precast/pre-stressed concrete elements has been used, in which the bridge girders are divided in two parts by imposing a hinged support in sections where the bending moments that are originated by the gravity loads in a continuous beam are minimal. Precast concrete girders with dapped ends are a representative sample of a behavior that has complex configurations of stresses that make them more vulnerable to cracking due to flexure–shear interaction. The design procedures for ends of the dapped girders are well established and are based primarily on experimental tests performed for different configurations of reinforcement. The critical failure modes that can govern the design have been identified, and for each of them, the methods for computing the reinforcing steel that is needed to achieve adequate safety against failure have been proposed. Nevertheless, the design recommendations do not include procedures for controlling diagonal cracking at the entrant corner under service loading. These cracks could cause water penetration and degradation because of the corrosion of the steel reinforcement. The lack of visual access to the area makes it difficult to detect this damage and take timely corrective actions. Three-dimensional non-linear numerical models based on Finite Element Method to study the cracking at the entrant corner of dapped-end beams were performed using the software package ANSYS v. 11.0. The cracking was numerically simulated by using the smeared crack approach. The concrete structure was modeled using three-dimensional solid elements SOLID65 capable of cracking in tension and crushing in compression. Drucker-Prager yield surface was used to include the plastic deformations. The longitudinal post-tension was modeled using LINK8 elements with multilinear isotropic hardening behavior using von Misses plasticity. The reinforcement was introduced with smeared approach. The numerical models were calibrated using experimental tests carried out in “Instituto de Ingeniería, Universidad Nacional Autónoma de México”. In these numerical models the characteristics of the specimens were considered: typical solution based on vertical stirrups (hangers) and on vertical and horizontal hoops with a post-tensioned steel which contributed to a 74% of the flexural resistance. The post-tension is given by four steel wires with a 5/8’’ (16 mm) diameter. Each wire was tensioned to 147 kN and induced an average compressive stress of 4.90 MPa on the concrete section of the dapped end. The loading protocol consisted on applying symmetrical loading to reach the service load (180 kN). Due to the good correlation between experimental and numerical models some additional numerical models were proposed by considering different percentages of post-tension in order to find out how much it influences in the appearance of the cracking in the reentrant corner of the dapped-end beams. It was concluded that the increasing of percentage of post-tension decreases the displacements and the cracking in the reentrant corner takes longer to appear. The authors acknowledge at “Universidad de Guanajuato, Campus Celaya-Salvatierra” and the financial support of PRODEP-SEP (UGTO-PTC-460) of the Mexican government. The first author acknowledges at “Instituto de Ingeniería, Universidad Nacional Autónoma de México”.

Keywords: concrete dapped-end beams, cracking control, finite element analysis, postension

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Authors: Sherif Keddis, Rafik Mitry, Norbert Schwesinger

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Intelligent networking of devices has rapidly been gaining importance over the past years and with recent advances in the fields of microcontrollers, integrated circuits and wireless communication, low power applications have emerged, enabling this trend even more. Connected devices provide a much larger database thus enabling highly intelligent and accurate systems. Ensuring safe drinking water is one of the fields that require constant monitoring and can benefit from an increased accuracy. Monitoring is mainly achieved either through complex measures, such as collecting samples from the points of use, or through metering systems typically distant to the points of use which deliver less accurate assessments of the quality of water. Constant metering near the points of use is complicated due to their inaccessibility; e.g. buried water pipes, locked spaces, which makes system maintenance extremely difficult and often unviable. The research presented here attempts to overcome this challenge by providing these systems with enough energy through a flow harvester inside the pipe thus eliminating the maintenance requirements in terms of battery replacements or containment of leakage resulting from wiring such systems. The proposed flow harvester exploits the piezoelectric properties of polyvinylidene difluoride (PVDF) films to convert turbulence induced oscillations into electrical energy. It is intended to be used in standard water pipes with diameters between 0.5 and 1 inch. The working principle of the harvester uses a ring shaped bluff body inside the pipe to induce pressure fluctuations. Additionally the bluff body houses electronic components such as storage, circuitry and RF-unit. Placing the piezoelectric films downstream of that bluff body causes their oscillation which generates electrical charge. The PVDF-film is placed as a multilayered wrap fixed to the pipe wall leaving the top part to oscillate freely inside the flow. The warp, which allows for a larger active, consists of two layers of 30µm thick and 12mm wide PVDF layered alternately with two centered 6µm thick and 8mm wide aluminum foil electrodes. The length of the layers depends on the number of windings and is part of the investigation. Sealing the harvester against liquid penetration is achieved by wrapping it in a ring-shaped LDPE-film and welding the open ends. The fabrication of the PVDF-wraps is done by hand. After validating the working principle using a wind tunnel, experiments have been conducted in water, placing the harvester inside a 1 inch pipe at water velocities of 0.74m/s. To find a suitable placement of the wrap inside the pipe, two forms of fixation were compared regarding their power output. Further investigations regarding the number of windings required for efficient transduction were made. Best results were achieved using a wrap with 3 windings of the active layers which delivers a constant power output of 0.53µW at a 2.3MΩ load and an effective voltage of 1.1V. Considering the extremely low power requirements of sensor applications, these initial results are promising. For further investigations and optimization, machine designs are currently being developed to automate the fabrication and decrease tolerance of the prototypes.

