Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 31

Search results for: Miceska Simona

31 Cytology Is a Promising Tool for the Diagnosis of High-Grade Serous Ovarian Carcinoma from Ascites

Authors: Miceska Simona, Škof Erik, Frković Grazio Snježana, Jeričević Anja, Smrkolj Špela, Cvjetićanin Branko, Novaković Srdjan, Grčar Kuzmanov Biljana, Kloboves-Prevodnik Veronika


Objectives: High-grade serous ovarian cancer (HGSOC) is characterized by the dissemination of the tumor cells (TC) in the peritoneal cavity forming malignant ascites at the time of diagnosis or recurrence. Still, cytology itself has been underutilized as a modality for the diagnosis of HGSOC from ascites, and histological examination from the tumor tissue is yet the only validated method used. The objective of this study was to evaluate the reliability of cytology in the diagnosis of HGSOC in relation to the histopathological examination. Methods: The study included 42 patients with histologically confirmed HGSOC, accompanied by malignant ascites. To confirm the malignancy of the TC in the ascites and to define their immunophenotype, immunohistochemical reaction (IHC) of the following antigens: Calretinin, MOC, WT1, PAX8, p53, p16 & Ki-67 was evaluated on ascites cytospins and tissue blocks. For complete cytological determination of HGSOC, BRCA 1/2 gene mutation was determined from ascites, tissue block, and blood. BRCA1/2 mutation from blood was performed to define the type of mutation, somatic vs germline. Results: Among 42 patients, the immunophenotype of HGSOC from ascites was confirmed in 36 cases (86%). For more profound analysis, the patients were divided in 3 groups regarding the number of TC present in the ascites: patients with less than 10% TC, 10% TC, and more than 10% TC. From all included patients, in the group with less than 10% TC, there were 10 cases, and only 5 of them(50%) showed HGSOC phenotype; 12 cases had equally 10% of TC, and 11 cases (92%) showed HGSOC phenotype; 20 cases had more than 10% TC and all of them (100%) confirmed the HGSOC immunophenotype from ascites. Only 33 patients were eligible for further BRCA1/2 analysis. Eleven BRCA1/2 mutations were detected from thetissue block: 6 germline and 5 somatic. In 2 cases with less than 10% TC, BRCA1/2 mutation was not detected; 4 cases had 10% TC, and 2 of them (50%) confirmed the mutation; 4 cases had more than 10% TC, and all showed 100% reliability with the tumor tissue. Conclusions: Cytology is a highly reliable method for determining the immunophenotype of HGSOC and BRCA1/2 mutation if more than 10% of tumor cells are present in the ascites. This may present an additional non-invasive clinical approach for fast and effective diagnose in the future, especially in inoperable conditions or relapses.

Keywords: cytology, ascites, high-grade serous ovarian cancer, immunophenotype, BRCA1/2

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30 The Effects of a Thin Liquid Layer on the Hydrodynamic Machine Rotor

Authors: Jaroslav Krutil, František Pochylý, Simona Fialová, Vladimír Habán


A mathematical model of the additional effects of the liquid in the hydrodynamic gap is presented in the paper. An in-compressible viscous fluid is considered. Based on computational modeling are determined the matrices of mass, stiffness and damping. The mathematical model is experimentally verified.

Keywords: computational modeling, mathematical model, hydrodynamic gap, matrices of mass, stiffness and damping

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29 Nonlinear Mathematical Model of the Rotor Motion in a Thin Hydrodynamic Gap

Authors: Jaroslav Krutil, Simona Fialová, , František Pochylý


A nonlinear mathematical model of mutual fluid-structure interaction is presented in the work. The model is applicable to the general shape of sealing gaps. An in compressible fluid and turbulent flow is assumed. The shaft carries a rotational and procession motion, the gap is axially flowed through. The achieved results of the additional mass, damping and stiffness matrices may be used in the solution of the rotor dynamics. The usage of this mathematical model is expected particularly in hydraulic machines. The method of control volumes in the ANSYS Fluent was used for the simulation. The obtained results of the pressure and velocity fields are used in the mathematical model of additional effects.

Keywords: nonlinear mathematical model, CFD modeling, hydrodynamic sealing gap, matrices of mass, stiffness, damping

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28 The Impact of Corporate Governance on Risk Taking in European Insurance Industry

Authors: Francesco Venuti, Simona Alfiero


The aim of this paper is to develop an empirical research on the nature and consequences of corporate governance on Eurozone Insurance Industry risk taking attitude. More particularly, we analyzed the effect of public ownership on risk taking with respect to privately held Insurance Companies. We also analyzed the effects on risk taking attitude of different degrees of ownership concentration, directors compensation, and the dimension/diversity of the Board of Directors. Our results provide quite strong evidence that, coherently with the Agency Theory, publicly traded insurance companies with more concentrated ownership are less risky than the corresponding privately held.

Keywords: agency theory, corporate governance, insurance companies, risk taking

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27 Generalized Rough Sets Applied to Graphs Related to Urban Problems

Authors: Mihai Rebenciuc, Simona Mihaela Bibic


Branch of modern mathematics, graphs represent instruments for optimization and solving practical applications in various fields such as economic networks, engineering, network optimization, the geometry of social action, generally, complex systems including contemporary urban problems (path or transport efficiencies, biourbanism, & c.). In this paper is studied the interconnection of some urban network, which can lead to a simulation problem of a digraph through another digraph. The simulation is made univoc or more general multivoc. The concepts of fragment and atom are very useful in the study of connectivity in the digraph that is simulation - including an alternative evaluation of k- connectivity. Rough set approach in (bi)digraph which is proposed in premier in this paper contribute to improved significantly the evaluation of k-connectivity. This rough set approach is based on generalized rough sets - basic facts are presented in this paper.

Keywords: (bi)digraphs, rough set theory, systems of interacting agents, complex systems

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26 The MSC-MCF-7 Duet Playing Tumor Vasculogenesis and Angiogenesis Onto the Chick Embryo Chorioallantoic Membrane

Authors: Serban Comsa, Amalia-Raluca Ceausu, Roxana Popescu, Simona Sarb, Anca-Maria Cimpean, Marius Raica


Background/Aim: The human mesenchymal stem cells (hMSC) represent a versatile cell population, able to modulate the tumor microenvironment Our aim was to recreate an open scene for the in vivo interaction between hMSC and the MCF-7 breast cancer cells (MCF-7), in order to enlighten the intimate involvement of hMSC in tumor vasculogenesis and angiogenesis. Materials and Methods: hMSC and MCF-7 were seeded onto the chick embryo chorioallantoic membrane (CAM) and incubated for seven days. Consecutively, the morphology and the immunohistochemical profile of CAM were assessed. Results: Following this complex interaction, MCF-7 acquired a more aggressive phenotype, hMSC switched to a vascular precursor phenotype whileCAM underwent a major reset to an earlier stage, with hotspots of angiogenesis, vasculogenesis and hematopoiesis. Conclusion: The hallmark of this study was the establishment of a veritable in vivo experimental model of MSC involvement in tumor vasculogenesis and angiogenesis, allowing further analysis in the field.

