Search results for: Antonio E. de A. Nogueira
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 288

Search results for: Antonio E. de A. Nogueira

48 A Mixed Method Approach for Modeling Entry Capacity at Rotary Intersections

Authors: Antonio Pratelli, Lorenzo Brocchini, Reginald Roy Souleyrette

Abstract:

A rotary is a traffic circle intersection where vehicles entering from branches give priority to circulating flow. Vehicles entering the intersection from converging roads move around the central island and weave out of the circle into their desired exiting branch. This creates merging and diverging conflicts among any entry and its successive exit, i.e., a section. Therefore, rotary capacity models are usually based on the weaving of the different movements in any section of the circle, and the maximum rate of flow value is then related to each weaving section of the rotary. Nevertheless, the single-section capacity value does not lead to the typical performance characteristics of the intersection, such as the entry average delay which is directly linked to its level of service. From another point of view, modern roundabout capacity models are based on the limitation of the flow entering from the single entrance due to the amount of flow circulating in front of the entrance itself. Modern roundabouts capacity models generally lead also to a performance evaluation. This paper aims to incorporate a modern roundabout capacity model into an old rotary capacity method to obtain from the latter the single input capacity and ultimately achieve the related performance indicators. Put simply; the main objective is to calculate the average delay of each single roundabout entrance to apply the most common Highway Capacity Manual, or HCM, criteria. The paper is organized as follows: firstly, the rotary and roundabout capacity models are sketched, and it has made a brief introduction to the model combination technique with some practical instances. The successive section is deserved to summarize the TRRL old rotary capacity model and the most recent HCM-7th modern roundabout capacity model. Then, the two models are combined through an iteration-based algorithm, especially set-up and linked to the concept of roundabout total capacity, i.e., the value reached due to a traffic flow pattern leading to the simultaneous congestion of all roundabout entrances. The solution is the average delay for each entrance of the rotary, by which is estimated its respective level of service. In view of further experimental applications, at this research stage, a collection of existing rotary intersections operating with the priority-to-circle rule has already started, both in the US and in Italy. The rotaries have been selected by direct inspection of aerial photos through a map viewer, namely Google Earth. Each instance has been recorded by location, general urban or rural, and its main geometrical patterns. Finally, conclusion remarks are drawn, and a discussion on some further research developments has opened.

Keywords: mixed methods, old rotary and modern roundabout capacity models, total capacity algorithm, level of service estimation

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47 Nanomechanical Characterization of Healthy and Tumor Lung Tissues at Cell and Extracellular Matrix Level

Authors: Valeria Panzetta, Ida Musella, Sabato Fusco, Paolo Antonio Netti

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The study of the biophysics of living cells drew attention to the pivotal role of the cytoskeleton in many cell functions, such as mechanics, adhesion, proliferation, migration, differentiation and neoplastic transformation. In particular, during the complex process of malignant transformation and invasion cell cytoskeleton devolves from a rigid and organized structure to a more compliant state, which confers to the cancer cells a great ability to migrate and adapt to the extracellular environment. In order to better understand the malignant transformation process from a mechanical point of view, it is necessary to evaluate the direct crosstalk between the cells and their surrounding extracellular matrix (ECM) in a context which is close to in vivo conditions. In this study, human biopsy tissues of lung adenocarcinoma were analyzed in order to define their mechanical phenotype at cell and ECM level, by using particle tracking microrheology (PTM) technique. Polystyrene beads (500 nm) were introduced into the sample slice. The motion of beads was obtained by tracking their displacements across cell cytoskeleton and ECM structures and mean squared displacements (MSDs) were calculated from bead trajectories. It has been already demonstrated that the amplitude of MSD is inversely related to the mechanical properties of intracellular and extracellular microenvironment. For this reason, MSDs of particles introduced in cytoplasm and ECM of healthy and tumor tissues were compared. PTM analyses showed that cancerous transformation compromises mechanical integrity of cells and extracellular matrix. In particular, the MSD amplitudes in cells of adenocarcinoma were greater as compared to cells of normal tissues. The increased motion is probably associated to a less structured cytoskeleton and consequently to an increase of deformability of cells. Further, cancer transformation is also accompanied by extracellular matrix stiffening, as confirmed by the decrease of MSDs of matrix in tumor tissue, a process that promotes tumor proliferation and invasiveness, by activating typical oncogenic signaling pathways. In addition, a clear correlation between MSDs of cells and tumor grade was found. MSDs increase when tumor grade passes from 2 to 3, indicating that cells undergo to a trans-differentiation process during tumor progression. ECM stiffening is not dependent on tumor grade, but the tumor stage resulted to be strictly correlated with both cells and ECM mechanical properties. In fact, a greater stage is assigned to tumor spread to regional lymph nodes and characterized by an up-regulation of different ECM proteins, such as collagen I fibers. These results indicate that PTM can be used to get nanomechanical characterization at different scale levels in an interpretative and diagnostic context.

Keywords: cytoskeleton, extracellular matrix, mechanical properties, particle tracking microrheology, tumor

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46 New Bio-Strategies for Ochratoxin a Detoxification Using Lactic Acid Bacteria

Authors: José Maria, Vânia Laranjo, Luís Abrunhosa, António Inês

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The occurrence of mycotoxigenic moulds such as Aspergillus, Penicillium and Fusarium in food and feed has an important impact on public health, by the appearance of acute and chronic mycotoxicoses in humans and animals, which is more severe in the developing countries due to lack of food security, poverty and malnutrition. This mould contamination also constitutes a major economic problem due the lost of crop production. A great variety of filamentous fungi is able to produce highly toxic secondary metabolites known as mycotoxins. Most of the mycotoxins are carcinogenic, mutagenic, neurotoxic and immunosuppressive, being ochratoxin A (OTA) one of the most important. OTA is toxic to animals and humans, mainly due to its nephrotoxic properties. Several approaches have been developed for decontamination of mycotoxins in foods, such as, prevention of contamination, biodegradation of mycotoxins-containing food and feed with microorganisms or enzymes and inhibition or absorption of mycotoxin content of consumed food into the digestive tract. Some group of Gram-positive bacteria named lactic acid bacteria (LAB) are able to release some molecules that can influence the mould growth, improving the shelf life of many fermented products and reducing health risks due to exposure to mycotoxins. Some LAB are capable of mycotoxin detoxification. Recently our group was the first to describe the ability of LAB strains to biodegrade OTA, more specifically, Pediococcus parvulus strains isolated from Douro wines. The pathway of this biodegradation was identified previously in other microorganisms. OTA can be degraded through the hydrolysis of the amide bond that links the L-β-phenylalanine molecule to the ochratoxin alpha (OTα) a non toxic compound. It is known that some peptidases from different origins can mediate the hydrolysis reaction like, carboxypeptidase A an enzyme from the bovine pancreas, a commercial lipase and several commercial proteases. So, we wanted to have a better understanding of this OTA degradation process when LAB are involved and identify which molecules where present in this process. For achieving our aim we used some bioinformatics tools (BLAST, CLUSTALX2, CLC Sequence Viewer 7, Finch TV). We also designed specific primers and realized gene specific PCR. The template DNA used came from LAB strains samples of our previous work, and other DNA LAB strains isolated from elderberry fruit, silage, milk and sausages. Through the employment of bioinformatics tools it was possible to identify several proteins belonging to the carboxypeptidase family that participate in the process of OTA degradation, such as serine type D-Ala-D-Ala carboxypeptidase and membrane carboxypeptidase. In conclusions, this work has identified carboxypeptidase proteins being one of the molecules present in the OTA degradation process when LAB are involved.

Keywords: carboxypeptidase, lactic acid bacteria, mycotoxins, ochratoxin a.

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45 Experimental Study of Vibration Isolators Made of Expanded Cork Agglomerate

Authors: S. Dias, A. Tadeu, J. Antonio, F. Pedro, C. Serra

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The goal of the present work is to experimentally evaluate the feasibility of using vibration isolators made of expanded cork agglomerate. Even though this material, also known as insulation cork board (ICB), has mainly been studied for thermal and acoustic insulation purposes, it has strong potential for use in vibration isolation. However, the adequate design of expanded cork blocks vibration isolators will depend on several factors, such as excitation frequency, static load conditions and intrinsic dynamic behavior of the material. In this study, transmissibility tests for different static and dynamic loading conditions were performed in order to characterize the material. Since the material’s physical properties can influence the vibro-isolation performance of the blocks (in terms of density and thickness), this study covered four mass density ranges and four block thicknesses. A total of 72 expanded cork agglomerate specimens were tested. The test apparatus comprises a vibration exciter connected to an excitation mass that holds the test specimen. The test specimens under characterization were loaded successively with steel plates in order to obtain results for different masses. An accelerometer was placed at the top of these masses and at the base of the excitation mass. The test was performed for a defined frequency range, and the amplitude registered by the accelerometers was recorded in time domain. For each of the signals (signal 1- vibration of the excitation mass, signal 2- vibration of the loading mass) a fast Fourier transform (FFT) was applied in order to obtain the frequency domain response. For each of the frequency domain signals, the maximum amplitude reached was registered. The ratio between the amplitude (acceleration) of signal 2 and the amplitude of signal 1, allows the calculation of the transmissibility for each frequency. Repeating this procedure allowed us to plot a transmissibility curve for a certain frequency range. A number of transmissibility experiments were performed to assess the influence of changing the mass density and thickness of the expanded cork blocks and the experimental conditions (static load and frequency of excitation). The experimental transmissibility tests performed in this study showed that expanded cork agglomerate blocks are a good option for mitigating vibrations. It was concluded that specimens with lower mass density and larger thickness lead to better performance, with higher vibration isolation and a larger range of isolated frequencies. In conclusion, the study of the performance of expanded cork agglomerate blocks presented herein will allow for a more efficient application of expanded cork vibration isolators. This is particularly relevant since this material is a more sustainable alternative to other commonly used non-environmentally friendly products, such as rubber.