Keywords: maintenance-free sensors, measurements at point of use, piezoelectric flow harvesting, universal micro generator, wireless metering systems

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Authors: Sergey Kozlov, Juliya Kozlova

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Introduction: The increased attention of researchers to an explosive trauma around the world is associated with a constant renewal of military weapons and a significant increase in terrorist activities using explosive devices. Explosive wave is a well known damaging factor of explosion. The most sensitive to the action of explosive wave in the human body are the head brain, lungs, intestines, urine bladder. The severity of damage to these organs depends on the distance from the explosion epicenter to the object, the power of the explosion, presence of barriers, parameters of the body position, and the presence of protective clothing. One of the places where a shock wave acts, in human tissues and organs, is the vascular endothelial barrier, which suffers the greatest damage in the head brain and lungs. The objective of the study was to determine the pathomorphological changes of the head brain followed the action of explosive wave. Materials and methods of research: To achieve the purpose of the study, there have been studied 6 male corpses delivered to the morgue of Municipal Institution "Dnipropetrovsk regional forensic bureau" during 2014-2016 years. The cause of death of those killed was a military explosive injury. After a visual external assessment of the head brain, for histological study there was conducted the 1 x 1 x 1 cm/piece sampling from different parts of the head brain, i.e. the frontal, parietal, temporal, occipital sites, and also from the cerebellum, pons, medulla oblongata, thalamus, walls of the lateral ventricles, the bottom of the 4th ventricle. Pieces of the head brain were immersed in 10% formalin solution for 24 hours. After fixing, the paraffin blocks were made from the material using the standard method. Then, using a microtome, there were made sections of 4-6 micron thickness from paraffin blocks which then were stained with hematoxylin and eosin. Microscopic analysis was performed using a light microscope with x4, x10, x40 lenses. Results of the study: According to the results of our study, injuries of the head brain were divided into macroscopic and microscopic. Macroscopic injuries were marked according to the results of visual assessment of haemorrhages under the membranes and into the substance, their nature, and localisation, areas of softening. In the microscopic study, our attention was drawn to both vascular changes and those of neurons and glial cells. Microscopic qualitative analysis of histological sections of different parts of the head brain revealed a number of structural changes both at the cellular and tissue levels. Typical changes in most of the studied areas of the head brain included damages of the vascular system. The most characteristic microscopic sign was the separation of vascular walls from neuroglia with the formation of perivascular space. Along with this sign, wall fragmentation of these vessels, haemolysis of erythrocytes, formation of haemorrhages in the newly formed perivascular spaces were found. In addition to damages of the cerebrovascular system, destruction of the neurons, presence of oedema of the brain tissue were observed in the histological sections of the brain. On some sections, the head brain had a heterogeneous step-like or wave-like nature. Conclusions: The pathomorphological microscopic changes in the brain, identified in the study on the died of explosive traumas, can be used for diagnostic purposes in conjunction with other characteristic signs of explosive trauma in forensic and pathological studies. The complex of microscopic signs in the head brain, i.e. separation of blood vessel walls from neuroglia with the perivascular space formation, fragmentation of walls of these blood vessels, erythrocyte haemolysis, formation of haemorrhages in the newly formed perivascular spaces is the direct indication of explosive wave action.