Keywords: angiogenesis, chorioallantoic membrane, MCF-7, mesenchymal stem cells, vasculogenesis

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25 Games behind Bars: A Longitudinal Study of Inmates Pro-Social Preferences

Authors: Mario A. Maggioni, Domenico Rossignoli, Simona Beretta, Sara Balestri


The paper presents the results of a Longitudinal Randomized Control Trial implemented in 2016 two State Prisons in California (USA). The subjects were randomly assigned to a 10-months program (GRIP, Guiding Rage Into Power) aiming at undoing the destructive behavioral patterns that lead to criminal actions by raising the individual’s 'mindfulness'. This study tests whether the participation to this program (treatment), based on strong relationships and mutual help, affects pro-social behavior of participants, in particular with reference to trust and inequality aversion. The research protocol entails the administration of two questionnaires including a set of behavioral situations ('games') - widely used in the relevant literature in the field - to 80 inmates, 42 treated (enrolled in the program) and 38 controls. The first questionnaire has been administered before treatment and randomization took place; the second questionnaire at the end of the program. The results of a Difference-in-Differences estimation procedure, show that trust significantly increases GRIP participants to compared to the control group. The result is robust to alternative estimation techniques and to the inclusion of a set of covariates to further control for idiosyncratic characteristics of the prisoners.

Keywords: behavioral economics, difference in differences, longitudinal study, pro-social preferences

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24 In vitro Effects of Viscum album on the Functionality of Rabbit Spermatozoa

Authors: Marek Halenár, Eva Tvrdá, Simona Baldovská, Ľubomír Ondruška, Peter Massányi, Adriana Kolesárová


This study aimed to assess the in vitro effects of different concentrations of the Viscum album extract on the motility, viability, and reactive oxygen species (ROS) production by rabbit spermatozoa during different time periods (0, 2, and 8h). Spermatozoa motility was assessed by using the CASA (Computer aided sperm analysis) system. Cell viability was evaluated by using the metabolic activity MTT assay, and the luminol-based luminometry was applied to quantify the ROS formation. The CASA analysis revealed that low Viscum concentrations were able to prevent a rapid decline of spermatozoa motility, especially in the case of concentrations ranging between 1 and 5 µg/mL (P<0.05 with respect to time 8h). At the same time, concentrations ranging between 1 and 100 µg/mL of the extract led to a significant preservation of the cell viability (P<0.05 in case of 5, 50 and 100 µg/mL; P<0.01 with respect to 1 and 10 µg/mL, time 8h). 1 and 5 µg/mL of the extract exhibited antioxidant characteristics, translated into a significant reduction of the ROS production, particularly notable at time 8h (P<0.01). The results indicate that the Viscum extract is capable of delaying the damage inflicted to the spermatozoon by the in vitro environment.

Keywords: CASA, mistletoe, mitochondrial activity, motility, reactive oxygen species, rabbits, spermatozoa, Viscum album

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23 RF Plasma Discharge Equipment for Conservation Treatments of Paper Supports

Authors: Emil Ghiocel Ioanid, Viorica Frunză, Dorina Rusu, Ana Maria Vlad, Catalin Tanase, Simona Dunca


The application of cold radio-frequency (RF) plasma in the conservation of cultural heritage became important in the last decades due to the positive results obtained in decontamination treatments. This paper presents an equipment especially designed for RF cold plasma application on paper documents, developed within a research project. The equipment allows the application of decontamination and cleaning treatments on any type of paper support, as well as the coating with a protective polymer. The equipment consists in a Pyrex vessel, inside which are placed two plane-parallel electrodes, capacitively coupled to a radio-frequency generator. The operating parameters of the equipment are: 1.2 MHz frequency, 50V/cm electric field intensity, current intensity in the discharge 100 mA, 40 W power in the discharge, the pressure varying from 5∙10-1 mbar to 5.5∙10-1 mbar, depending on the fragility of the material, operating in gaseous nitrogen. In order to optimize the equipment treatments in nitrogen plasma have been performed on samples infested with microorganisms, then the decontamination and the changes in surface properties (color, pH) were assessed. The analyses results presented in the table revealed only minor modifications of surface pH the colorimetric analysis showing a slight change to yellow. The equipment offers the possibility of performing decontamination, cleaning and protective coating of paper-based documents in successive stages, thus avoiding the recontamination with harmful biological agents.

Keywords: nitrogen plasma, cultural heritage, paper support, radio-frequency

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22 Feasibilities for Recovering of Precious Metals from Printed Circuit Board Waste

Authors: Simona Ziukaite, Remigijus Ivanauskas, Gintaras Denafas


Market development of electrical and electronic equipment and a short life cycle is driven by the increasing waste streams. Gold Au, copper Cu, silver Ag and palladium Pd can be found on printed circuit board. These metals make up the largest value of printed circuit board. Therefore, the printed circuit boards scrap is valuable as potential raw material for precious metals recovery. A comparison of Cu, Au, Ag, Pd recovery from waste printed circuit techniques was selected metals leaching of chemical reagents. The study was conducted using the selected multistage technique for Au, Cu, Ag, Pd recovery of printed circuit board. In the first and second metals leaching stages, as the elution reagent, 2M H2SO4 and H2O2 (35%) was used. In the third stage, leaching of precious metals used solution of 20 g/l of thiourea and 6 g/l of Fe2 (SO4)3. Verify the efficiency of the method was carried out the metals leaching test with aqua regia. Based on the experimental study, the leaching efficiency, using the preferred methodology, 60 % of Au and 85,5 % of Cu dissolution was achieved. Metals leaching efficiency after waste mechanical crushing and thermal treatment have been increased by 1,7 times (40 %) for copper, 1,6 times (37 %) for gold and 1,8 times (44 %) for silver. It was noticed that, the Au amount in old (> 20 years) waste is 17 times more, Cu amount - 4 times more, and Ag - 2 times more than in the new (< 1 years) waste. Palladium in the new printed circuit board waste has not been found, however, it was established that from 1 t of old printed circuit board waste can be recovered 1,064 g of Pd (leaching with aqua regia). It was found that from 1 t of old printed circuit board waste can be recovered 1,064 g of Ag. Precious metals recovery in Lithuania was estimated in this study. Given the amounts of generated printed circuit board waste, the limits for recovery of precious metals were identified.

Keywords: leaching efficiency, limits for recovery, precious metals recovery, printed circuit board waste

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21 African Swine Fewer Situation and Diagnostic Methods in Lithuania

Authors: Simona Pileviciene


On 24th January 2014, Lithuania notified two primary cases of African swine fever (ASF) in wild boars. The animals were tested positive for ASF virus (ASFV) genome by real-time PCR at the National Reference Laboratory for ASF in Lithuania (NRL), results were confirmed by the European Union Reference Laboratory for African swine fever (CISA-INIA). Intensive wild and domestic animal monitoring program was started. During the period of 2014-2017 ASF was confirmed in two large commercial pig holding with the highest biosecurity. Pigs were killed and destroyed. Since 2014 ASF outbreak territory from east and south has expanded to the middle of Lithuania. Diagnosis by PCR is one of the highly recommended diagnostic methods by World Organization for Animal Health (OIE) for diagnosis of ASF. The aim of the present study was to compare singleplex real-time PCR assays to a duplex assay allowing the identification of ASF and internal control in a single PCR tube and to compare primers, that target the p72 gene (ASF 250 bp and ASF 75 bp) effectivity. Multiplex real-time PCR assays prove to be less time consuming and cost-efficient and therefore have a high potential to be applied in the routine analysis. It is important to have effective and fast method that allows virus detection at the beginning of disease for wild boar population and in outbreaks for domestic pigs. For experiments, we used reference samples (INIA, Spain), and positive samples from infected animals in Lithuania. Results show 100% sensitivity and specificity.