Keywords: expanded cork agglomerate, insulation cork board, transmissibility tests, sustainable materials, vibration isolators

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44 Computational Code for Solving the Navier-Stokes Equations on Unstructured Meshes Applied to the Leading Edge of the Brazilian Hypersonic Scramjet 14-X

Authors: Jayme R. T. Silva, Paulo G. P. Toro, Angelo Passaro, Giannino P. Camillo, Antonio C. Oliveira

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An in-house C++ code has been developed, at the Prof. Henry T. Nagamatsu Laboratory of Aerothermodynamics and Hypersonics from the Institute of Advanced Studies (Brazil), to estimate the aerothermodynamic properties around the Hypersonic Vehicle Integrated to the Scramjet. In the future, this code will be applied to the design of the Brazilian Scramjet Technological Demonstrator 14-X B. The first step towards accomplishing this objective, is to apply the in-house C++ code at the leading edge of a flat plate, simulating the leading edge of the 14-X Hypersonic Vehicle, making possible the wave phenomena of oblique shock and boundary layer to be analyzed. The development of modern hypersonic space vehicles requires knowledge regarding the characteristics of hypersonic flows in the vicinity of a leading edge of lifting surfaces. The strong interaction between a shock wave and a boundary layer, in a high supersonic Mach number 4 viscous flow, close to the leading edge of the plate, considering no slip condition, is numerically investigated. The small slip region is neglecting. The study consists of solving the fluid flow equations for unstructured meshes applying the SIMPLE algorithm for Finite Volume Method. Unstructured meshes are generated by the in-house software ‘Modeler’ that was developed at Virtual’s Engineering Laboratory from the Institute of Advanced Studies, initially developed for Finite Element problems and, in this work, adapted to the resolution of the Navier-Stokes equations based on the SIMPLE pressure-correction scheme for all-speed flows, Finite Volume Method based. The in-house C++ code is based on the two-dimensional Navier-Stokes equations considering non-steady flow, with nobody forces, no volumetric heating, and no mass diffusion. Air is considered as calorically perfect gas, with constant Prandtl number and Sutherland's law for the viscosity. Solutions of the flat plate problem for Mach number 4 include pressure, temperature, density and velocity profiles as well as 2-D contours. Also, the boundary layer thickness, boundary conditions, and mesh configurations are presented. The same problem has been solved by the academic license of the software Ansys Fluent and for another C++ in-house code, which solves the fluid flow equations in structured meshes, applying the MacCormack method for Finite Difference Method, and the results will be compared.

Keywords: boundary-layer, scramjet, simple algorithm, shock wave

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43 Study of the Kinetics of Formation of Carboxylic Acids Using Ion Chromatography during Oxidation Induced by Rancimat of the Oleic Acid, Linoleic Acid, Linolenic Acid, and Biodiesel

Authors: Patrícia T. Souza, Marina Ansolin, Eduardo A. C. Batista, Antonio J. A. Meirelles, Matthieu Tubino

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Lipid oxidation is a major cause of the deterioration of the quality of the biodiesel, because the waste generated damages the engines. Among the main undesirable effects are the increase of viscosity and acidity, leading to the formation of insoluble gums and sediments which cause the blockage of fuel filters. The auto-oxidation is defined as the spontaneous reaction of atmospheric oxygen with lipids. Unsaturated fatty acids are usually the components affected by such reactions. They are present as free fatty acids, fatty esters and glycerides. To determine the oxidative stability of biodiesels, through the induction period, IP, the Rancimat method is used, which allows continuous monitoring of the induced oxidation process of the samples. During the oxidation of the lipids, volatile organic acids are produced as byproducts, in addition, other byproducts, including alcohols and carbonyl compounds, may be further oxidized to carboxylic acids. By the methodology developed in this work using ion chromatography, IC, analyzing the water contained in the conductimetric vessel, were quantified organic anions of carboxylic acids in samples subjected to oxidation induced by Rancimat. The optimized chromatographic conditions were: eluent water:acetone (80:20 v/v) with 0.5 mM sulfuric acid; flow rate 0.4 mL min-1; injection volume 20 µL; eluent suppressor 20 mM LiCl; analytical curve from 1 to 400 ppm. The samples studied were methyl biodiesel from soybean oil and unsaturated fatty acids standards: oleic, linoleic and linolenic. The induced oxidation kinetics curves were constructed by analyzing the water contained in the conductimetric vessels which were removed, each one, from the Rancimat apparatus at prefixed intervals of time. About 3 g of sample were used under the conditions of 110 °C and air flow rate of 10 L h-1. The water of each conductimetric Rancimat measuring vessel, where the volatile compounds were collected, was filtered through a 0.45 µm filter and analyzed by IC. Through the kinetic data of the formation of the organic anions of carboxylic acids, the formation rates of the same were calculated. The observed order of the rates of formation of the anions was: formate >>> acetate > hexanoate > valerate for the oleic acid; formate > hexanoate > acetate > valerate for the linoleic acid; formate >>> valerate > acetate > propionate > butyrate for the linolenic acid. It is possible to suppose that propionate and butyrate are obtained mainly from linolenic acid and that hexanoate is originated from oleic and linoleic acid. For the methyl biodiesel the order of formation of anions was: formate >>> acetate > valerate > hexanoate > propionate. According to the total rate of formation these anions produced during the induced degradation of the fatty acids can be assigned the order of reactivity: linolenic acid > linoleic acid >>> oleic acid.

Keywords: anions of carboxylic acids, biodiesel, ion chromatography, oxidation

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42 Time-Domain Nuclear Magnetic Resonance as a Potential Analytical Tool to Assess Thermisation in Ewe's Milk

Authors: Alessandra Pardu, Elena Curti, Marco Caredda, Alessio Dedola, Margherita Addis, Massimo Pes, Antonio Pirisi, Tonina Roggio, Sergio Uzzau, Roberto Anedda

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Some of the artisanal cheeses products of European Countries certificated as PDO (Protected Designation of Origin) are made from raw milk. To recognise potential frauds (e.g. pasteurisation or thermisation of milk aimed at raw milk cheese production), the alkaline phosphatase (ALP) assay is currently applied only for pasteurisation, although it is known to have notable limitations for the validation of ALP enzymatic state in nonbovine milk. It is known that frauds considerably impact on customers and certificating institutions, sometimes resulting in a damage of the product image and potential economic losses for cheesemaking producers. Robust, validated, and univocal analytical methods are therefore needed to allow Food Control and Security Organisms, to recognise a potential fraud. In an attempt to develop a new reliable method to overcome this issue, Time-Domain Nuclear Magnetic Resonance (TD-NMR) spectroscopy has been applied in the described work. Daily fresh milk was analysed raw (680.00 µL in each 10-mm NMR glass tube) at least in triplicate. Thermally treated samples were also produced, by putting each NMR tube of fresh raw milk in water pre-heated at temperatures from 68°C up to 72°C and for up to 3 min, with continuous agitation, and quench-cooled to 25°C in a water and ice solution. Raw and thermally treated samples were analysed in terms of 1H T2 transverse relaxation times with a CPMG sequence (Recycle Delay: 6 s, interpulse spacing: 0.05 ms, 8000 data points) and quasi-continuous distributions of T2 relaxation times were obtained by CONTIN analysis. In line with previous data collected by high field NMR techniques, a decrease in the spin-spin relaxation constant T2 of the predominant 1H population was detected in heat-treated milk as compared to raw milk. The decrease of T2 parameter is consistent with changes in chemical exchange and diffusive phenomena, likely associated to changes in milk protein (i.e. whey proteins and casein) arrangement promoted by heat treatment. Furthermore, experimental data suggest that molecular alterations are strictly dependent on the specific heat treatment conditions (temperature/time). Such molecular variations in milk, which are likely transferred to cheese during cheesemaking, highlight the possibility to extend the TD-NMR technique directly on cheese to develop a method for assessing a fraud related to the use of a milk thermal treatment in PDO raw milk cheese. Results suggest that TDNMR assays might pave a new way to the detailed characterisation of heat treatments of milk.

Keywords: cheese fraud, milk, pasteurisation, TD-NMR

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41 Optimization of Ultrasound-Assisted Extraction of Oil from Spent Coffee Grounds Using a Central Composite Rotatable Design

Authors: Malek Miladi, Miguel Vegara, Maria Perez-Infantes, Khaled Mohamed Ramadan, Antonio Ruiz-Canales, Damaris Nunez-Gomez

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Coffee is the second consumed commodity worldwide, yet it also generates colossal waste. Proper management of coffee waste is proposed by converting them into products with higher added value to achieve sustainability of the economic and ecological footprint and protect the environment. Based on this, a study looking at the recovery of coffee waste is becoming more relevant in recent decades. Spent coffee grounds (SCG's) resulted from brewing coffee represents the major waste produced among all coffee industry. The fact that SCGs has no economic value be abundant in nature and industry, do not compete with agriculture and especially its high oil content (between 7-15% from its total dry matter weight depending on the coffee varieties, Arabica or Robusta), encourages its use as a sustainable feedstock for bio-oil production. The bio-oil extraction is a crucial step towards biodiesel production by the transesterification process. However, conventional methods used for oil extraction are not recommended due to their high consumption of energy, time, and generation of toxic volatile organic solvents. Thus, finding a sustainable, economical, and efficient extraction technique is crucial to scale up the process and to ensure more environment-friendly production. Under this perspective, the aim of this work was the statistical study to know an efficient strategy for oil extraction by n-hexane using indirect sonication. The coffee waste mixed Arabica and Robusta, which was used in this work. The temperature effect, sonication time, and solvent-to-solid ratio on the oil yield were statistically investigated as dependent variables by Central Composite Rotatable Design (CCRD) 23. The results were analyzed using STATISTICA 7 StatSoft software. The CCRD showed the significance of all the variables tested (P < 0.05) on the process output. The validation of the model by analysis of variance (ANOVA) showed good adjustment for the results obtained for a 95% confidence interval, and also, the predicted values graph vs. experimental values confirmed the satisfactory correlation between the model results. Besides, the identification of the optimum experimental conditions was based on the study of the surface response graphs (2-D and 3-D) and the critical statistical values. Based on the CCDR results, 29 ºC, 56.6 min, and solvent-to-solid ratio 16 were the better experimental conditions defined statistically for coffee waste oil extraction using n-hexane as solvent. In these conditions, the oil yield was >9% in all cases. The results confirmed the efficiency of using an ultrasound bath in extracting oil as a more economical, green, and efficient way when compared to the Soxhlet method.