Keywords: blast wave, neurotrauma, human, brain

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Authors: Mohammadjavad Sotoudeheian, Navid Farahmandian

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Cardiac hypertrophy arises in response to persistent increases in hemodynamic loads. In comparison, diabetic cardiomyopathy is defined by an abnormal myocardial changes without other cardiac-related risk factors. Pathological cardiac hypertrophy and myocardial remodeling are hallmarks of cardiovascular diseases and are risk factors for heart failure. The transcription factor forkhead box class O (FOXOs) can protect heart tissue by hostile oxidative stress and stimulating apoptosis and autophagy. FOXO proteins, as sensitive elements and mediators in response to environmental changes, have been revealed to prevent and inverse cardiac hypertrophy. FOXOs are inhibited by insulin and are critical mediators of insulin action. Insulin deficiency and uncontrolled diabetes lead to a catabolic state. FOXO1 acts downstream of the insulin-dependent pathways, which are dysregulated in diabetes. It regulates cardiomyocyte hypertrophy downstream of IGF1R/PI3K/Akt activation, which are critical regulators of cardiac hypertrophy. The complex network of signaling pathways comprising insulin/IGF-1 signaling, AMPK, JNK, and Sirtuins regulate the development of cardiovascular dysfunction by modulating the activity of FOXOs. Insulin receptors and IGF1R act via the PI3k/Akt and the MAPK/ERK pathways. Activation of Akt in response to insulin or IGF-1 induces phosphorylation of FOXOs. Increased protein synthesis induced by activation of the IGF-I/Akt/mTOR signaling pathway leads to hypertrophy. This pathway and the myostatin/Smad pathway are potent negative muscle development regulators. In cardiac muscle, insulin receptor substrates (IRS)-1 or IRS-2 activates the Akt signaling pathway and inactivate FOXO1. Under metabolic stress, p38 MAPK promotes degradation of IRS-1 and IRS-2 in cardiac myocytes and activates FOXO1, leading to cardiomyopathy. Sirt1 and FOXO1 interaction play an essential role in starvation-induced autophagy in cardiac metabolism. Inhibition of Angiotensin-II induced cardiomyocyte hypertrophy is associated with reduced FOXO1 acetylation and activation of Sirt1. The NF-κB, ERK, and FOXOs are de-acetylated by SIRT1. De-acetylation of FOXO1 induces the expression of genes involved in autophagy and stimulates autophagy flux. Therefore, under metabolic stress, FOXO1 can cause diabetic cardiomyopathy. The overexpression of FOXO1 leads to decreased cardiomyocyte size and suppresses cardiac hypertrophy through inhibition of the calcineurin–NFAT pathway. Diabetes mellitus is associated with elevation of O-GlcNAcylation. Some of its binding partners regulate the substrate selectivity of O-GlcNAc transferase (OGT). O-GlcNAcylation of essential contractile proteins may inhibit protein-protein interactions, reduce calcium sensitivity, and modulate contractile function. Uridine diphosphate (UDP)-GlcNAc is the obligatory substrate of OGT, which catalyzes a reversible post-translational protein modification. The increase of O-GlcNAcylation is accompanied by impaired cardiac hypertrophy in diabetic hearts. Inhibition of O-GlcNAcylation blocks activation of ERK1/2 and hypertrophic growth. O-GlcNAc modification on NFAT is required for its translocation from the cytosol to the nucleus, where NFAT stimulates the transcription of various hypertrophic genes. Inhibition of O-GlcNAcylation dampens NFAT-induced cardiac hypertrophic growth. Transcriptional activity of FOXO1 is enriched by improved O-GlcNAcylation upon high glucose stimulation or OGT overexpression. In diabetic conditions, the modification of FOXO1 by O-GlcNAc is promoted in cardiac troponin I and myosin light chain 2. Therefore targeting O-GlcNAcylation represents a potential therapeutic option to prevent hypertrophy in the diabetic heart.

Keywords: diabetes, cardiac hypertrophy, O-GlcNAcylation, FOXO1, Akt, PI3K, AMPK, insulin

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Authors: Daniel Stabler, Anne-Laure Mention, Henri Hakala, Ahmad Alaassar