Keywords: African swine fewer, real-time PCR, wild boar, domestic pig

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20 Strain Sensing Seams for Monitoring Body Movement

Authors: Sheilla Atieno Odhiambo, Simona Vasile, Alexandra De Raeve, Ann Schwarz


Strain sensing seams have been developed by integrating conductive sewing threads in different types of seams design on a fabric typical for sports clothing using sewing technology. The aim is to have a simple integrated textile strain sensor that can be applied to sports clothing to monitor the movements of the upper body parts of the user during sports. Different types of commercially available sewing threads were used as the bobbin thread in the production of different architectural seam sensors. These conductive sewing threads have been integrated into seams in particular designs using specific seam types. Some of the threads are delicate and needed to be laid into the seam with as little friction as possible and less tension; thus, they could only be sewn in as the bobbin thread and not the needle thread. Stitch type 304; 406; 506; 601;602; 605. were produced. The seams were made on a fabric of 80% polyamide 6.6 and 20% elastane. The seams were cycled(stretch-release-stretch) for five cycles and up to 44 cycles following EN ISO 14704-1: 2005 (modified), using a tensile instrument and the changes in the resistance of the seams with time were recorded using Agilent meter U1273A. Both experiments were conducted simultaneously on the same seam sample. Sensing functionality, among which is sensor gauge and reliability, were evaluated on the promising sensor seams. The results show that the sensor seams made from HC Madeira 40 conductive yarns performed better inseam stitch 304 and 602 compared to the other combination of stitch type and conductive sewing threads. These sensing seams 304, 406 and 602 will further be interconnected to our developed processing and communicating unit and further integrated into a sports clothing prototype that can track body posture. This research is done within the framework of the project SmartSeam.

Keywords: conductive sewing thread, sensing seams, smart seam, sewing technology

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19 Raman, Atomic Force Microscopy and Mass Spectrometry for Isotopic Ratios Methods Used to Investigate Human Dentine and Enamel

Authors: Nicoleta Simona Vedeanu, Rares Stiufiuc, Dana Alina Magdas


A detailed knowledge of the teeth structure is mandatory to understand and explain the defects and the dental pathology, but especially to take a correct decision regarding dental prophylaxis and treatment. The present work is an alternative study to the traditional investigation methods used in dentistry, a study based on the use of modern, sensitive physical methods to investigate human enamel and dentin. For the present study, several teeth collected from patients of different ages were used for structural and dietary investigation. The samples were investigated by Raman spectroscopy for the molecular structure analysis of dentin and enamel, atomic force microscopy (AFM) to view the dental topography at the micrometric size and mass spectrometry for isotopic ratios as a fingerprint of patients’ personal diet. The obtained Raman spectra and their interpretation are in good correlation with the literature and may give medical information by comparing affected dental structures with healthy ones. AFM technique gave us the possibility to study in details the dentin and enamel surface to collect information about dental hardness or dental structural changes. δ¹³C values obtained for the studied samples can be classified in C4 category specific to young people and children diet (sweets, cereals, juices, pastry). The methods used in this attempt furnished important information about dentin and enamel structure and dietary habits and each of the three proposed methods can be extended at a larger level in the study of the teeth structure.

Keywords: AFM, dentine, enamel, Raman spectroscopy

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18 Drape Simulation by Commercial Software and Subjective Assessment of Virtual Drape

Authors: Evrim Buyukaslan, Simona Jevsnik, Fatma Kalaoglu


Simulation of fabrics is more difficult than any other simulation due to complex mechanics of fabrics. Most of the virtual garment simulation software use mass-spring model and incorporate fabric mechanics into simulation models. The accuracy and fidelity of these virtual garment simulation software is a question mark. Drape is a subjective phenomenon and evaluation of drape has been studied since 1950’s. On the other hand, fabric and garment simulation is relatively new. Understanding drape perception of subjects when looking at fabric simulations is critical as virtual try-on becomes more of an issue by enhanced online apparel sales. Projected future of online apparel retailing is that users may view their avatars and try-on the garment on their avatars in the virtual environment. It is a well-known fact that users will not be eager to accept this innovative technology unless it is realistic enough. Therefore, it is essential to understand what users see when they are displaying fabrics in a virtual environment. Are they able to distinguish the differences between various fabrics in virtual environment? The purpose of this study is to investigate human perception when looking at a virtual fabric and determine the most visually noticeable drape parameter. To this end, five different fabrics are mechanically tested, and their drape simulations are generated by commercial garment simulation software (Optitex®). The simulation images are processed by an image analysis software to calculate drape parameters namely; drape coefficient, node severity, and peak angles. A questionnaire is developed to evaluate drape properties subjectively in a virtual environment. Drape simulation images are shown to 27 subjects and asked to rank the samples according to their questioned drape property. The answers are compared to the calculated drape parameters. The results show that subjects are quite sensitive to drape coefficient changes while they are not very sensitive to changes in node dimensions and node distributions.

Keywords: drape simulation, drape evaluation, fabric mechanics, virtual fabric

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17 The Role of the STAT3 Signaling for Melatonergic Synthetic Pathway in the Rat Pineal Gland

Authors: Simona Moravcova, Jiri Novotny, Zdenka Bendova


The pineal gland of the vertebrate brain is a circumventricular organ which serves as a major neuroendocrine gland with the primary function of rhythmic secretion of neurohormone melatonin under the control of the hypothalamic suprachiasmatic nucleus (SCN). Soon after the onset of the darkness, the activity of the key rate-limiting enzyme for melatonin synthesis, arylalkylamine N-acetyltransferase (AANAT), raises due to the increased release of norepinephrine from sympathetic neurons terminating on the parenchymal cells where it binds to β-adrenergic receptors. Melatonin codes the length of the night, and it is well recognized for its anti-inflammatory effects. However, to our knowledge, less is known about the effect of the immune system on the melatonin biosynthesis and the precise role of the STAT3 in the signaling pathway leading to the expression of AANAT. Lipopolysaccharide (LPS) is the essential component in the outer surface membrane of gram-negative bacteria and acts as a strong stimulator of natural and innate immunity. STAT3 acts as an important factor in immune response. Here we investigated the effect of LPS on the components of the melatonergic synthetic pathway in the pineal gland. The experiments were performed both in vivo and in vitro. The changes in AANAT activity were determined by radioenzymatic assay. PCR analyses were carried out to detect aa-nat, icer, spi-3 and stat3 gene expression. From our results, it is apparent that the high basal level of phosphorylated forms of STAT3 can be elevated after systemic as well as in vitro administration of LPS. Our experiments have shown that LPS reduces melatonin synthesis, nevertheless, the activity of AANAT was increased. Moreover, the basal level of phosphorylated STAT3 counteracts β-adrenergic receptor-mediated aa-nat gene expression and sustains its own and spi-3 gene expression. In conclusion, LPS can affect immunomodulators such as melatonin in the pineal gland.