Keywords: coffee waste, optimization, oil yield, statistical planning

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40 Recent Advances in Research on Carotenoids: From Agrofood Production to Health Outcomes

Authors: Antonio J. Melendez-Martinez

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Beyond their role as natural colorants, some carotenoids are provitamins A and may be involved in health-promoting biological actions and contribute to reducing the risk of developing non-communicable diseases, including several types of cancer, cardiovascular disease, eye conditions, skin disorders or metabolic disorders. Given the versatility of carotenoids, the COST-funded European network to advance carotenoid research and applications in agro-food and health (EUROCAROTEN) is aimed at promoting health through the diet and increasing well-being by means. Stakeholders from 38 countries participate in this network, and one of its main objectives is to promote research on little-studied carotenoids. In this contribution, recent advances of our research group and collaborators in the study of two such understudied carotenoids, namely phytoene and phytofluene, the colorless carotenoids, are outlined. The study of these carotenoids is important as they have been largely neglected despite they are present in our diets, fluids, and tissues, and evidence is accumulating that they may be involved in health-promoting actions. More specifically, studies on their levels in diverse tomato and orange varieties were carried out as well as on their potential bioavailability from different dietary sources. Furthermore, the potential effect of these carotenoids on an animal model subjected to oxidative stress was evaluated. The tomatoes were grown in research greenhouses, and some of them were subjected to regulated deficit irrigation, a sustainable agronomic practice. The citrus samples were obtained from an experimental field. The levels of carotenoids were assessed using HPLC according to routine methodologies followed in our lab. Regarding the potential bioavailability (bioaccessibility) studies, different products containing colorless carotenoids, like fruits, juices, were subjected to simulated in vitro digestions, and their incorporation into mixed micelles was assessed. The effect of the carotenoids on oxidative stress was evaluated on the Caenorhabditis elegans model. For that purpose, the worms were subjected to oxidative stress by means of a hydrogen peroxide challenge. In relation to the presence of colorless carotenoids in tomatoes and orange varieties, it was observed that they are widespread in such products and that there are mutants with very high quantities of them, for instance, the Cara Cara or Pinalate mutant oranges. The studies on their bioaccessibility revealed that, in general, phytoene and phytofluene are more bioaccessible than other common dietary carotenoids, probably due to their distinctive chemical structure. About the in vivo antioxidant capacity of phytoene and phytofluene, it was observed that they both exerted antioxidant effects at certain doses. In conclusion, evidence on the importance of phytoene and phytofluene as dietary easily bioavailable and antioxidant carotenoids has been obtained in recent studies from our group, which can be important shortly to innovate in health-promotion through the development of functional foods and related products.

Keywords: carotenoids, health, functional foods, nutrition, phytoene, phytofluene

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39 Biocompatibility assessment of different origin Barrier Membranes for Guided Bone Regeneration

Authors: Antonio Munar-Frau, Sascha Klismoch, Manfred Schmolz, Federico Hernandez-Alfaro, Jordi Caballe-Serrano

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Introduction: Biocompatibility of biomaterials has been proposed as one of the main criteria for treatment success. For guided bone regeneration (GBR), barrier membranes present a conflict given the number of origins and modifications of these materials. The biologic response to biomaterials is orchestrated by a series of events leading to the integration or rejection of the biomaterial, posing questions such as if a longer occlusive property may trigger an inflammatory reaction. Whole blood cultures are a solution to study the immune response to drugs or biomaterials during the first 24-48 hours. The aim of this study is to determine the early immune response of different origins and chemical modifications of barrier membranes. Materials & Methods: 5 different widely used barrier membranes were included in this study: Acellular dermal matrix (AlloDerm, LifeCell®), Porcine Peritoneum (BioGide, Geistlich Pharma®), Porcine Pericardium (Jason, Botiss Biomaterials GmbH®), Porcine Cross-linked collagen (Ossix Plus, Datum Dental®) and d-PTFE (Cytoplast TXT, Osteogenics Biomedical®). Blood samples were extracted from 3 different healthy donors and incubated with the different samples of barrier membranes for 24 hours. After the incubation time, serum samples were obtained and analyzed by means of biocompatibility assays taking into account 42 markers. Results: In an early stage of the inflammatory response, the Acellular dermal matrix, porcine peritoneum and porcine cross-linked collagen expressed similar patterns of cytokine expression with a great manifestation of ENA 78. Porcine pericardium and d-PTFE presented similar cytokine activation, especially for MMP-3 and MMP-9, although other cytokines were highlighted with lower expression. For the later immune response, Porcine peritoneum and acellular dermal matrix MCP-1 and IL-15 were evident. Porcine pericardium, porcine cross-linked collagen and d-PTFE presented a high expression of IL-16 and lower manifestation of other cytokines. Different behaviors depending on an earlier or later stage of the inflammation process were observed. Barrier membrane inflammatory expression does not only differ depending on the origin, variables such as treatment of the collagen and polymers may also have a great impact on the cytokine expression of the studied barrier membranes during inflammation. Conclusions: Surface treatment and modifications might affect the biocompatibility of the membranes, as different cytokine expressions were evidently depending on the origin of the biomaterial. This study is only a brushstroke regarding the biocompatibility of materials, as it is one of the pioneer studies for ex vivo barrier membranes assays. Studies regarding surface modification are needed in order to clarify mystifications of barrier membrane science.

Keywords: biomaterials, bone regeneration, biocompatibility, inflammation

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38 Integration of a Protective Film to Enhance the Longevity and Performance of Miniaturized Ion Sensors

Authors: Antonio Ruiz Gonzalez, Kwang-Leong Choy

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The measurement of electrolytes has a high value in the clinical routine. Ions are present in all body fluids with variable concentrations and are involved in multiple pathologies such as heart failures and chronic kidney disease. In the case of dissolved potassium, although a high concentration in the blood (hyperkalemia) is relatively uncommon in the general population, it is one of the most frequent acute electrolyte abnormalities. In recent years, the integration of thin films technologies in this field has allowed the development of highly sensitive biosensors with ultra-low limits of detection for the assessment of metals in liquid samples. However, despite the current efforts in the miniaturization of sensitive devices and their integration into portable systems, only a limited number of successful examples used commercially can be found. This fact can be attributed to a high cost involved in their production and the sustained degradation of the electrodes over time, which causes a signal drift in the measurements. Thus, there is an unmet necessity for the development of low-cost and robust sensors for the real-time monitoring of analyte concentrations in patients to allow the early detection and diagnosis of diseases. This paper reports a thin film ion-selective sensor for the evaluation of potassium ions in aqueous samples. As an alternative for this fabrication method, aerosol assisted chemical vapor deposition (AACVD), was applied due to cost-effectivity and fine control over the film deposition. Such a technique does not require vacuum and is suitable for the coating of large surface areas and structures with complex geometries. This approach allowed the fabrication of highly homogeneous surfaces with well-defined microstructures onto 50 nm thin gold layers. The degradative processes of the ubiquitously employed poly (vinyl chloride) membranes in contact with an electrolyte solution were studied, including the polymer leaching process, mechanical desorption of nanoparticles and chemical degradation over time. Rational design of a protective coating based on an organosilicon material in combination with cellulose to improve the long-term stability of the sensors was then carried out, showing an improvement in the performance after 5 weeks. The antifouling properties of such coating were assessed using a cutting-edge quartz microbalance sensor, allowing the quantification of the adsorbed proteins in the nanogram range. A correlation between the microstructural properties of the films with the surface energy and biomolecules adhesion was then found and used to optimize the protective film.