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In Europe, 2.2 billion tons of waste annually generate severe environmental damage and economic burdens, and negatively impact human health. A stark illustration of the problem is found within the consumer electronics industry, which reflects one of the most complex global waste streams. Of the 5.3 billion globally discarded mobile phones in 2022, only 17% were properly recycled. To address these pressing issues, Europe has made significant strides in developing waste management strategies, Circular Economy initiatives, and Right to Repair policies. These endeavors aim to make product repair and maintenance more accessible, extend product lifespans, reduce waste, and promote sustainable resource use. European countries have introduced Right to Repair policies, often in conjunction with extended producer responsibility legislation, repair subsidies, and consumer repair indices, to varying degrees of regulatory rigor. Changing societal trends emphasizing sustainability and environmental responsibility have driven consumer demand for more sustainable and repairable products, benefiting repair-focused consumer electronics businesses. In academic research, much of the literature in Management studies has examined the European Circular Economy and the Right to Repair from firm-level perspectives. These studies frequently employ a business-model lens, emphasizing innovation and strategy frameworks. However, this study takes an institutional perspective, aiming to understand the adoption of Circular Economy and repair-focused business models within the European consumer electronics market. The concepts of the Circular Economy and the Right to Repair align with institutionalism as they reflect evolving societal norms favoring sustainability and consumer empowerment. Regulatory institutions play a pivotal role in shaping and enforcing these concepts through legislation, influencing the behavior of businesses and individuals. Compliance and enforcement mechanisms are essential for their success, compelling actors to adopt sustainable practices and consider product life extension. Over time, these mechanisms create a path for more sustainable choices, underscoring the influence of institutions and societal values on behavior and decision-making. Institutionalism, particularly 'neo-institutionalism,' provides valuable insights into the factors driving the adoption of Circular and repair-focused business models. Neo-institutional pressures can manifest through coercive regulatory initiatives or normative standards shaped by socio-cultural trends. The Right to Repair movement has emerged as a prominent and influential idea within academic discourse and sustainable development initiatives. Therefore, understanding how macro-level societal shifts toward the Circular Economy and the Right to Repair trigger firm-level responses is imperative. This study aims to answer a crucial question about the impact of European Right to Repair initiatives had on the financial and environmental performance of European consumer electronics companies at the firm level. A quantitative and statistical research design will be employed. The study will encompass an extensive sample of consumer electronics firms in Northern and Western Europe, analyzing their financial and environmental performance in relation to the implementation of Right to Repair mechanisms. The study's findings are expected to provide valuable insights into the broader implications of the Right to Repair and Circular Economy initiatives on the European consumer electronics industry.

Keywords: circular economy, right to repair, institutionalism, environmental management, european union

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Authors: K. Raghu, M. N. Vathhsala, Naveen Aradya, Sharth

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River sand is the most preferred fine aggregate for mortar and concrete. River sand is a product of natural weathering of rocks over a period of millions of years and is mined from river beds. Sand mining has disastrous environmental consequences. The excessive mining of river bed is creating an ecological imbalance. This has lead to have restrictions imposed by ministry of environment on sand mining. Driven by the acute need for sand, stone dust or manufactured sand prepared from the crushing and screening of coarse aggregate is being used as sand in the recent past. However manufactured sand is also a natural material and has quarrying and quality issues. To reduce the burden on the environment, alternative materials to be used as fine aggregates are being extensively investigated all over the world. Looking to the quantum of requirements, quality and properties there has been a global consensus on a material – Granulated slags. Granulated slag has been proven as a suitable material for replacing natural sand / crushed fine aggregates. In developed countries, the use of granulated slag as fine aggregate to replace natural sand is well established and is in regular practice. In the present paper Granulated slag has been experimented for usage in mortar. Slags are the main by-products generated during iron and steel production in the steel industry. Over the past decades, the steel production has increased and, consequently, the higher volumes of by-products and residues generated which have driven to the reuse of these materials in an increasingly efficient way. In recent years new technologies have been developed to improve the recovery rates of slags. Increase of slags recovery and use in different fields of applications like cement making, construction and fertilizers help in preserving natural resources. In addition to the environment protection, these practices produced economic benefits, by providing sustainable solutions that can allow the steel industry to achieve its ambitious targets of “zero waste” in coming years. Slags are generated at two different stages of steel production, iron making and steel making known as BF(Blast Furnace) slag and steel slag respectively. The slagging agent or fluxes, such as lime stone, dolomite and quartzite added into BF or steel making furnaces in order to remove impurities from ore, scrap and other ferrous charges during smelting. The slag formation is the result of a complex series of physical and chemical reactions between the non-metallic charge(lime stone, dolomite, fluxes), the energy sources(coal, coke, oxygen, etc.) and refractory materials. Because of the high temperatures (about 15000 C) during their generation, slags do not contain any organic substances. Due to the fact that slags are lighter than the liquid metal, they float and get easily removed. The slags protect the metal bath from atmosphere and maintain temperature through a kind of liquid formation. These slags are in liquid state and solidified in air after dumping in the pit or granulated by impinging water systems. Generally, BF slags are granulated and used in cement making due to its high cementious properties, and steel slags are mostly dumped due to unfavourable physio-chemical conditions. The increasing dump of steel slag not only occupies a plenty of land but also wastes resources and can potentially have an impact on the environment due to water pollution. Since BF slag contains little Fe and can be used directly. BF slag has found a wide application, such as cement production, road construction, Civil Engineering work, fertilizer production, landfill daily cover, soil reclamation, prior to its application outside the iron and steel making process.