Keywords: AANAT, lipopolysaccharide, pineal gland, rat, STAT3

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16 White Wine Discrimination Based on Deconvoluted Surface Enhanced Raman Spectroscopy Signals

Authors: Dana Alina Magdas, Nicoleta Simona Vedeanu, Ioana Feher, Rares Stiufiuc


Food and beverages authentication using rapid and non-expensive analytical tools represents nowadays an important challenge. In this regard, the potential of vibrational techniques in food authentication has gained an increased attention during the last years. For wines discrimination, Raman spectroscopy appears more feasible to be used as compared with IR (infrared) spectroscopy, because of the relatively weak water bending mode in the vibrational spectroscopy fingerprint range. Despite this, the use of Raman technique in wine discrimination is in an early stage. Taking this into consideration, the wine discrimination potential of surface-enhanced Raman scattering (SERS) technique is reported in the present work. The novelty of this study, compared with the previously reported studies, concerning the application of vibrational techniques in wine discrimination consists in the fact that the present work presents the wines differentiation based on the individual signals obtained from deconvoluted spectra. In order to achieve wines classification with respect to variety, geographical origin and vintage, the peaks intensities obtained after spectra deconvolution were compared using supervised chemometric methods like Linear Discriminant Analysis (LDA). For this purpose, a set of 20 white Romanian wines from different viticultural Romanian regions four varieties, was considered. Chemometric methods applied directly to row SERS experimental spectra proved their efficiency, but discrimination markers identification found to be very difficult due to the overlapped signals as well as for the band shifts. By using this approach, a better general view related to the differences that appear among the wines in terms of compositional differentiation could be reached.

Keywords: chemometry, SERS, variety, wines discrimination

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15 Intentional Cultivation of Non-toxic Filamentous Cyanobacteria Tolypothrix as an Approach to Treat Eutrophic Waters

Authors: Simona Lucakova, Irena Branyikova


Eutrophication, a condition when water becomes over-enriched with nutrients (P, N), can lead to undesirable excessive growth of phytoplankton, so-called algal bloom. This process results in the accumulation of toxin-producing cyanobacteria and oxygen depletion, both possibly leading to the collapse of the whole ecosystem. In real conditions, the limiting nutrient, which determines the possible growth of harmful algal bloom, is usually phosphorus. Algicides or flocculants have been applied in the eutrophicated waterbody in order to reduce the phytoplankton growth, which leads to the introduction of toxic chemicals into the water. In our laboratory, the idea of the prevention of harmful phytoplankton growth by the intentional cultivation of non-toxic cyanobacteria Tolypothrix tenuis in semi-open floating photobioreactors directly on the surface of phosphorus-rich waterbody is examined. During the process of cultivation, redundant phosphorus is incorporated into cyanobacterial biomass, which can be subsequently used for the production of biofuels, cosmetics, pharmaceuticals, or biostimulants for agricultural use. To determine the ability of phosphorus incorporation, batch-cultivation of Tolypothrix biomass in media simulating eutrophic water (10% BG medium) and in effluent from municipal wastewater treatment plant, both with the initial phosphorus concentration in the range 0.5-1.0 mgP/L was performed in laboratory-scale models of floating photobioreactors. After few hours of cultivation, the phosphorus content was decreased below the target limit of 0.035 mgP/L, which was given as a borderline for the algal bloom formation. Under laboratory conditions, the effect of several parameters on the rate of phosphorus decrease was tested (illumination, temperature, stirring speed/aeration gas flow, biomass to medium ratio). Based on the obtained results, a bench-scale floating photobioreactor was designed and will be tested for Tolypothrix growth in real conditions. It was proved that intentional cultivation of cyanobacteria Tolypothrix could be a suitable approach for extracting redundant phosphorus from eutrophic waters as prevention of algal bloom formation.

Keywords: cyanobacteria, eutrophication, floating photobioreactor, Tolypothrix

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14 Extracellular Enzymes from Halophilic Bacteria with Potential in Agricultural Secondary Flow Recovery Products

Authors: Madalin Enache, Simona Neagu, Roxana Cojoc, Ioana Gomoiu, Delia Ionela Dobre, Ancuta Roxana Trifoi


Various types of halophilic and halotolerant microorganisms able to be cultivated in laboratory on culture media with a wide range of sodium chloride content are isolated from several salted environments. The extracellular enzymes of these microorganisms showed the enzymatic activity in these spectrums of salinity thus being attractive for several biotechnological processes developed at high ionic strength. In present work, a number of amylase, protease, esterase, lipase, cellulase, pectinase, xilanases and innulinase were identified for more than 50th bacterial strains isolated from water samples and sapropelic mud from four saline and hypersaline lakes located in Romanian plain. On the other hand, the cellulase and pectinase activity were also detected in some halotolerant microorganisms isolated from secondary agricultural flow of grapes processing. The preliminary data revealed that from totally tested strains seven harbor proteases activity, eight amylase activity, four for esterase and another four for lipase, three for pectinase and for one strain were identified either cellulase or pectinase activity. There were no identified enzymes able to hydrolase innulin added to culture media. Several strains isolated from sapropelic mud showed multiple extracellular enzymatic activities, namely three strains harbor three activities and another seven harbor two activities. The data revealed that amylase and protease activities were frequently detected if compare with other tested enzymes. In the case of pectinase were investigated, their ability to be used for increasing resveratrol recovery from material resulted after grapes processing. In this way, the resulted material from grapes processing was treated with microbial supernatant for several times (two, four and 24 hours) and the content of resveratrol was detected by High Performance Liquid Chromatography method (HPLC). The preliminary data revealed some positive results of this treatment.

Keywords: halophilic microorganisms, enzymes, pectinase, salinity

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13 Therapeutic Efficacy of Clompanus Pubescens Leaves Fractions via Downregulation of Neuronal Cholinesterases/NA⁺-K⁺ ATPase/IL-1 β and Improving the Neurocognitive and Antioxidants Status of Streptozotocin-Induced Diabetic Rats

Authors: Amos Sunday Onikanni, Bashir Lawal, Babatunji Emmanuel Oyinloye, Gomaa Mostafa-Hedeab, Mohammed Alorabi, Simona Cavalu, Augustine O. Olusola, Chih-Hao Wang, Gaber El-Saber Batiha


The increasing global burden of diabetes mellitus has called for the search for a therapeutic alternative that offers better activities and safety than conventional chemotherapy. Herein, we evaluated the neuroprotective and antioxidant properties of different fractions (ethyl acetate, N-butanol and residual aqueous) of Clompanus pubescens leaves in streptozotocin (STZ)-induced diabetic rats. Our results revealed a significant elevation in the levels of blood glucose, pro-inflammatory cytokines, lipid peroxidation, neuronal activities of acetylcholinesterase, butyrylcholinesterase, nitric oxide, epinephrine, norepinephrine, and Na+/K+-ATPase in diabetic non treated rats. In addition, decreased levels of enzymatic and non-enzymatic antioxidants were observed. Treatment with different fractions of C. pubescens leaves resulted in a significant reversal of the biochemical alteration and improved the neurocognitive deficit in STZ-induced diabetic rats. However, the ethyl-acetate fraction demonstrated higher activities than the other fractions and was characterized for its phytoconstituents, revealing the presence of Gallic acid (713.00 ppm), catechin (0.91 ppm), ferulic acid (0.98 ppm), rutin (59.82 ppm), quercetin (3.22 ppm) and kaempferol (4.07 ppm). Our molecular docking analysis revealed that these compounds exhibited different binding affinities and potentials for targeting BChE/AChE/ IL-1 β/Na+-K+-ATPase. However, only Kampferol and ferulic exhibited good drug-like, ADMET, and permeability properties suitable for use as a neuronal drug target agent. Hence, the ethyl-acetate fraction of C. pubescent leaves could be considered a source of promising bioactive metabolite for the treatment and management of cognitive impairments related to type II diabetes mellitus.