Keywords: hyperkalemia, drift, AACVD, organosilicon

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37 The Participation of Experts in the Criminal Policy on Drugs: The Proposal of a Cannabis Regulation Model in Spain by the Cannabis Policy Studies Group

Authors: Antonio Martín-Pardo

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With regard to the context in which this paper is inserted, it is noteworthy that the current criminal policy model in which we find immersed, denominated by some doctrine sector as the citizen security model, is characterized by a marked tendency towards the discredit of expert knowledge. This type of technic knowledge has been displaced by the common sense and by the daily experience of the people at the time of legislative drafting, as well as by excessive attention to the short-term political effects of the law. Despite this criminal-political adverse scene, we still find valuable efforts in the side of experts to bring some rationality to the legislative development. This is the case of the proposal for a new cannabis regulation model in Spain carried out by the Cannabis Policy Studies Group (hereinafter referred as ‘GEPCA’). The GEPCA is a multidisciplinary group composed by authors with multiple/different orientations, trajectories and interests, but with a common minimum objective: the conviction that the current situation regarding cannabis is unsustainable and, that a rational legislative solution must be given to the growing social pressure for the regulation of their consumption and production. This paper details the main lines through which this technical proposal is developed with the purpose of its dissemination and discussion in the Congress. The basic methodology of the proposal is inductive-expository. In that way, firstly, we will offer a brief, but solid contextualization of the situation of cannabis in Spain. This contextualization will touch on issues such as the national regulatory situation and its relationship with the international context; the criminal, judicial and penitentiary impact of the offer and consumption of cannabis, or the therapeutic use of the substance, among others. In second place, we will get down to the business properly by detailing the minutia of the three main cannabis access channels that are proposed. Namely: the regulated market, the associations of cannabis users and personal self-cultivation. In each of these options, especially in the first two, special attention will be paid to both, the production and processing of the substance and the necessary administrative control of the activity. Finally, in a third block, some notes will be given on a series of subjects that surround the different access options just mentioned above and that give fullness and coherence to the proposal outlined. Among those related issues we find some such as consumption and tenure of the substance; the issue of advertising and promotion of cannabis; consumption in areas of special risk (work or driving v. g.); the tax regime; the need to articulate evaluation instruments for the entire process; etc. The main conclusion drawn from the analysis of the proposal is the unsustainability of the current repressive system, clearly unsuccessful, and the need to develop new access routes to cannabis that guarantee both public health and the rights of people who have freely chosen to consume it.

Keywords: cannabis regulation proposal, cannabis policies studies group, criminal policy, expertise participation

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36 Food Safety in Wine: Removal of Ochratoxin a in Contaminated White Wine Using Commercial Fining Agents

Authors: Antònio Inês, Davide Silva, Filipa Carvalho, Luís Filipe-Riberiro, Fernando M. Nunes, Luís Abrunhosa, Fernanda Cosme

Abstract:

The presence of mycotoxins in foodstuff is a matter of concern for food safety. Mycotoxins are toxic secondary metabolites produced by certain molds, being ochratoxin A (OTA) one of the most relevant. Wines can also be contaminated with these toxicants. Several authors have demonstrated the presence of mycotoxins in wine, especially ochratoxin A. Its chemical structure is a dihydro-isocoumarin connected at the 7-carboxy group to a molecule of L-β-phenylalanine via an amide bond. As these toxicants can never be completely removed from the food chain, many countries have defined levels in food in order to attend health concerns. OTA contamination of wines might be a risk to consumer health, thus requiring treatments to achieve acceptable standards for human consumption. The maximum acceptable level of OTA in wines is 2.0 μg/kg according to the Commission regulation No. 1881/2006. Therefore, the aim of this work was to reduce OTA to safer levels using different fining agents, as well as their impact on white wine physicochemical characteristics. To evaluate their efficiency, 11 commercial fining agents (mineral, synthetic, animal and vegetable proteins) were used to get new approaches on OTA removal from white wine. Trials (including a control without addition of a fining agent) were performed in white wine artificially supplemented with OTA (10 µg/L). OTA analyses were performed after wine fining. Wine was centrifuged at 4000 rpm for 10 min and 1 mL of the supernatant was collected and added of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v). Also, the solid fractions obtained after fining, were centrifuged (4000 rpm, 15 min), the resulting supernatant discarded, and the pellet extracted with 1 mL of the above solution and 1 mL of H2O. OTA analysis was performed by HPLC with fluorescence detection. The most effective fining agent in removing OTA (80%) from white wine was a commercial formulation that contains gelatin, bentonite and activated carbon. Removals between 10-30% were obtained with potassium caseinate, yeast cell walls and pea protein. With bentonites, carboxymethylcellulose, polyvinylpolypyrrolidone and chitosan no considerable OTA removal was verified. Following, the effectiveness of seven commercial activated carbons was also evaluated and compared with the commercial formulation that contains gelatin, bentonite and activated carbon. The different activated carbons were applied at the concentration recommended by the manufacturer in order to evaluate their efficiency in reducing OTA levels. Trial and OTA analysis were performed as explained previously. The results showed that in white wine all activated carbons except one reduced 100% of OTA. The commercial formulation that contains gelatin, bentonite and activated carbon reduced only 73% of OTA concentration. These results may provide useful information for winemakers, namely for the selection of the most appropriate oenological product for OTA removal, reducing wine toxicity and simultaneously enhancing food safety and wine quality.

Keywords: wine, ota removal, food safety, fining

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35 Analysis of the Relationship between Micro-Regional Human Development and Brazil's Greenhouse Gases Emission

Authors: Geanderson Eduardo Ambrósio, Dênis Antônio Da Cunha, Marcel Viana Pires

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Historically, human development has been based on economic gains associated with intensive energy activities, which often are exhaustive in the emission of Greenhouse Gases (GHGs). It requires the establishment of targets for mitigation of GHGs in order to disassociate the human development from emissions and prevent further climate change. Brazil presents itself as one of the most GHGs emitters and it is of critical importance to discuss such reductions in intra-national framework with the objective of distributional equity to explore its full mitigation potential without compromising the development of less developed societies. This research displays some incipient considerations about which Brazil’s micro-regions should reduce, when the reductions should be initiated and what its magnitude should be. We started with the methodological assumption that human development and GHGs emissions arise in the future as their behavior was observed in the past. Furthermore, we assume that once a micro-region became developed, it is able to maintain gains in human development without the need of keep growing GHGs emissions rates. The human development index and the carbon dioxide equivalent emissions (CO2e) were extrapolated to the year 2050, which allowed us to calculate when the micro-regions will become developed and the mass of GHG’s emitted. The results indicate that Brazil must throw 300 GT CO2e in the atmosphere between 2011 and 2050, of which only 50 GT will be issued by micro-regions before it’s develop and 250 GT will be released after development. We also determined national mitigation targets and structured reduction schemes where only the developed micro-regions would be required to reduce. The micro-region of São Paulo, the most developed of the country, should be also the one that reduces emissions at most, emitting, in 2050, 90% less than the value observed in 2010. On the other hand, less developed micro-regions will be responsible for less impactful reductions, i.e. Vale do Ipanema will issue in 2050 only 10% below the value observed in 2010. Such methodological assumption would lead the country to issue, in 2050, 56.5% lower than that observed in 2010, so that the cumulative emissions between 2011 and 2050 would reduce by 130 GT CO2e over the initial projection. The fact of associating the magnitude of the reductions to the level of human development of the micro-regions encourages the adoption of policies that favor both variables as the governmental planner will have to deal with both the increasing demand for higher standards of living and with the increasing magnitude of reducing emissions. However, if economic agents do not act proactively in local and national level, the country is closer to the scenario in which emits more than the one in which mitigates emissions. The research highlighted the importance of considering the heterogeneity in determining individual mitigation targets and also ratified the theoretical and methodological feasibility to allocate larger share of contribution for those who historically emitted more. It is understood that the proposals and discussions presented should be considered in mitigation policy formulation in Brazil regardless of the adopted reduction target.

Keywords: greenhouse gases, human development, mitigation, intensive energy activities

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34 The Joy of Painless Maternity: The Reproductive Policy of the Bolsheviks in the 1930s

Authors: Almira Sharafeeva

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In the Soviet Union of the 1930s, motherhood was seen as a natural need of women. The masculine Bolshevik state did not see the emancipated woman as free from her maternal burden. In order to support the idea of "joyful motherhood," a medical discourse on the anesthesia of childbirth emerges. In March 1935 at the IX Congress of obstetricians and gynecologists the People's Commissar of Public Health of the RSFSR G.N. Kaminsky raised the issue of anesthesia of childbirth. It was also from that year that medical, literary and artistic editions with enviable frequency began to publish articles, studies devoted to the issue, the goal - to anesthetize all childbirths in the USSR - was proclaimed. These publications were often filled with anti-German and anti-capitalist propaganda, through which the advantages of socialism over Capitalism and Nazism were demonstrated. At congresses, in journals, and at institute meetings, doctors' discussions around obstetric anesthesia were accompanied by discussions of shortening the duration of the childbirth process, the prevention and prevention of disease, the admission of nurses to the procedure, and the proper behavior of women during the childbirth process. With the help of articles from medical periodicals of the 1930s., brochures, as well as documents from the funds of the Institute of Obstetrics and Gynecology of the Academy of Medical Sciences of the USSR (TsGANTD SPb) and the Department of Obstetrics and Gynecology of the NKZ USSR (GARF) in this paper we will show, how the advantages of the Soviet system and the socialist way of life were constructed through the problem of childbirth pain relief, and we will also show how childbirth pain relief in the USSR was related to the foreign policy situation and how projects of labor pain relief were related to the anti-abortion policy of the state. This study also attempts to answer the question of why anesthesia of childbirth in the USSR did not become widespread and how, through this medical procedure, the Soviet authorities tried to take control of a female function (childbirth) that was not available to men. Considering this subject from the perspective of gender studies and the social history of medicine, it is productive to use the term "biopolitics. Michel Foucault and Antonio Negri, wrote that biopolitics takes under its wing the control and management of hygiene, nutrition, fertility, sexuality, contraception. The central issue of biopolitics is population reproduction. It includes strategies for intervening in collective existence in the name of life and health, ways of subjectivation by which individuals are forced to work on themselves. The Soviet state, through intervention in the reproductive lives of its citizens, sought to realize its goals of population growth, which was necessary to demonstrate the benefits of living in the Soviet Union and to train a pool of builders of socialism. The woman's body was seen as the object over which the socialist experiment of reproductive policy was being conducted.