Keywords: steel slag, river sand, granulated slag, environmental

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Authors: Irina Alexandra Paun, Mona Mihailescu, Marian Zamfirescu, Catalin Romeo Luculescu, Adriana Maria Acasandrei, Cosmin Catalin Mustaciosu, Roxana Cristina Popescu, Maria Dinescu

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A major concern in bone tissue engineering is to develop complex 3D architectures that mimic the natural cells environment, facilitate the cells growth in a defined manner and allow the flow transport of nutrients and metabolic waste. In particular, porous structures of controlled pore size and positioning are indispensable for growing human-like bone structures. Another concern is to monitor both the structures and the seeded cells with high spatial resolution and without interfering with the cells natural environment. The present approach relies on laser-based technologies employed for fabricating porous biomimetic structures that support the growth of osteoblast-like cells and for their non-invasive 3D imaging. Specifically, the porous structures were built by two photon polymerization –direct writing (2PP_DW) of the commercially available photoresists IL-L780, using the Photonic Professional 3D lithography system. The structures consist of vertical tubes with micrometer-sized heights and diameters, in a honeycomb-like spatial arrangement. These were fabricated by irradiating the IP-L780 photoresist with focused laser pulses with wavelength centered at 780 nm, 120 fs pulse duration and 80 MHz repetition rate. The samples were precisely scanned in 3D by piezo stages. The coarse positioning was done by XY motorized stages. The scanning path was programmed through a writing language (GWL) script developed by Nanoscribe. Following laser irradiation, the unexposed regions of the photoresist were washed out by immersing the samples in the Propylene Glycol Monomethyl Ether Acetate (PGMEA). The porous structures were seeded with osteoblast like MG-63 cells and their osteogenic potential was tested in vitro. The cell-seeded structures were analyzed in 3D using the digital holographic microscopy technique (DHM). DHM is a marker free and high spatial resolution imaging tool, where the hologram acquisition is performed non-invasively i.e. without interfering with the cells natural environment. Following hologram recording, a digital algorithm provided a 3D image of the sample, as well as information about its refractive index, which is correlated with the intracellular content. The axial resolution of the images went down to the nanoscale, while the temporal scales ranged from milliseconds up to hours. The hologram did not involve sample scanning and the whole image was available in one frame recorded going over 200μm field of view. The digital holograms processing provided 3D quantitative information on the porous structures and allowed a quantitative analysis of the cellular response in respect to the porous architectures. The cellular shape and dimensions were found to be influenced by the underlying micro relief. Furthermore, the intracellular content gave evidence on the beneficial role of the porous structures in promoting osteoblast differentiation. In all, the proposed laser-based protocol emerges as a promising tool for the fabrication and non-invasive imaging of porous constructs for bone tissue engineering. Acknowledgments: This work was supported by a grant of the Romanian Authority for Scientific Research and Innovation, CNCS-UEFISCDI, project PN-II-RU-TE-2014-4-2534 (contract 97 from 01/10/2015) and by UEFISCDI PN-II-PT-PCCA no. 6/2012. A part of this work was performed in the CETAL laser facility, supported by the National Program PN 16 47 - LAPLAS IV.