Keywords: diabetes mellitus, neuroprotective, antioxidant, pro-inflammatory cytokines

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12 Effect of Texturised Soy Protein and Yeast on the Instrumental and Sensory Quality of Hybrid Beef Meatballs

Authors: Simona Grasso, Gabrielle Smith, Sophie Bowers, Oluseyi Moses Ajayi, Mark Swainson


Hybrid meat analogues are meat products whereby a proportion of meat has been partially replaced by more sustainable protein sources. These products could bridge the gap between meat and meat-free products, providing convenience, and allowing consumers to continue using meat products as they conventionally would, while lowering their overall meat intake. The study aimed to investigate the effect of introducing texturized soy protein (TSP) at different levels (15% and 30%) with and without nutritional yeast as flavour enhancer on the sensory and instrumental quality of beef meatballs, compared to a soy and yeast-free control. Proximate analysis, yield, colour, instrumental texture, and sensory quality were investigated. The addition of soy and yeast did not have significant effects on the overall protein content, but the total fat and moisture content went down with increasing soy substitution. Samples with 30% TSP had significantly higher yield than the other recipes. In terms of colour, a* redness values tended to go down and b* yellowness values tended to go up with increasing soy addition. The addition of increasing levels of soy and yeast modified the structure of meatballs resulting in a progressive decrease in hardness and chewiness compared to control. Sixty participants assessed the samples using Check-all-that-apply (CATA) questions and hedonic scales. The texture of all TSP-containing samples received significantly higher acceptability scores than control, while 15% TSP with yeast received significantly higher flavour and overall acceptability scores than control. Control samples were significantly more often associated than the other recipes to the term 'hard' and the least associated to 'soft' and 'crumbly and easy to cut'. All recipes were similarly associated to the terms 'weak meaty', 'strong meaty', 'characteristic' and 'unusual'. Correspondence analysis separated the meatballs in three distinct groups: 1) control; 2) 30%TSP with yeast; and 3) 15%TSP, 15%TSP with yeast and 30%TSP located together on the sensory map, showing similarity. Adding 15-30% TSP with or without yeast inclusion could be beneficial for the development of future meat hybrids with acceptable sensory quality. These results can provide encouragement for the use of the hybrid concept by the meat industry to promote the partial substitution of meat in flexitarians’ diets.

Keywords: CATA, hybrid meat products, texturised soy protein, yeast

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11 Phage Capsid for Efficient Delivery of Cytotoxic Drugs

Authors: Simona Dostalova, Dita Munzova, Ana Maria Jimenez Jimenez, Marketa Vaculovicova, Vojtech Adam, Rene Kizek


The boom of nanomedicine in recent years has led to the development of numerous new nanomaterials that can be used as nanocarriers in the drug delivery. These nanocarriers can either be synthetic or natural-based. The disadvantage of many synthetic nanocarriers is their toxicity in patient’s body. Protein cages that can naturally be found in human body do not exhibit such disadvantage. However, the release of cargo from some protein cages in target cells can be problematic. As a special type of protein cages can serve the capsid of many viruses, including phage. Phages infect bacterial cells; therefore they are not harmful to human cells. The targeting of phage particles to cancer cells can be solved by producing of empty phage capsids during which the targeting moieties (e.g. peptides) can be cloned into genes of phage capsid to decorate its surface. Moreover, the produced capsids do not contain viral nucleic acid and are therefore not infectious to beneficial bacteria in the patient’s body. The protein cage composed of viral capsid is larger than other frequently used apoferritin cage but its size is still small enough to benefit from passive targeting by Enhanced Permeability and Retention effect. In this work, bacteriophage λ was used, both whole and its empty capsid for delivery of different cytotoxic drugs (cisplatin, carboplatin, oxaliplatin, etoposide and doxorubicin). Large quantities of phage λ were obtained from phage λ-producing strain of E. coli cultivated in medium with 0.2 % maltose. After killing of E. coli with chloroform and its removal by centrifugation, the phage was concentrated by ultracentrifugation at 130 000 g and 4 °C for 3 h. The encapsulation of the drugs was performed by infusion method and four different concentrations of the drugs were encapsulated (200; 100; 50; 25 µg/ml). Free molecules of drugs were removed by dialysis. The encapsulation was verified using spectrophotometric and electrochemical methods. The amount of encapsulated drug linearly increased with the amount of applied drug (determination coefficient R2=0.8013). 76% of applied drug was encapsulated in phage λ particles (concentration of 10 µg/ml), even with the highest applied concentration of drugs, 200 µg/ml. Only 1% of encapsulated drug was detected in phage DNA. Similar results were obtained with encapsulation in phage empty capsid. Therefore, it can be concluded that the encapsulation of drugs into phage particles is efficient and mostly occurs by interaction of drugs with protein capsid.

Keywords: cytostatics, drug delivery, nanocarriers, phage capsid

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10 Managing of Cobalt and Chromium Ions by Patients with Metal-on-Metal Hip Prosthesis

Authors: Alina Beraudi, Simona Catalani, Dalila De Pasquale, Eva Bianconi, Umberto Santoro, Susanna Stea, Pietro Apostoli


Recently the European Community, in line with the international scientific community such as with the Consensus Statement, has determined to stop the use of metal-on-metal big head stemmed hip prosthesis. Among the factors accounted as responsible for the high failure rates of these hip implants are the release and accumulation of metal ions. Many studies have correlated the presence of these ions, besides other factors, with the induction of oxidative stress response. In our study on 12 subjects, we observed the patient specific capability to eliminate metal ions after revision surgery. While for cobalt all the patients were able to completely excrete cobalt ions within 5-7 months after metal-on-metal bearing removal, for chromium ions it didn’t happen. If on the one hand the toxicokinetic differences between the two types of ions are confirmed by toxicological and occupational studies, on the other hand, this peculiar way of exposition represents a novel and important point of view. Thus, two different approaches were performed to better understand the subject specific capability to transport metal ions (albumin study) and to manage the response to them (heme-oxygenase-1 study): - a mutational screening of ALBUMIN gene was conducted in 30 MoM prosthetic patients resulting in the absence of nucleotidic changes compared with the ALB reference sequence. To this study was also added the analysis of expression of modified albumin protein; - a gene and protein expression study on 44 patients of heme-oxygenase-1, that is one of the most important antioxidant enzyme induced by metallic ions, was performed. This study resulted in no statistically significant differences in the expression of the gene and protein heme-oxygenase-1 between prosthetic and non-prosthetic patients, as well as between patients with high and low ions levels. Our results show that the protein studied (albumin and heme-oxygenase-1) seem to be not involved in determining chromium and cobalt ions level. On the other hand, achromium and cobalt elimination rates are different, but similar in all patients analyzed, suggesting that this process could be not patient-related. We support the importance of researching more about ions transport within the organism once released by hip prosthesis, about the chemical species involved, the districts where they are contained and the mechanisms of elimination, not excluding the existence of a subjective susceptibility to these metals ions.