Keywords: labor anesthesia, biopolitics of stalinism, childbirth pain relief, reproductive policy

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33 Assessment of Occupational Health and Safety Conditions of Health Care Workers in Barangay Health Centers in a Selected City in Metro Manila

Authors: Deinzel R. Uezono, Vivien Fe F. Fadrilan-Camacho, Bianca Margarita L. Medina, Antonio Domingo R. Reario, Trisha M. Salcedo, Luke Wesley P. Borromeo

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The environment of health care workers is considered one of the most hazardous settings due to the nature of their work. In developing countries especially, the Philippines, this continues to be overlooked in terms of programs and services on occupational health and safety (OHS). One possible reason for this is the existing information gap on OHS which limits data comparability and impairs effective monitoring and assessment of interventions. To address this gap, there is a need to determine the current conditions of Filipino health care workers in their workplace. This descriptive cross-sectional study assessed the occupational health and safety conditions of health care workers in barangay health centers in a selected city in Metro Manila, Philippines by: (1) determining the hazards present in the workplace; (2) determining the most common self-reported medical problems; and (3) describing the elements of an OHS system based on the six building blocks of health system. Assessment was done through walkthrough survey, self-administered questionnaire, and key informant interview. Data analysis was done using Epi Info 7 and NVivo 11. Results revealed different health hazards present in the workplace particularly biological hazards (exposure to sick patients and infectious specimens), physical hazards (inadequate space and/or lighting), chemical hazards (toxic reagents and flammable chemicals), and ergonomic hazards (activities requiring repetitive motion and awkward posture). Additionally, safety hazards (improper capping of syringe and lack of fire safety provisions) were also observed. Meanwhile, the most commonly self-reported chronic diseases among health care workers (N=336) were hypertension (20.24%, n=68) and diabetes (12.50%, n=42). Top commonly self-reported symptoms were colds (66.07%, n=222), coughs (63.10%, n=212), headache (55.65%, n=187), and muscle pain (50.60%, n=170) while other diseases were influenza (16.96%, n=57) and UTI (15.48%, n=52). In terms of the elements of the OHS system, a general policy on occupational health and safety was found to be lacking and in effect, an absence of health and safety committee overseeing the implementing and monitoring of the policy. No separate budget specific for OHS programs and services was also found to be a limitation. As a result, no OHS personnel and trainings/seminar were identified. No established information system for OHS was in place. In conclusion, health and safety hazards were observed to be present across the barangay health centers visited in a selected city in Metro Manila. Medical conditions identified as most commonly self-reported were hypertension and diabetes for chronic diseases; colds, coughs, headache, and muscle pain for medical symptoms; and influenza and UTI for other diseases. As for the elements of the occupational health and safety system, there was a lack in the general components of the six building blocks of the health system.

Keywords: health hazards, occupational health and safety, occupational health and safety system, safety hazards

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32 Production of Functional Crackers Enriched with Olive (Olea europaea L.) Leaf Extract

Authors: Rosa Palmeri, Julieta I. Monteleone, Antonio C. Barbera, Carmelo Maucieri, Aldo Todaro, Virgilio Giannone, Giovanni Spagna

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In recent years, considerable interest has been shown in the functional properties of foods, and a relevant role has been played by phenolic compounds, able to scavenge free radicals. A more sustainable agriculture has to emerge to guarantee food supply over the next years. Wheat, corn, and rice are the most common cereals cultivated, but also other cereal species, such as barley can be appreciated for their peculiarities. Barley (Hordeum vulgare L.) is a C3 winter cereal that shows high resistance at drought and salt stresses. There are growing interests in barley as ingredient for the production of functional foods due to its high content of phenolic compounds and Beta-glucans. In this respect, the possibility of separating specific functional fractions from food industry by-products looks very promising. Olive leaves represent a quantitatively significant by-product of olive grove farming, and are an interesting source of phenolic compounds. In particular, oleuropein, which provide important nutritional benefits, is the main phenolic compound in olive leaves and ranges from 17% to 23% depending upon the cultivar and growing season period. Together with oleuropein and its derivatives (e.g. dimethyloleuropein, oleuropein diglucoside), olive leaves further contain tyrosol, hydroxytyrosol, and a series of secondary metabolities structurally related to them: verbascoside, ligstroside, hydroxytyrosol glucoside, tyrosol glucoside, oleuroside, oleoside-11-methyl ester, and nuzhenide. Several flavonoids, flavonoid glycosides, and phenolic acids have also described in olive leaves. The aim of this work was the production of functional food with higher content of polyphenols and the evaluation of their shelf life. Organic durum wheat and barley grains contain higher levels of phenolic compounds were used for the production of crackers. Olive leaf extract (OLE) was obtained from cv. ‘Biancolilla’ by aqueous extraction method. Two baked goods trials were performed with both organic durum wheat and barley flours, adding olive leaf extract. Control crackers, made as comparison, were produced with the same formulation replacing OLE with water. Total phenolic compound, moisture content, activity water, and textural properties at different time of storage were determined to evaluate the shelf-life of the products. Our the preliminary results showed that the enriched crackers showed higher phenolic content and antioxidant activity than control. Alternative uses of olive leaf extracts for crackers production could represent a good candidate for the addition of functional ingredients because bakery items are daily consumed, and have long shelf-life.

Keywords: barley, functional foods, olive leaf, polyphenols, shelf life

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31 Design and Biomechanical Analysis of a Transtibial Prosthesis for Cyclists of the Colombian Team Paralympic

Authors: Jhonnatan Eduardo Zamudio Palacios, Oscar Leonardo Mosquera Dussan, Daniel Guzman Perez, Daniel Alfonso Botero Rosas, Oscar Fabian Rubiano Espinosa, Jose Antonio Garcia Torres, Ivan Dario Chavarro, Ivan Ramiro Rodriguez Camacho, Jaime Orlando Rodriguez

Abstract:

The training of cilsitas with some type of disability finds in the technological development an indispensable ally, generating every day advances to contribute to the quality of life allowing to maximize the capacities of the athletes. The performance of a cyclist depends on physiological and biomechanical factors, such as aerodynamic profile, bicycle measurements, connecting rod length, pedaling systems, type of competition, among others. This study particularly focuses on the description of the dynamic model of a transtibial prosthesis for Paralympic cyclists. To make the model, two points are chosen: in the radius centers of rotation of the plate and pinion of the track bicycle. The parametric scheme of the track bike represents a model of 6 degrees of freedom due to the displacement in X - Y of each of the reference points of the angles of the curve profile β, cant of the velodrome α and the angle of rotation of the connecting rod φ. The force exerted on the crank of the bicycle varies according to the angles of the curve profile β, the velodrome cant of α and the angle of rotation of the crank φ. The behavior is analyzed through the Matlab R2015a software. The average strength that a cyclist exerts on the cranks of a bicycle is 1,607.1 N, the Paralympic cyclist must perform a force on each crank about 803.6 N. Once the maximum force associated with the movement has been determined, it is continued to the dynamic modeling of the transtibial prosthesis that represents a model of 6 degrees of freedom with displacement in X - Y in relation to the angles of rotation of the hip π, knee γ and ankle λ. Subsequently, an analysis of the kinematic behavior of the prosthesis was carried out by means of SolidWorks 2017 and Matlab R2015a, which was used to model and analyze the variation of the hip angles π, knee γ and ankle of the λ prosthesis. The reaction forces generated in the prosthesis were performed on the ankle of the prosthesis, performing the summation of forces on the X and Y axes. The same analysis was then applied to the tibia of the prosthesis and the socket. The reaction force of the parts of the prosthesis varies according to the hip angles π, knee γ and ankle of the prosthesis λ. Therefore, it can be deduced that the maximum forces experienced by the ankle of the prosthesis is 933.6 N on the X axis and 2.160.5 N on the Y axis. Finally, it is calculated that the maximum forces experienced by the tibia and the socket of the transtibial prosthesis in high performance competitions is 3.266 N on the X axis and 1.357 N on the Y axis. In conclusion, it can be said that the performance of the cyclist depends on several physiological factors, linked to biomechanics of training. The influence of biomechanical factors such as aerodynamics, bicycle measurements, connecting rod length, or non-circular pedaling systems on the cyclist performance.

Keywords: biomechanics, dynamic model, paralympic cyclist, transtibial prosthesis

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30 Co-Smoldered Digestate Ash as Additive for Anaerobic Digestion of Berry Fruit Waste: Stability and Enhanced Production Rate

Authors: Arinze Ezieke, Antonio Serrano, William Clarke, Denys Villa-Gomez

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Berry cultivation results in discharge of high organic strength putrescible solid waste which potentially contributes to environmental degradation, making it imperative to assess options for its complete management. Anaerobic digestion (AD) could be an ideal option when the target is energy generation; however, due to berry fruit characteristics high carbohydrate composition, the technology could be limited by its high alkalinity requirement which suggests dosing of additives such as buffers and trace elements supplement. Overcoming this limitation in an economically viable way could entail replacement of synthetic additives with recycled by-product waste. Consequently, ash from co-smouldering of high COD characteristic AD digestate and coco-coir could be a promising material to be used to enhance the AD of berry fruit waste, given its characteristic high pH, alkalinity and metal concentrations which is typical of synthetic additives. Therefore, the aim of the research was to evaluate the stability and process performance from the AD of BFW when ash from co-smoldered digestate and coir are supplemented as alkalinity and trace elements (TEs) source. Series of batch experiments were performed to ascertain the necessity for alkalinity addition and to see whether the alkalinity and metals in the co-smouldered digestate ash can provide the necessary buffer and TEs for AD of berry fruit waste. Triplicate assays were performed in batch systems following I/S of 2 (in VS), using serum bottles (160 mL) sealed and placed in a heated room (35±0.5 °C), after creating anaerobic conditions. Control experiment contained inoculum and substrates only, and inoculum, substrate and NaHCO3 for optimal total alkalinity concentration and TEs assays, respectively. Total alkalinity concentration refers to alkalinity of inoculum and the additives. The alkalinity and TE potential of the ash were evaluated by supplementing ash (22.574 g/kg) of equivalent total alkalinity concentration to that of the pre-determined optimal from NaHCO3, and by dosing ash (0.012 – 7.574 g/kg) of varying concentrations of specific essential TEs (Co, Fe, Ni, Se), respectively. The result showed a stable process at all examined conditions. Supplementation of 745 mg/L CaCO3 NaHCO3 resulted to an optimum TAC of 2000 mg/L CaCO3. Equivalent ash supplementation of 22.574 g/kg allowed the achievement of this pre-determined optimum total alkalinity concentration, resulting to a stable process with a 92% increase in the methane production rate (323 versus 168 mL CH4/ (gVS.d)), but a 36% reduction in the cumulative methane production (103 versus 161 mL CH4/gVS). Addition of ashes at incremental dosage as TEs source resulted to a reduction in the Cumulative methane production, with the highest dosage of 7.574 g/kg having the highest effect of -23.5%; however, the seemingly immediate bioavailability of TE at this high dosage allowed for a +15% increase in the methane production rate. With an increased methane production rate, the results demonstrated that the ash at high dosages could be an effective supplementary material for either a buffered or none buffered berry fruit waste AD system.

