Search results for: Elena Elisabeta Manea
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 225

Search results for: Elena Elisabeta Manea

75 The Use of Food Industry Bio-Products for Sustainable Lactic Acid Bacteria Encapsulation

Authors: Paulina Zavistanaviciute, Vita Krungleviciute, Elena Bartkiene

Abstract:

Lactic acid bacteria (LAB) are microbial supplements that increase the nutritional, therapeutic, and safety value of food and feed. Often LAB strains are incubated in an expensive commercially available de Man-Rogosa-Sharpe (MRS) medium; the cultures are centrifuged, and the cells are washing with sterile water. Potato juice and apple juice industry bio-products are industrial wastes which may constitute a source of digestible nutrients for microorganisms. Due to their low cost and good chemical composition, potato juice and apple juice production bio- products could have a potential application in LAB encapsulation. In this study, pure LAB (P. acidilactici and P. pentosaceus) were multiplied in a crushed potato juice and apple juice industry bio-products medium. Before using, bio-products were sterilized and filtered. No additives were added to mass, except apple juice industry bioproducts were diluted with sterile water (1/5; v/v). The tap of sterilised mass, and LAB cell suspension (5 mL), containing of 8.9 log10 colony-forming units (cfu) per mL of the P. acidilactici and P. pentosaceus was used to multiply the LAB for 72 h. The final colony number in the potato juice and apple juice bio- products substrate was on average 9.60 log10 cfu/g. In order to stabilize the LAB, several methods of dehydration have been tested: lyophilisation (MilrockKieffer Lane, Kingston, USA) and dehydration in spray drying system (SD-06, Keison, Great Britain). Into the spray drying system multiplied LAB in a crushed potato juice and apple juice bio-products medium was injected in peristaltic way (inlet temperature +60 °C, inlet air temperature +150° C, outgoing air temperature +80 °C, air flow 200 m3/h). After lyophilisation (-48 °C) and spray drying (+150 °C) the viable cell concentration in the fermented potato juice powder was 9.18 ± 0.09 log10 cfu/g and 9.04 ± 0.07 log10 cfu/g, respectively, and in apple mass powder 8.03 ± 0.04 log10 cfu/g and 7.03 ± 0.03 log10 cfu/g, respectively. Results indicated that during the storage (after 12 months) at room temperature (22 +/- 2 ºC) LAB count in dehydrated products was 5.18 log10 cfu/g and 7.00 log10 cfu/g (in spray dried and lyophilized potato juice powder, respectively), and 3.05 log10 cfu/g and 4.10 log10 cfu/g (in spray dried and lyophilized apple juice industry bio-products powder, respectively). According to obtained results, potato juice could be used as alternative substrate for P. acidilactici and P. pentosaceus cultivation, and by drying received powders can be used in food/feed industry as the LAB starters. Therefore, apple juice industry by- products before spray drying and lyophilisation should be modified (i. e. by using different starches) in order to improve its encapsulation.

Keywords: bio-products, encapsulation, lactic acid bacteria, sustainability

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74 Wettability of Superhydrophobic Polymer Layers Filled with Hydrophobized Silica on Glass

Authors: Diana Rymuszka, Konrad Terpiłowski, Lucyna Hołysz, Elena Goncharuk, Iryna Sulym