Keywords: chromium, cobalt, hip prosthesis, individual susceptibility

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9 Epiphytic Growth on Filamentous Bacteria Found in Activated Sludge: A Morphological Approach

Authors: Thobela Conco, Sheena Kumari, Thor Stenstrom, Simona Rosetti, Valter Tandoi, Faizal Bux


Filamentous bacteria are well documented as causative agents of bulking and foaming in the biological wastewater treatment process. These filamentous bacteria are however closely associated with other non-filamentous organism forming a micro-niche. Among these specific epiphytic bacteria attach to filaments in the consortium of organisms that make up the floc. Neither the eco-physiological role of the epiphytes nor the nature of the interaction between the epiphytic bacteria and the filament hosts they colonize is well understood and in need of in-depth investigations. The focus of this presentation is on the interaction between the epiphytic bacteria and the filament host. Samples from the activated sludge treatment have been repeatedly collected from several wastewater treatment plants in KwaZulu Natal. Extensive investigations have been performed with SEM and TEM electron microscopy, Polarized Light Microscopy with Congo red staining, and Thioflavin T staining to document the interaction. SEM was used to document the morphology of both the filament host and their epiphytes counterparts with the focus on the interface/point of contact between the two, while the main focus of the TEM investigations with the higher magnification aimed to document the ultra-structure features of two organisms relating to the interaction. The interaction of the perpendicular attachment partly seems to be governed by the physiological status of the filaments. The attachment further seems to trigger a response in the filaments with distinct internal visible structures at the attachment sites. It is postulated that these structures most likely are amyloid fibrils. Amyloid fibrils may play an overarching role in different types of attachments and has earlier been noted to play a significant role in biofilm formation in activated sludge. They also play a medical role in degenerative diseases such as Alzheimer’s and Diabetes. Further studies aims to define the eco-physiological role of amyloid fibrils in filamentous bacteria, based on their observed presence at interaction sites in this study. This will also relate to additional findings where selectivity within the species of epiphytes attaching to the selected filaments has been noted. The practical implications of the research findings is still to be determined, but the ecophysiological interaction between two closely associated species or groups may have significant impact in the future understanding of wastewater treatment processes and broaden existing knowledge on population dynamics.

Keywords: activated sludge, amyloid proteins, epiphytic bacteria, filamentous bacteria

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8 Microfluidic Plasmonic Bio-Sensing of Exosomes by Using a Gold Nano-Island Platform

Authors: Srinivas Bathini, Duraichelvan Raju, Simona Badilescu, Muthukumaran Packirisamy


A bio-sensing method, based on the plasmonic property of gold nano-islands, has been developed for detection of exosomes in a clinical setting. The position of the gold plasmon band in the UV-Visible spectrum depends on the size and shape of gold nanoparticles as well as on the surrounding environment. By adsorbing various chemical entities, or binding them, the gold plasmon band will shift toward longer wavelengths and the shift is proportional to the concentration. Exosomes transport cargoes of molecules and genetic materials to proximal and distal cells. Presently, the standard method for their isolation and quantification from body fluids is by ultracentrifugation, not a practical method to be implemented in a clinical setting. Thus, a versatile and cutting-edge platform is required to selectively detect and isolate exosomes for further analysis at clinical level. The new sensing protocol, instead of antibodies, makes use of a specially synthesized polypeptide (Vn96), to capture and quantify the exosomes from different media, by binding the heat shock proteins from exosomes. The protocol has been established and optimized by using a glass substrate, in order to facilitate the next stage, namely the transfer of the protocol to a microfluidic environment. After each step of the protocol, the UV-Vis spectrum was recorded and the position of gold Localized Surface Plasmon Resonance (LSPR) band was measured. The sensing process was modelled, taking into account the characteristics of the nano-island structure, prepared by thermal convection and annealing. The optimal molar ratios of the most important chemical entities, involved in the detection of exosomes were calculated as well. Indeed, it was found that the results of the sensing process depend on the two major steps: the molar ratios of streptavidin to biotin-PEG-Vn96 and, the final step, the capture of exosomes by the biotin-PEG-Vn96 complex. The microfluidic device designed for sensing of exosomes consists of a glass substrate, sealed by a PDMS layer that contains the channel and a collecting chamber. In the device, the solutions of linker, cross-linker, etc., are pumped over the gold nano-islands and an Ocean Optics spectrometer is used to measure the position of the Au plasmon band at each step of the sensing. The experiments have shown that the shift of the Au LSPR band is proportional to the concentration of exosomes and, thereby, exosomes can be accurately quantified. An important advantage of the method is the ability to discriminate between exosomes having different origins.

Keywords: exosomes, gold nano-islands, microfluidics, plasmonic biosensing

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7 CSPG4 Molecular Target in Canine Melanoma, Osteosarcoma and Mammary Tumors for Novel Therapeutic Strategies

Authors: Paola Modesto, Floriana Fruscione, Isabella Martini, Simona Perga, Federica Riccardo, Mariateresa Camerino, Davide Giacobino, Cecilia Gola, Luca Licenziato, Elisabetta Razzuoli, Katia Varello, Lorella Maniscalco, Elena Bozzetta, Angelo Ferrari


Canine and human melanoma, osteosarcoma (OSA), and mammary carcinomas are aggressive tumors with common characteristics making dogs a good model for comparative oncology. Novel therapeutic strategies against these tumors could be useful to both species. In humans, chondroitin sulphate proteoglycan 4 (CSPG4) is a marker involved in tumor progression and could be a candidate target for immunotherapy. The anti-CSPG4 DNA electrovaccination has shown to be an effective approach for canine malignant melanoma (CMM) [1]. An immunohistochemistry evaluation of CSPG4 expression in tumour tissue is generally performed prior to electrovaccination. To assess the possibility to perform a rapid molecular evaluation and in order to validate these spontaneous canine tumors as the model for human studies, we investigate the CSPG4 gene expression by RT qPCR in CMM, OSA, and canine mammary tumors (CMT). The total RNA was extracted from RNAlater stored tissue samples (CMM n=16; OSA n=13; CMT n=6; five paired normal tissues for CMM, five paired normal tissues for OSA and one paired normal tissue for CMT), retro-transcribed and then analyzed by duplex RT-qPCR using two different TaqMan assays for the target gene CSPG4 and the internal reference gene (RG) Ribosomal Protein S19 (RPS19). RPS19 was selected from a panel of 9 candidate RGs, according to NormFinder analysis following the protocol already described [2]. Relative expression was analyzed by CFX Maestro™ Software. Student t-test and ANOVA were performed (significance set at P<0.05). Results showed that gene expression of CSPG4 in OSA tissues is significantly increased by 3-4 folds when compared to controls. In CMT, gene expression of the target was increased from 1.5 to 19.9 folds. In melanoma, although an increasing trend was observed, no significant differences between the two groups were highlighted. Immunohistochemistry analysis of the two cancer types showed that the expression of CSPG4 within CMM is concentrated in isles of cells compared to OSA, where the distribution of positive cells is homogeneous. This evidence could explain the differences in gene expression results.CSPG4 immunohistochemistry evaluation in mammary carcinoma is in progress. The evidence of CSPG4 expression in a different type of canine tumors opens the way to the possibility of extending the CSPG4 immunotherapy marker in CMM, OSA, and CMT and may have an impact to translate this strategy modality to human oncology.