Keywords: anaerobic digestion, alkalinity, co-smoldered digestate ash, trace elements

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29 Microplastic Concentrations in Cultured Oyster in Two Bays of Baja California, Mexico

Authors: Eduardo Antonio Lozano Hernandez, Nancy Ramirez Alvarez, Lorena Margarita Rios Mendoza, Jose Vinicio Macias Zamora, Felix Augusto Hernandez Guzman, Jose Luis Sanchez Osorio

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Microplastics (MPs) are one of the most numerous reported wastes found in the marine ecosystem, representing one of the greatest risks for organisms that inhabit that environment due to their bioavailability. Such is the case of bivalve mollusks, since they are capable of filtering large volumes of water, which increases the risk of contamination by microplastics through the continuous exposure to these materials. This study aims to determine, quantify and characterize microplastics found in the cultured oyster Crassostrea gigas. We also analyzed if there are spatio-temporal differences in the microplastic concentration of organisms grown in two bays having quite different human population. In addition, we wanted to have an idea of the possible impact on humans via consumption of these organisms. Commercial size organisms (>6cm length; n = 15) were collected by triplicate from eight oyster farming sites in Baja California, Mexico during winter and summer. Two sites are located in Todos Santos Bay (TSB), while the other six are located in San Quintin Bay (SQB). Site selection was based on commercial concessions for oyster farming in each bay. The organisms were chemically digested with 30% KOH (w/v) and 30% H₂O₂ (v/v) to remove the organic matter and subsequently filtered using a GF/D filter. All particles considered as possible MPs were quantified according to their physical characteristics using a stereoscopic microscope. The type of synthetic polymer was determined using a FTIR-ATR microscope and using a user as well as a commercial reference library (Nicolet iN10 Thermo Scientific, Inc.) of IR spectra of plastic polymers (with a certainty ≥70% for polymers pure; ≥50% for composite polymers). Plastic microfibers were found in all the samples analyzed. However, a low incidence of MP fragments was observed in our study (approximately 9%). The synthetic polymers identified were mainly polyester and polyacrylonitrile. In addition, polyethylene, polypropylene, polystyrene, nylon, and T. elastomer. On average, the content of microplastics in organisms were higher in TSB (0.05 ± 0.01 plastic particles (pp)/g of wet weight) than found in SQB (0.02 ± 0.004 pp/g of wet weight) in the winter period. The highest concentration of MPs found in TSB coincides with the rainy season in the region, which increases the runoff from streams and wastewater discharges to the bay, as well as the larger population pressure (> 500,000 inhabitants). Otherwise, SQB is a mainly rural location, where surface runoff from streams is minimal and in addition, does not have a wastewater discharge into the bay. During the summer, no significant differences (Manne-Whitney U test; P=0.484) were observed in the concentration of MPs found in the cultured oysters of TSB and SQB, (average: 0.01 ± 0.003 pp/g and 0.01 ± 0.002 pp/g, respectively). Finally, we concluded that the consumption of oyster does not represent a risk for humans due to the low concentrations of MPs found. The concentration of MPs is influenced by the variables such as temporality, circulations dynamics of the bay and existing demographic pressure.

Keywords: FTIR-ATR, Human risk, Microplastic, Oyster

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28 Management of Urine Recovery at the Building Level

Authors: Joao Almeida, Ana Azevedo, Myriam Kanoun-Boule, Maria Ines Santos, Antonio Tadeu

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The effects of the increasing expansion of cities and climate changes have encouraged European countries and regions to adopt nature-based solutions with ability to mitigate environmental issues and improve life in cities. Among these strategies, green roofs and urban gardens have been considered ingenious solutions, since they have the desirable potential to improve air quality, prevent floods, reduce the heat island effect and restore biodiversity in cities. However, an additional consumption of fresh water and mineral nutrients is necessary to sustain larger green urban areas. This communication discusses the main technical features of a new system to manage urine recovery at the building level and its application in green roofs. The depletion of critical nutrients like phosphorus constitutes an emergency. In turn, their elimination through urine is one of the principal causes for their loss. Thus, urine recovery in buildings may offer numerous advantages, constituting a valuable fertilizer abundantly available in cities and reducing the load on wastewater treatment plants. Although several urine-diverting toilets have been developed for this purpose and some experiments using urine directly in agriculture have already been carried out in Europe, several challenges have emerged with this practice concerning collection, sanitization, storage and application of urine in buildings. To our best knowledge, current buildings are not designed to receive these systems and integrated solutions with ability to self-manage the whole process of urine recovery, including separation, maturation and storage phases, are not known. Additionally, if from a hygiene point of view human urine may be considered a relatively safe fertilizer, the risk of disease transmission needs to be carefully analysed. A reduction in microorganisms can be achieved by storing the urine in closed tanks. However, several factors may affect this process, which may result in a higher survival rate for some pathogens. In this work, urine effluent was collected under real conditions, stored in closed containers and kept in climatic chambers under variable conditions simulating cold, temperate and tropical climates. These samples were subjected to a first physicochemical and microbiological control, which was repeated over time. The results obtained so far suggest that maturation conditions were reached for all the three temperatures and that a storage period of less than three months is required to achieve a strong depletion of microorganisms. The authors are grateful for the Project WashOne (POCI-01-0247-FEDER-017461) funded by the Operational Program for Competitiveness and Internationalization (POCI) of Portugal 2020, with the support of the European Regional Development Fund (FEDER).

Keywords: sustainable green roofs and urban gardens, urban nutrient cycle, urine-based fertilizers, urine recovery in buildings

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27 Bio-Detoxification of Mycotoxins by Lactic Acid Bacteria from Different Food Matrices

Authors: António Inês, Ana Guimarães, José Maria, Vânia Laranjo, Armando Venâncio, Luís Abrunhosa

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Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP-PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 μg/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 μm pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Keywords: bio-detoxification, lactic acid bacteria, mycotoxins, food and feed

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26 Climate Change and Rural-Urban Migration in Brazilian Semiarid Region

Authors: Linda Márcia Mendes Delazeri, Dênis Antônio Da Cunha

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Over the past few years, the evidence that human activities have altered the concentration of greenhouse gases in the atmosphere have become stronger, indicating that this accumulation is the most likely cause of climate change observed so far. The risks associated with climate change, although uncertain, have the potential to increase social vulnerability, exacerbating existing socioeconomic challenges. Developing countries are potentially the most affected by climate change, since they have less potential to adapt and are those most dependent on agricultural activities, one of the sectors in which the major negative impacts are expected. In Brazil, specifically, it is expected that the localities which form the semiarid region are among the most affected, due to existing irregularity in rainfall and high temperatures, in addition to economic and social factors endemic to the region. Given the strategic limitations to handle the environmental shocks caused by climate change, an alternative adopted in response to these shocks is migration. Understanding the specific features of migration flows, such as duration, destination and composition is essential to understand the impacts of migration on origin and destination locations and to develop appropriate policies. Thus, this study aims to examine whether climatic factors have contributed to rural-urban migration in semiarid municipalities in the recent past and how these migration flows will be affected by future scenarios of climate change. The study was based on microeconomic theory of utility maximization, in which, to decide to leave the countryside and move on to the urban area, the individual seeks to maximize its utility. Analytically, we estimated an econometric model using the modeling of Fixed Effects and the results confirmed the expectation that climate drivers are crucial for the occurrence of the rural-urban migration. Also, other drivers of the migration process, as economic, social and demographic factors were also important. Additionally, predictions about the rural-urban migration motivated by variations in temperature and precipitation in the climate change scenarios RCP 4.5 and 8.5 were made for the periods 2016-2035 and 2046-2065, defined by the Intergovernmental Panel on Climate Change (IPCC). The results indicate that there will be increased rural-urban migration in the semiarid region in both scenarios and in both periods. In general, the results of this study reinforce the need for formulations of public policies to avoid migration for climatic reasons, such as policies that give support to the productive activities generating income in rural areas. By providing greater incentives for family agriculture and expanding sources of credit for the farmer, it will have a better position to face climate adversities and to settle in rural areas. Ultimately, if migration becomes necessary, there must be the adoption of policies that seek an organized and planned development of urban areas, considering migration as an adaptation strategy to adverse climate effects. Thus, policies that act to absorb migrants in urban areas and ensure that they have access to basic services offered to the urban population would contribute to the social costs reduction of climate variability.