Keywords: biomimetic, holography, laser, osteoblast, two photon polymerization

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Authors: B.L. España-Sánchez, E. Luna-Hernández, R.A. Mauricio-Sánchez, M.E. Cruz-Soto, F. Padilla-Vaca, R. Muñoz, L. Granados-López, L.R. Ovalle-Flores, J.L. Menchaca-Arredondo, G. Luna-Bárcenas

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Treatment of burn injured has been considered an important clinical problem due to the fluid control and the presence of microorganisms during the healing process. Conventional treatment includes antiseptic techniques, topical medication and surgical removal of damaged skin, to avoid bacterial growth. In order to accelerate this process, different alternatives for tissue regeneration have been explored, including artificial skin, polymers, hydrogels and hybrid materials. Some requirements consider a nonreactive organic polymer with high biocompatibility and skin adherence, avoiding bacterial infections. Chitin-derivative biopolymer such as chitosan (CS) has been used in skin regeneration following third-degree burns. The biological interest of CS is associated with the improvement of tissue cell stimulation, biocompatibility and antibacterial properties. In particular, antimicrobial properties of CS can be significantly increased when is blended with nanostructured materials. Silver-based nanocomposites have gained attention in medicine due to their high antibacterial properties against pathogens, related to their high surface area/volume ratio at nanomolar concentrations. Silver nanocomposites can be blended or synthesized with chitin-derivative biopolymers in order to obtain a biodegradable/antimicrobial hybrid with improved physic-mechanical properties. In this study, nanocomposites based on chitosan/silver nanoparticles (CS/nAg) were synthesized by the in situ chemical reduction method, improving their antibacterial properties against pathogenic bacteria and enhancing the healing process in thermal burn injuries produced in an animal model. CS/nAg was prepared in solution by the chemical reduction method, using AgNO₃ as precursor. CS was dissolved in acetic acid and mixed with different molar concentrations of AgNO₃: 0.01, 0.025, 0.05 and 0.1 M. Solutions were stirred at 95°C during 20 hours, in order to promote the nAg formation. CS/nAg solutions were placed in Petri dishes and dried, to obtain films. Structural analyses confirm the synthesis of silver nanoparticles (nAg) by means of UV-Vis and TEM, with an average size of 7.5 nm and spherical morphology. FTIR analyses showed the complex formation by the interaction of hydroxyl and amine groups with metallic nanoparticles, and surface chemical analysis (XPS) shows low concentration of Ag⁰/Ag⁺ species. Topography surface analyses by means of AFM shown that hydrated CS form a mesh with an average diameter of 10 µm. Antibacterial activity against S. aureus and P. aeruginosa was improved in all evaluated conditions, such as nAg loading and interaction time. CS/nAg nanocomposites films did not show Ag⁰/Ag⁺ release in saline buffer and rat serum after exposition during 7 days. Healing process was significantly enhanced by the presence of CS/nAg nanocomposites, inducing the production of myofibloblasts, collagen remodelation, blood vessels neoformation and epidermis regeneration after 7 days of injury treatment, by means of histological and immunohistochemistry assays. The present work suggests that hydrated CS/nAg nanocomposites can be formed a mesh, improving the bacterial penetration and the contact with embedded nAg, producing complete growth inhibition after 1.5 hours. Furthermore, CS/nAg nanocomposites improve the cell tissue regeneration in thermal burn injuries induced in rats. Synthesis of antibacterial, non-toxic, and biocompatible nanocomposites can be an important issue in tissue engineering and health care applications.

Keywords: antibacterial, chitosan, healing process, nanocomposites, silver

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Authors: A. Chandra, M. Arthur, L. Mize, A. Pomeroy-Stevens