Keywords: canine melanoma, canine mammary carcinomas, canine osteosarcoma, CSPG4, gene expression, immunotherapy

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6 Analysis of Differentially Expressed Genes in Spontaneously Occurring Canine Melanoma

Authors: Simona Perga, Chiara Beltramo, Floriana Fruscione, Isabella Martini, Federica Cavallo, Federica Riccardo, Paolo Buracco, Selina Iussich, Elisabetta Razzuoli, Katia Varello, Lorella Maniscalco, Elena Bozzetta, Angelo Ferrari, Paola Modesto


Introduction: Human and canine melanoma have common clinical, histologic characteristics making dogs a good model for comparative oncology. The identification of specific genes and a better understanding of the genetic landscape, signaling pathways, and tumor–microenvironmental interactions involved in the cancer onset and progression is essential for the development of therapeutic strategies against this tumor in both species. In the present study, the differential expression of genes in spontaneously occurring canine melanoma and in paired normal tissue was investigated by targeted RNAseq. Material and Methods: Total RNA was extracted from 17 canine malignant melanoma (CMM) samples and from five paired normal tissues stored in RNA-later. In order to capture the greater genetic variability, gene expression analysis was carried out using two panels (Qiagen): Human Immuno-Oncology (HIO) and Mouse-Immuno-Oncology (MIO) and the miSeq platform (Illumina). These kits allow the detection of the expression profile of 990 genes involved in the immune response against tumors in humans and mice. The data were analyzed through the CLCbio Genomics Workbench (Qiagen) software using the Canis lupus familiaris genome as a reference. Data analysis were carried out both comparing the biologic group (tumoral vs. healthy tissues) and comparing neoplastic tissue vs. paired healthy tissue; a Fold Change greater than two and a p-value less than 0.05 were set as the threshold to select interesting genes. Results and Discussion: Using HIO 63, down-regulated genes were detected; 13 of those were also down-regulated comparing neoplastic sample vs. paired healthy tissue. Eighteen genes were up-regulated, 14 of those were also down-regulated comparing neoplastic sample vs. paired healthy tissue. Using the MIO, 35 down regulated-genes were detected; only four of these were down-regulated, also comparing neoplastic sample vs. paired healthy tissue. Twelve genes were up-regulated in both types of analysis. Considering the two kits, the greatest variation in Fold Change was in up-regulated genes. Dogs displayed a greater genetic homology with humans than mice; moreover, the results have shown that the two kits are able to detect different genes. Most of these genes have specific cellular functions or belong to some enzymatic categories; some have already been described to be correlated to human melanoma and confirm the validity of the dog as a model for the study of molecular aspects of human melanoma.

Keywords: animal model, canine melanoma, gene expression, spontaneous tumors, targeted RNAseq

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5 Gold-Mediated Modification of Apoferritin Surface with Targeting Antibodies

Authors: Simona Dostalova, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek


Protein apoferritin seems to be a very promising structure for use as a nanocarrier. It is prepared from intracellular ferritin protein naturally found in most organisms. The role of ferritin proteins is to store and transport ferrous ions. Apoferritin is a hollow protein cage without ferrous ions that can be prepared from ferritin by reduction with thioglycolic acid or dithionite. The structure of apoferritin is composed of 24 protein subunits, creating a sphere with 12 nm in diameter. The inner cavity has a diameter of 8 nm. The drug encapsulation process is based on the response of apoferritin structure to the pH changes of surrounding solution. In low pH, apoferritin is disassembled into individual subunits and its structure is “opened”. It can then be mixed with any desired cytotoxic drug and after adjustment of pH back to neutral the subunits are reconnected again and the drug is encapsulated within the apoferritin particles. Excess drug molecules can be removed by dialysis. The receptors for apoferritin, SCARA5 and TfR1 can be found in the membrane of both healthy and cancer cells. To enhance the specific targeting of apoferritin nanocarrier, it is possible to modify its surface with targeting moieties, such as antibodies. To ensure sterically correct complex, we used a a peptide linker based on a protein G with N-terminus affinity towards Fc region of antibodies. To connect the peptide to the surface of apoferritin, the C-terminus of peptide was made of cysteine with affinity to gold. The surface of apoferritin with encapsulated doxorubicin (ApoDox) was coated either with gold nanoparticles (ApoDox-Nano) or gold (III) chloride hydrate reduced with sodium borohydride (ApoDox-HAu). The applied amount of gold in form of gold (III) chloride hydrate was 10 times higher than in the case of gold nanoparticles. However, after removal of the excess unbound ions by electrophoretic separation, the concentration of gold on the surface of apoferritin was only 6 times higher for ApoDox-HAu in comparison with ApoDox-Nano. Moreover, the reduction with sodium borohydride caused a loss of doxorubicin fluorescent properties (excitation maximum at 480 nm with emission maximum at 600 nm) and thus its biological activity. Fluorescent properties of ApoDox-Nano were similar to the unmodified ApoDox, therefore it was more suited for the intended use. To evaluate the specificity of apoferritin modified with antibodies, we used ELISA-like method with the surface of microtitration plate wells coated by the antigen (goat anti-human IgG antibodies). To these wells, we applied ApoDox without targeting antibodies and ApoDox-Nano modified with targeting antibodies (human IgG antibodies). The amount of unmodified ApoDox on antigen after incubation and subsequent rinsing with water was 5 times lower than in the case of ApoDox-Nano modified with targeting antibodies. The modification of non-gold ApoDox with antibodies caused no change in its targeting properties. It can therefore be concluded that the demonstrated procedure allows us to create nanocarrier with enhanced targeting properties, suitable for nanomedicine.