Keywords: climate change, migration, rural productivity, semiarid region

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25 Logic of Appearance vs Explanatory Logic: A Systemic Functional Linguistics Approach to the Evolution of Communicative Strategies in the European Union Institutional Discourse

Authors: Antonio Piga

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The issue of European cultural identity has become a prominent topic of discussion among political actors in the wake of the unsuccessful referenda held in France and the Netherlands in May and June 2006. The „period of reflection‟ announced by the European Council at the conclusion of June 2006 has provided an opportunity for the implementation of several initiatives and programmes designed to „bridge the gap‟ between the EU institutions and its citizens. Specific programmes were designed with the objective of enhancing the European Commission‟s external communication of its activities. Subsequently, further plans for democracy, debate, and dialogue were devised with the objective of fostering open and extensive discourse between EU institutions and citizens. Further documentation on communication policy emphasised the necessity of developing linguistic techniques to re-engage disenchanted or uninformed citizens with the European project. It was observed that the European Union is perceived as a „faceless‟ entity, which is attributed to the absence of a distinct public identity vis-à-vis its institutions. This contribution presents an analysis of a collection of informative publications regarding the European Union, entitled “Europe on the Move”. This collection of booklets provides comprehensive information about the European Union, including its historical origins, core values, and historical development, as well as its achievements, strategic objectives, policies, and operational procedures. The theoretical framework adopted for the longitudinal linguistic analysis of EU discourse is that of Systemic Functional Linguistics (SFL). In more detail, this study considers two basic systems of relations between clauses: firstly, the degree of interdependency (or taxis) and secondly, the logico-semantic relation of expansion. The former refers to the structural markers of grammatical relations between clauses within sentences, namely paratactic, hypotactic and embedded relations. The latter pertains to various logicosemantic relationships existing between the primary and secondary members of the clause nexus. These relationships include how the secondary clause expands the primary clause, which may be achieved by (a) elaborating it, (b) extending it or (c) enhancing it. This study examines the impact of the European Commission‟s post-referendum communication methods on the portrayal of Europe, its role in facilitating the EU institutional process, and its articulation of a specific EU identity linked to distinct values. The research reveals that the language employed by the EU is evidently grounded in an explanatory logic, elucidating the rationale behind their institutionalised acts. Nevertheless, the minimal use of hypotaxis in the post-referendum booklets, coupled with the inconsistent yet increasing ratio of parataxis to hypotaxis, may suggest a potential shift towards a logic of appearance, characterised by a predominant reliance on coordination and additive, and elaborative logico-semantic relations.

Keywords: systemic functional linguistics, logic of appearance, explanatory logic, interdependency, logico-semantic relation

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24 Improvement of Greenhouse Gases Bio-Fixation by Microalgae Using a “Plasmon-Enhanced Photobioreactor”

Authors: Francisco Pereira, António Augusto Vicente, Filipe Vaz, Joel Borges, Pedro Geada

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Light is a growth-limiting factor in microalgae cultivation, where factors like spectral components, intensity, and duration, often characterized by its wavelength, are well-reported to have a substantial impact on cell growth rates and, consequently, photosynthetic performance and mitigation of CO2, one of the most significant greenhouse gases (GHGs). Photobioreactors (PBRs) are commonly used to grow microalgae under controlled conditions, but they often fail to provide an even light distribution to the cultures. For this reason, there is a pressing need for innovations aiming at enhancing the efficient utilization of light. So, one potential approach to address this issue is by implementing plasmonic films, such as the localized surface plasmon resonance (LSPR). LSPR is an optical phenomenon connected to the interaction of light with metallic nanostructures. LSPR excitation is characterized by the oscillation of unbound conduction electrons of the nanoparticles coupled with the electromagnetic field from incident light. As a result of this excitation, highly energetic electrons and a strong electromagnetic field are generated. These effects lead to an amplification of light scattering, absorption, and extinction of specific wavelengths, contingent on the nature of the employed nanoparticle. Thus, microalgae might benefit from this biotechnology as it enables the selective filtration of inhibitory wavelengths and harnesses the electromagnetic fields produced, which could lead to enhancements in both biomass and metabolite productivity. This study aimed at implementing and evaluating a “plasmon-enhanced PBR”. The goal was to utilize LSPR thin films to enhance the growth and CO2 bio-fixation rate of Chlorella vulgaris. The internal/external walls of the PBRs were coated with a TiO2 matrix containing different nanoparticles (Au, Ag, and Au-Ag) in order to evaluate the impact of this approach on microalgae’s performance. Plasmonic films with distinct compositions resulted in different Chlorella vulgaris growth, ranging from 4.85 to 6.13 g.L-1. The highest cell concentrations were obtained with the metallic Ag films, demonstrating a 14% increase compared to the control condition. Moreover, it appeared to be no differences in growth between PBRs with inner and outer wall coatings. In terms of CO2 bio-fixation, distinct rates were obtained depending on the coating applied, ranging from 0.42 to 0.53 gCO2L-1d-1. Ag coating was demonstrated to be the most effective condition for carbon fixation by C. vulgaris. The impact of LSPR films on the biochemical characteristics of biomass (e.g., proteins, lipids, pigments) was analysed as well. Interestingly, Au coating yielded the most significant enhancements in protein content and total pigments, with increments of 15 % and 173 %, respectively, when compared to the PBR without any coating (control condition). Overall, the incorporation of plasmonic films in PBRs seems to have the potential to improve the performance and efficiency of microalgae cultivation, thereby representing an interesting approach to increase both biomass production and GHGs bio-mitigation.

Keywords: CO₂ bio-fixation, plasmonic effect, photobioreactor, photosynthetic microalgae

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23 A Geographic Information System Mapping Method for Creating Improved Satellite Solar Radiation Dataset Over Qatar

Authors: Sachin Jain, Daniel Perez-Astudillo, Dunia A. Bachour, Antonio P. Sanfilippo

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The future of solar energy in Qatar is evolving steadily. Hence, high-quality spatial solar radiation data is of the uttermost requirement for any planning and commissioning of solar technology. Generally, two types of solar radiation data are available: satellite data and ground observations. Satellite solar radiation data is developed by the physical and statistical model. Ground data is collected by solar radiation measurement stations. The ground data is of high quality. However, they are limited to distributed point locations with the high cost of installation and maintenance for the ground stations. On the other hand, satellite solar radiation data is continuous and available throughout geographical locations, but they are relatively less accurate than ground data. To utilize the advantage of both data, a product has been developed here which provides spatial continuity and higher accuracy than any of the data alone. The popular satellite databases: National Solar radiation Data Base, NSRDB (PSM V3 model, spatial resolution: 4 km) is chosen here for merging with ground-measured solar radiation measurement in Qatar. The spatial distribution of ground solar radiation measurement stations is comprehensive in Qatar, with a network of 13 ground stations. The monthly average of the daily total Global Horizontal Irradiation (GHI) component from ground and satellite data is used for error analysis. The normalized root means square error (NRMSE) values of 3.31%, 6.53%, and 6.63% for October, November, and December 2019 were observed respectively when comparing in-situ and NSRDB data. The method is based on the Empirical Bayesian Kriging Regression Prediction model available in ArcGIS, ESRI. The workflow of the algorithm is based on the combination of regression and kriging methods. A regression model (OLS, ordinary least square) is fitted between the ground and NSBRD data points. A semi-variogram is fitted into the experimental semi-variogram obtained from the residuals. The kriging residuals obtained after fitting the semi-variogram model were added to NSRBD data predicted values obtained from the regression model to obtain the final predicted values. The NRMSE values obtained after merging are respectively 1.84%, 1.28%, and 1.81% for October, November, and December 2019. One more explanatory variable, that is the ground elevation, has been incorporated in the regression and kriging methods to reduce the error and to provide higher spatial resolution (30 m). The final GHI maps have been created after merging, and NRMSE values of 1.24%, 1.28%, and 1.28% have been observed for October, November, and December 2019, respectively. The proposed merging method has proven as a highly accurate method. An additional method is also proposed here to generate calibrated maps by using regression and kriging model and further to use the calibrated model to generate solar radiation maps from the explanatory variable only when not enough historical ground data is available for long-term analysis. The NRMSE values obtained after the comparison of the calibrated maps with ground data are 5.60% and 5.31% for November and December 2019 month respectively.

Keywords: global horizontal irradiation, GIS, empirical bayesian kriging regression prediction, NSRDB

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22 Polarization as a Proxy of Misinformation Spreading

Authors: Michela Del Vicario, Walter Quattrociocchi, Antonio Scala, Ana Lucía Schmidt, Fabiana Zollo

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Information, rumors, and debates may shape and impact public opinion heavily. In the latest years, several concerns have been expressed about social influence on the Internet and the outcome that online debates might have on real-world processes. Indeed, on online social networks users tend to select information that is coherent to their system of beliefs and to form groups of like-minded people –i.e., echo chambers– where they reinforce and polarize their opinions. In this way, the potential benefits coming from the exposure to different points of view may be reduced dramatically, and individuals' views may become more and more extreme. Such a context fosters misinformation spreading, which has always represented a socio-political and economic risk. The persistence of unsubstantiated rumors –e.g., the hypothetical and hazardous link between vaccines and autism– suggests that social media do have the power to misinform, manipulate, or control public opinion. As an example, current approaches such as debunking efforts or algorithmic-driven solutions based on the reputation of the source seem to prove ineffective against collective superstition. Indeed, experimental evidence shows that confirmatory information gets accepted even when containing deliberately false claims while dissenting information is mainly ignored, influences users’ emotions negatively and may even increase group polarization. Moreover, confirmation bias has been shown to play a pivotal role in information cascades, posing serious warnings about the efficacy of current debunking efforts. Nevertheless, mitigation strategies have to be adopted. To generalize the problem and to better understand social dynamics behind information spreading, in this work we rely on a tight quantitative analysis to investigate the behavior of more than 300M users w.r.t. news consumption on Facebook over a time span of six years (2010-2015). Through a massive analysis on 920 news outlets pages, we are able to characterize the anatomy of news consumption on a global and international scale. We show that users tend to focus on a limited set of pages (selective exposure) eliciting a sharp and polarized community structure among news outlets. Moreover, we find similar patterns around the Brexit –the British referendum to leave the European Union– debate, where we observe the spontaneous emergence of two well segregated and polarized groups of users around news outlets. Our findings provide interesting insights into the determinants of polarization and the evolution of core narratives on online debating. Our main aim is to understand and map the information space on online social media by identifying non-trivial proxies for the early detection of massive informational cascades. Furthermore, by combining users traces, we are finally able to draft the main concepts and beliefs of the core narrative of an echo chamber and its related perceptions.