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Background: Lower and middle-income countries (LMICs) in Asia face rapid urbanization resulting in both economic opportunities (the urban advantage) and emerging health challenges. Urban health risks are magnified in informal settlements and include infectious disease outbreaks, inadequate access to health services, and poor air quality. Over the coming years, urban spaces in Asia will face accelerating public health risks related to migration, climate change, and environmental health. These challenges are complex and require multi-sectoral and multi-stakeholder solutions. The Building Health Cities (BHC) program is funded by the United States Agency for International Development (USAID) to work with smart city initiatives in the Asia region. BHC approaches urban health challenges by addressing policies, planning, and services through a health equity lens, with a particular focus on informal settlements and migrant communities. The program works to develop data-driven decision-making, build inclusivity through stakeholder engagement, and facilitate the uptake of appropriate technology. Methodology: The BHC program has partnered with the smart city initiatives of Indore in India, Makassar in Indonesia, and Da Nang in Vietnam. Implementing partners support municipalities to improve health delivery and equity using two key approaches: political economy analysis and participatory systems mapping. Political economy analyses evaluate barriers to collective action, including corruption, security, accountability, and incentives. Systems mapping evaluates community health challenges using a cross-sectoral approach, analyzing the impact of economic, environmental, transport, security, health system, and built environment factors. The mapping exercise draws on the experience and expertise of a diverse cohort of stakeholders, including government officials, municipal service providers, and civil society organizations. Results: Systems mapping and political economy analyses identified significant barriers for health care in migrant populations. In Makassar, migrants are unable to obtain the necessary card that entitles them to subsidized health services. This finding is being used to engage with municipal governments to mitigate the barriers that limit migrant enrollment in the public social health insurance scheme. In Indore, the project identified poor drainage of storm and wastewater in migrant settlements as a cause of poor health. Unsafe and inadequate infrastructure placed residents of these settlements at risk for both waterborne diseases and injuries. The program also evaluated the capacity of urban primary health centers serving migrant communities, identifying challenges related to their hours of service and shortages of health workers. In Da Nang, the systems mapping process has only recently begun, with the formal partnership launched in December 2019. Conclusion: This paper explores lessons learned from BHC’s systems mapping, political economy analyses, and stakeholder engagement approaches. The paper shares progress related to the health of migrants in informal settlements. Case studies feature barriers identified and mitigating steps, including governance actions, taken by local stakeholders in partner cities. The paper includes an update on ongoing progress from Indore and Makassar and experience from the first six months of program implementation from Da Nang.

Keywords: informal settlements, migration, stakeholder engagement mapping, urban health

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Authors: Savvas Rogotis, Nikolaos Kalatzis, Stergios Dimou-Sakellariou, Nikolaos Marianos

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As precision holds the key for the introduction of distinct benefits in agriculture (e.g., energy savings, reduced labor costs, optimal application of inputs, improved products, and yields), it steadily becomes evident that new initiatives should focus on rendering Precision Agriculture (PA) more accessible to the average farmer. PA leverages on technologies such as the Internet of Things (IoT), earth observation, robotics and positioning systems (e.g., the Global Navigation Satellite System – GNSS - as well as individual positioning systems like GPS, Glonass, Galileo) that allow: from simple data georeferencing to optimal navigation of agricultural machinery to even more complex tasks like Variable Rate Applications. An identified customer pain point is that, from one hand, typical triangulation-based positioning systems are not accurate enough (with errors up to several meters), while on the other hand, high precision positioning systems reaching centimeter-level accuracy, are very costly (up to thousands of euros). Within this paper, a Ground-Based Augmentation System (GBAS) is introduced, that can be adapted to any existing IoT field-sensing station infrastructure. The latter should cover a minimum set of requirements, and in particular, each station should operate as a fixed, obstruction-free towards the sky, energy supplying unit. Station augmentation will allow them to function in pairs with GNSS rovers following the differential GNSS base-rover paradigm. This constitutes a key innovation element for the proposed solution that encompasses differential GNSS capabilities into an IoT field-sensing infrastructure. Integrating this kind of information supports the provision of several additional PA beneficial services such as spatial mapping, route planning, and automatic field navigation of unmanned vehicles (UVs). Right at the heart of the designed system, there is a high-end GNSS toolkit with base-rover variants and Real-Time Kinematic (RTK) capabilities. The GNSS toolkit had to tackle all availability, performance, interfacing, and energy-related challenges that are faced for a real-time, low-power, and reliable in the field operation. Specifically, in terms of performance, preliminary findings exhibit a high rover positioning precision that can even reach less than 10-centimeters. As this precision is propagated to the full dataset collection, it enables tractors, UVs, Android-powered devices, and measuring units to deal with challenging real-world scenarios. The system is validated with the help of Gaiatrons, a mature network of agro-climatic telemetry stations with presence all over Greece and beyond ( > 60.000ha of agricultural land covered) that constitutes part of “gaiasense” (www.gaiasense.gr) smart farming (SF) solution. Gaiatrons constantly monitor atmospheric and soil parameters, thus, providing exact fit to operational requirements asked from modern SF infrastructures. Gaiatrons are ultra-low-cost, compact, and energy-autonomous stations with a modular design that enables the integration of advanced GNSS base station capabilities on top of them. A set of demanding pilot demonstrations has been initiated in Stimagka, Greece, an area with a diverse geomorphological landscape where grape cultivation is particularly popular. Pilot demonstrations are in the course of validating the preliminary system findings in its intended environment, tackle all technical challenges, and effectively highlight the added-value offered by the system in action.

Keywords: GNSS, GBAS, precision agriculture, RTK, smart farming

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