Keywords: apoferritin, doxorubicin, nanocarrier, targeting antibodies

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4 Prosodic Transfer in Foreign Language Learning: A Phonetic Crosscheck of Intonation and F₀ Range between Italian and German Native and Non-Native Speakers

Authors: Violetta Cataldo, Renata Savy, Simona Sbranna


Background: Foreign Language Learning (FLL) is characterised by prosodic transfer phenomena regarding pitch accents placement, intonation patterns, and pitch range excursion from the learners’ mother tongue to their Foreign Language (FL) which suggests that the gradual development of general linguistic competence in FL does not imply an equally correspondent improvement of the prosodic competence. Topic: The present study aims to monitor the development of prosodic competence of learners of Italian and German throughout the FLL process. The primary object of this study is to investigate the intonational features and the f₀ range excursion of Italian and German from a cross-linguistic perspective; analyses of native speakers’ productions point out the differences between this pair of languages and provide models for the Target Language (TL). A following crosscheck compares the L2 productions in Italian and German by non-native speakers to the Target Language models, in order to verify the occurrence of prosodic interference phenomena, i.e., type, degree, and modalities. Methodology: The subjects of the research are university students belonging to two groups: Italian native speakers learning German as FL and German native speakers learning Italian as FL. Both of them have been divided into three subgroups according to the FL proficiency level (beginners, intermediate, advanced). The dataset consists of wh-questions placed in situational contexts uttered in both speakers’ L1 and FL. Using a phonetic approach, analyses have considered three domains of intonational contours (Initial Profile, Nuclear Accent, and Terminal Contour) and two dimensions of the f₀ range parameter (span and level), which provide a basis for comparison between L1 and L2 productions. Findings: Results highlight a strong presence of prosodic transfer phenomena affecting L2 productions in the majority of both Italian and German learners, irrespective of their FL proficiency level; the transfer concerns all the three domains of the contour taken into account, although with different modalities and characteristics. Currently, L2 productions of German learners show a pitch span compression on the domain of the Terminal Contour compared to their L1 towards the TL; furthermore, German learners tend to use lower pitch range values in deviation from their L1 when improving their general linguistic competence in Italian FL proficiency level. Results regarding pitch range span and level in L2 productions by Italian learners are still in progress. At present, they show a similar tendency to expand the pitch span and to raise the pitch level, which also reveals a deviation from the L1 possibly in the direction of German TL. Conclusion: Intonational features seem to be 'resistant' parameters to which learners appear not to be particularly sensitive. By contrast, they show a certain sensitiveness to FL pitch range dimensions. Making clear which the most resistant and the most sensitive parameters are when learning FL prosody could lay groundwork for the development of prosodic trainings thanks to which learners could finally acquire a clear and natural pronunciation and intonation.

Keywords: foreign language learning, German, Italian, L2 prosody, pitch range, transfer

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3 Shocks and Flows - Employing a Difference-In-Difference Setup to Assess How Conflicts and Other Grievances Affect the Gender and Age Composition of Refugee Flows towards Europe

Authors: Christian Bruss, Simona Gamba, Davide Azzolini, Federico Podestà


In this paper, the authors assess the impact of different political and environmental shocks on the size and on the age and gender composition of asylum-related migration flows to Europe. With this paper, the authors contribute to the literature by looking at the impact of different political and environmental shocks on the gender and age composition of migration flows in addition to the size of these flows. Conflicting theories predict different outcomes concerning the relationship between political and environmental shocks and the migration flows composition. Analyzing the relationship between the causes of migration and the composition of migration flows could yield more insights into the mechanisms behind migration decisions. In addition, this research may contribute to better informing national authorities in charge of receiving these migrant, as women and children/the elderly require different assistance than young men. To be prepared to offer the correct services, the relevant institutions have to be aware of changes in composition based on the shock in question. The authors analyze the effect of different types of shocks on the number, the gender and age composition of first time asylum seekers originating from 154 sending countries. Among the political shocks, the authors consider: violence between combatants, violence against civilians, infringement of political rights and civil liberties, and state terror. Concerning environmental shocks, natural disasters (such as droughts, floods, epidemics, etc.) have been included. The data on asylum seekers applying to any of the 32 Schengen Area countries between 2008 and 2015 is on a monthly basis. Data on asylum applications come from Eurostat, data on shocks are retrieved from various sources: georeferenced conflict data come from the Uppsala Conflict Data Program (UCDP), data on natural disasters from the Centre for Research on the Epidemiology of Disasters (CRED), data on civil liberties and political rights from Freedom House, data on state terror from the Political Terror Scale (PTS), GDP and population data from the World Bank, and georeferenced population data from the Socioeconomic Data and Applications Center (SEDAC). The authors adopt a Difference-in-Differences identification strategy, exploiting the different timing of several kinds of shocks across countries. The highly skewed distribution of the dependent variable is taken into account by using count data models. In particular, a Zero Inflated Negative Binomial model is adopted. Preliminary results show that different shocks - such as armed conflict and epidemics - exert weak immediate effects on asylum-related migration flows and almost non-existent effects on the gender and age composition. However, this result is certainly affected by the fact that no time lags have been introduced so far. Finding the correct time lags depends on a great many variables not limited to distance alone. Therefore, finding the appropriate time lags is still a work in progress. Considering the ongoing refugee crisis, this topic is more important than ever. The authors hope that this research contributes to a less emotionally led debate.

Keywords: age, asylum, Europe, forced migration, gender

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2 Superparamagnetic Core Shell Catalysts for the Environmental Production of Fuels from Renewable Lignin

Authors: Cristina Opris, Bogdan Cojocaru, Madalina Tudorache, Simona M. Coman, Vasile I. Parvulescu, Camelia Bala, Bahir Duraki, Jeroen A. Van Bokhoven


The tremendous achievements in the development of the society concretized by more sophisticated materials and systems are merely based on non-renewable resources. Consequently, after more than two centuries of intensive development, among others, we are faced with the decrease of the fossil fuel reserves, an increased impact of the greenhouse gases on the environment, and economic effects caused by the fluctuations in oil and mineral resource prices. The use of biomass may solve part of these problems, and recent analyses demonstrated that from the perspective of the reduction of the emissions of carbon dioxide, its valorization may bring important advantages conditioned by the usage of genetic modified fast growing trees or wastes, as primary sources. In this context, the abundance and complex structure of lignin may offer various possibilities of exploitation. However, its transformation in fuels or chemicals supposes a complex chemistry involving the cleavage of C-O and C-C bonds and altering of the functional groups. Chemistry offered various solutions in this sense. However, despite the intense work, there are still many drawbacks limiting the industrial application. Thus, the proposed technologies considered mainly homogeneous catalysts meaning expensive noble metals based systems that are hard to be recovered at the end of the reaction. Also, the reactions were carried out in organic solvents that are not acceptable today from the environmental point of view. To avoid these problems, the concept of this work was to investigate the synthesis of superparamagnetic core shell catalysts for the fragmentation of lignin directly in the aqueous phase. The magnetic nanoparticles were covered with a nanoshell of an oxide (niobia) with a double role: to protect the magnetic nanoparticles and to generate a proper (acidic) catalytic function and, on this composite, cobalt nanoparticles were deposed in order to catalyze the C-C bond splitting. With this purpose, we developed a protocol to prepare multifunctional and magnetic separable nano-composite [email protected]@Fe3O4 catalysts. We have also established an analytic protocol for the identification and quantification of the fragments resulted from lignin depolymerization in both liquid and solid phase. The fragmentation of various lignins occurred on the prepared materials in high yields and with very good selectivity in the desired fragments. The optimization of the catalyst composition indicated a cobalt loading of 4wt% as optimal. Working at 180 oC and 10 atm H2 this catalyst allowed a conversion of lignin up to 60% leading to a mixture containing over 96% in C20-C28 and C29-C37 fragments that were then completely fragmented to C12-C16 in a second stage. The investigated catalysts were completely recyclable, and no leaching of the elements included in the composition was determined by inductively coupled plasma optical emission spectrometry (ICP-OES).

Keywords: superparamagnetic core-shell catalysts, environmental production of fuels, renewable lignin, recyclable catalysts

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