Keywords: information spreading, misinformation, narratives, online social networks, polarization

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21 A Distributed Smart Battery Management System – sBMS, for Stationary Energy Storage Applications

Authors: António J. Gano, Carmen Rangel

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Currently, electric energy storage systems for stationary applications have known an increasing interest, namely with the integration of local renewable energy power sources into energy communities. Li-ion batteries are considered the leading electric storage devices to achieve this integration, and Battery Management Systems (BMS) are decisive for their control and optimum performance. In this work, the advancement of a smart BMS (sBMS) prototype with a modular distributed topology is described. The system, still under development, has a distributed architecture with modular characteristics to operate with different battery pack topologies and charge capacities, integrating adaptive algorithms for functional state real-time monitoring and management of multicellular Li-ion batteries, and is intended for application in the context of a local energy community fed by renewable energy sources. This sBMS system includes different developed hardware units: (1) Cell monitoring units (CMUs) for interfacing with each individual cell or module monitoring within the battery pack; (2) Battery monitoring and switching unit (BMU) for global battery pack monitoring, thermal control and functional operating state switching; (3) Main management and local control unit (MCU) for local sBMS’s management and control, also serving as a communications gateway to external systems and devices. This architecture is fully expandable to battery packs with a large number of cells, or modules, interconnected in series, as the several units have local data acquisition and processing capabilities, communicating over a standard CAN bus and will be able to operate almost autonomously. The CMU units are intended to be used with Li-ion cells but can be used with other cell chemistries, with output voltages within the 2.5 to 5 V range. The different unit’s characteristics and specifications are described, including the different implemented hardware solutions. The developed hardware supports both passive and active methods for charge equalization, considered fundamental functionalities for optimizing the performance and the useful lifetime of a Li-ion battery package. The functional characteristics of the different units of this sBMS system, including different process variables data acquisition using a flexible set of sensors, can support the development of custom algorithms for estimating the parameters defining the functional states of the battery pack (State-of-Charge, State-of-Health, etc.) as well as different charge equalizing strategies and algorithms. This sBMS system is intended to interface with other systems and devices using standard communication protocols, like those used by the Internet of Things. In the future, this sBMS architecture can evolve to a fully decentralized topology, with all the units using Wi-Fi protocols and integrating a mesh network, making unnecessary the MCU unit. The status of the work in progress is reported, leading to conclusions on the system already executed, considering the implemented hardware solution, not only as fully functional advanced and configurable battery management system but also as a platform for developing custom algorithms and optimizing strategies to achieve better performance of electric energy stationary storage devices.

Keywords: Li-ion battery, smart BMS, stationary electric storage, distributed BMS

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20 Detection and Quantification of Viable but Not Culturable Vibrio Parahaemolyticus in Frozen Bivalve Molluscs

Authors: Eleonora Di Salvo, Antonio Panebianco, Graziella Ziino

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Background: Vibrio parahaemolyticus is a human pathogen that is widely distributed in marine environments. It is frequently isolated from raw seafood, particularly shellfish. Consumption of raw or undercooked seafood contaminated with V. parahaemolyticus may lead to acute gastroenteritis. Vibrio spp. has excellent resistance to low temperatures so it can be found in frozen products for a long time. Recently, the viable but non-culturable state (VBNC) of bacteria has attracted great attention, and more than 85 species of bacteria have been demonstrated to be capable of entering this state. VBNC cells cannot grow in conventional culture medium but are viable and maintain metabolic activity, which may constitute an unrecognized source of food contamination and infection. Also V. parahaemolyticus could exist in VBNC state under nutrient starvation or low-temperature conditions. Aim: The aim of the present study was to optimize methods and investigate V. parahaemolyticus VBNC cells and their presence in frozen bivalve molluscs, regularly marketed. Materials and Methods: propidium monoazide (PMA) was integrated with real-time polymerase chain reaction (qPCR) targeting the tl gene to detect and quantify V. parahaemolyticus in the VBNC state. PMA-qPCR resulted highly specific to V. parahaemolyticus with a limit of detection (LOD) of 10-1 log CFU/mL in pure bacterial culture. A standard curve for V. parahaemolyticus cell concentrations was established with the correlation coefficient of 0.9999 at the linear range of 1.0 to 8.0 log CFU/mL. A total of 77 samples of frozen bivalve molluscs (35 mussels; 42 clams) were subsequently subjected to the qualitative (on alkaline phosphate buffer solution) and quantitative research of V. parahaemolyticus on thiosulfate-citrate-bile salts-sucrose (TCBS) agar (DIFCO) NaCl 2.5%, and incubation at 30°C for 24-48 hours. Real-time PCR was conducted on homogenate samples, in duplicate, with and without propidium monoazide (PMA) dye, and exposed for 45 min under halogen lights (650 W). Total DNA was extracted from cell suspension in homogenate samples according to bolliture protocol. The Real-time PCR was conducted with species-specific primers for V. parahaemolitycus. The RT-PCR was performed in a final volume of 20 µL, containing 10 µL of SYBR Green Mixture (Applied Biosystems), 2 µL of template DNA, 2 µL of each primer (final concentration 0.6 mM), and H2O 4 µL. The qPCR was carried out on CFX96 TouchTM (Bio-Rad, USA). Results: All samples were negative both to the quantitative and qualitative detection of V. parahaemolyticus by the classical culturing technique. The PMA-qPCR let us individuating VBNC V. parahaemolyticus in the 20,78% of the samples evaluated with a value between the Log 10-1 and Log 10-3 CFU/g. Only clams samples were positive for PMA-qPCR detection. Conclusion: The present research is the first evaluating PMA-qPCR assay for detection of VBNC V. parahaemolyticus in bivalve molluscs samples, and the used method was applicable to the rapid control of marketed bivalve molluscs. We strongly recommend to use of PMA-qPCR in order to identify VBNC forms, undetectable by the classic microbiological methods. A precise knowledge of the V.parahaemolyticus in a VBNC form is fundamental for the correct risk assessment not only in bivalve molluscs but also in other seafood.

Keywords: food safety, frozen bivalve molluscs, PMA dye, Real-time PCR, VBNC state, Vibrio parahaemolyticus

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19 From Over-Tourism to Over-Mobility: Understanting the Mobility of Incoming City Users in Barcelona

Authors: José Antonio Donaire Benito, Konstantina Zerva

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Historically, cities have been places where people from many nations and cultures have met and settled together, while population flows and density have had a significant impact on urban dynamics. Cities' high density of social, cultural, business offerings, everyday services, and other amenities not intended for tourists draw not only tourists but a wide range of city users as well. With the coordination of city rhythms and the porosity of the community, city users order and frame their urban experience. From one side, recent literature focuses on the shift in urban tourist experience from 'having' a holiday through 'doing' activities to 'becoming' a local by experiencing a part of daily life. On the other hand, there is a debate on the 'touristification of everyday life', where middle and upper class urban dwellers display attitudes and behaviors that are virtually undistinguishable from those of visitors. With the advent of globalization and technological advances, modern society has undergone a radical transformation that has altered mobility patterns within it, blurring the boundaries between tourism and everyday life, work and leisure, and "hosts" and "guests". Additionally, the presence of other 'temporary city' users, such as commuters, digital nomads, second home owners, and migrants, contributes to a more complex transformation of tourist cities. Moving away from this traditional clear distinction between 'hosts' and 'guests', which represents a more static view of tourism, and moving towards a more liquid narrative of mobility, academics on tourism development are embracing the New Mobilities Paradigm. The latter moves beyond the static structures of the modern world and focuses on the ways in which social entities are made up of people, machines, information, and images in a moving system. In light of this fluid interdependence between tourists and guests, a question arises as to whether overtourism, which is considered as the underlying cause of citizens' perception of a lower urban quality of life, is a fair representation of perceived mobility excessiveness, place consumption disruptiveness, and residents displacement. As a representative example of an overtourism narrative, Barcelona was chosen as a study area for this purpose, focusing on the incoming city users to reflect in depth the variety of people who contribute to mobility flows beyond those residents already have. Several statistical data have been analyzed to determine the number of national and international visitors to Barcelona at some point during the day in 2019. Specifically, tracking data gathered from mobile phone users within the city are combined with tourist surveys, urban mobility data, zenithal data capture, and information about the city's attractions. The paper shows that tourists are only a small part of the different incoming city users that daily enter Barcelona; excursionists, commuters, and metropolitans also contribute to a high mobility flow. Based on the diversity of incoming city users and their place consumption, it seems that the city's urban experience is more likely to be impacted by over-mobility tan over-tourism.

Keywords: city users, density, new mobilities paradigm, over-tourism.

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