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Superhydrophobic surfaces exhibit extremely high water repellency. The commonly accepted basic criterion for such surfaces is a water contact angle larger than 150°, low contact angle hysteresis and low sliding angle. These surfaces are of special interest, because properties such as anti-sticking, anti-contamination and self-cleaning are expected. These properties are attractive for many applications such as anti-sticking of snow for antennas and windows, anti-biofouling paints for boats, waterproof clothing, self-cleaning windshields for automobiles, dust-free coatings or metal refining. The various methods for the preparation of superhydrophobic surfaces since last two decades have been reported, such as phase separation, electrochemical deposition, template method, plasma method, chemical vapor deposition, wet chemical reaction, sol-gel processing, lithography and so on. The aim of the study was to investigate the influence of modified colloidal silica, used as a filler, on the hydrophobicity of the polymer film deposited on the glass support activated with plasma. On prepared surfaces water advancing (ӨA) and receding (ӨR) contact angles were measured and then their total apparent surface free energy was determined using the contact angle hysteresis approach (CAH). The structures of deposited films were observed with the help of an optical microscope. Topographies of selected films were also determined using an optical profilometer. It was found that plasma treatment influence glass surface wetting and energetic properties that is observed in higher adhesion between polymer/filler film and glass support. Using the colloidal silica particles as a filler for the polymer thin film deposited on the glass support, it is possible to produce strongly adhering layers of superhydrophobic properties. The best superhydrophobic properties were obtained for surfaces of the film glass/polimer + modified silica covered in 89 and 100%. The advancing contact angle measured on these surfaces amounts above 150° that leads to under 2 mJ/m2 value of the apparent surface free energy. Such films may have many practical applications, among others, as dust-free coatings or anticorrosion protection.

Keywords: contact angle, plasma, superhydrophobic, surface free energy

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73 ESDN Expression in the Tumor Microenvironment Coordinates Melanoma Progression

Authors: Roberto Coppo, Francesca Orso, Daniela Dettori, Elena Quaglino, Lei Nie, Mehran M. Sadeghi, Daniela Taverna

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Malignant melanoma is currently the fifth most common cancer in the white population and it is fatal in its metastatic stage. Several research studies in recent years have provided evidence that cancer initiation and progression are driven by genetic alterations of the tumor and paracrine interactions between tumor and microenvironment. Scattered data show that the Endothelial and Smooth muscle cell-Derived Neuropilin-like molecule (ESDN) controls cell proliferation and movement of stroma and tumor cells. To investigate the role of ESDN in the tumor microenvironment during melanoma progression, murine melanoma cells (B16 or B16-F10) were injected in ESDN knockout mice in order to evaluate how the absence of ESDN in stromal cells could influence melanoma progression. While no effect was found on primary tumor growth, increased cell extravasation and lung metastasis formation was observed in ESDN knockout mice compared to wild type controls. In order to understand how cancer cells cross the endothelial barrier during metastatic dissemination in an ESDN-null microenvironment, structure, and permeability of lung blood vessels were analyzed. Interestingly, ESDN knockout mice showed structurally altered and more permeable vessels compared to wild type animals. Since cell surface molecules mediate the process of tumor cell extravasation, the expression of a panel of extravasation-related ligands and receptors was analyzed. Importantly, modulations of N-cadherin, E-selectin, ICAM-1 and VAP-1 were observed in ESDN knockout endothelial cells, suggesting the presence of a favorable tumor microenvironment which facilitates melanoma cell extravasation and metastasis formation in the absence of ESDN. Furthermore, a potential contribution of immune cells in tumor dissemination was investigated. An increased recruitment of macrophages in the lungs of ESDN knockout mice carrying subcutaneous B16-F10 tumors was found. In conclusion, our data suggest a functional role of ESDN in the tumor microenvironment during melanoma progression and the identification of the mechanisms that regulate tumor cell extravasation could lead to the development of new therapies to reduce metastasis formation.

Keywords: melanoma, tumor microenvironment, extravasation, cell surface molecules

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72 The Impact of Trait and Mathematical Anxiety on Oscillatory Brain Activity during Lexical and Numerical Error-Recognition Tasks

Authors: Alexander N. Savostyanov, Tatyana A. Dolgorukova, Elena A. Esipenko, Mikhail S. Zaleshin, Margherita Malanchini, Anna V. Budakova, Alexander E. Saprygin, Yulia V. Kovas

Abstract:

The present study compared spectral-power indexes and cortical topography of brain activity in a sample characterized by different levels of trait and mathematical anxiety. 52 healthy Russian-speakers (age 17-32; 30 males) participated in the study. Participants solved an error recognition task under 3 conditions: A lexical condition (simple sentences in Russian), and two numerical conditions (simple arithmetic and complicated algebraic problems). Trait and mathematical anxiety were measured using self-repot questionnaires. EEG activity was recorded simultaneously during task execution. Event-related spectral perturbations (ERSP) were used to analyze spectral-power changes in brain activity. Additionally, sLORETA was applied in order to localize the sources of brain activity. When exploring EEG activity recorded after tasks onset during lexical conditions, sLORETA revealed increased activation in frontal and left temporal cortical areas, mainly in the alpha/beta frequency ranges. When examining the EEG activity recorded after task onset during arithmetic and algebraic conditions, additional activation in delta/theta band in the right parietal cortex was observed. The ERSP plots reveled alpha/beta desynchronizations within a 500-3000 ms interval after task onset and slow-wave synchronization within an interval of 150-350 ms. Amplitudes of these intervals reflected the accuracy of error recognition, and were differently associated with the three (lexical, arithmetic and algebraic) conditions. The level of trait anxiety was positively correlated with the amplitude of alpha/beta desynchronization. The level of mathematical anxiety was negatively correlated with the amplitude of theta synchronization and of alpha/beta desynchronization. Overall, trait anxiety was related with an increase in brain activation during task execution, whereas mathematical anxiety was associated with increased inhibitory-related activity. We gratefully acknowledge the support from the №11.G34.31.0043 grant from the Government of the Russian Federation.

Keywords: anxiety, EEG, lexical and numerical error-recognition tasks, alpha/beta desynchronization

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71 Posterior Acetabular Fractures-Optimizing the Treatment by Enhancing Practical Skills

Authors: Olivera Lupescu, Taina Elena Avramescu, Mihail Nagea, Alexandru Dimitriu

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Acetabular fractures represent a real challenge due to their impact upon the long term function of the hip joint, and due to the risk of intra- and peri-operative complications especially that they affect young, active people. That is why treating these fractures require certain skills which must be exercised, regarding the pre-operative planning, as well as the execution of surgery.The authors retrospectively analyse 38 cases with acetabular fractures operated using the posterior approach in our hospital between 01.01.2013- 01.01.2015 for which complete medical records ensure a follow-up of 24 months, in order to establish the main causes of potential errors and to underline the methods for preventing them. This target is included in the Erasmus + project ‘Collaborative learning for enhancing practical skills for patient-focused interventions in gait rehabilitation after orthopedic surgery COR-skills’. This paper analyses the pitfalls revealed by these cases, as well as the measures necessary to enhance the practical skills of the surgeons who perform acetabular surgery. Pre-op planning matched the intra and post-operative outcome in 88% of the analyzed points, from 72% at the beginning to 94% in the last case, meaning that experience is very important in treating this injury. The main problems detected for the posterior approach were: nervous complications - 3 cases, 1 of them a complete paralysis of the sciatic nerve, which recovered 6 months after surgery, and in other 2 cases intra-articular position of the screws was demonstrated by post-operative CT scans, so secondary screw removal was necessary in these cases. We analysed this incident, too, due to lack of information about the relationship between the screws and the joint secondary to this approach. Septic complications appeared in 3 cases, 2 superficial and 1 profound (requiring implant removal). The most important problems were the reduction of the fractures and the positioning of the screws so as not to interfere with the the articular space. In posterior acetabular fractures, pre-op complex planning is important in order to achieve maximum treatment efficacy with minimum of risk; an optimal training of the surgeons insisting on the main points of potential mistakes ensure the success of the procedure, as well as a favorable outcome for the patient.

Keywords: acetabular fractures, articular congruency, surgical skills, vocational training

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70 Wastewater Treatment in the Abrasives Industry via Fenton and Photo-Fenton Oxidation Processes: A Case Study from Peru

Authors: Hernan Arturo Blas López, Gustavo Henndel Lopes, Antonio Carlos Silva Costa Teixeira, Carmen Elena Flores Barreda, Patricia Araujo Pantoja

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Phenols are toxic for life and the environment and may come from many sources. Uncured phenolic monomers present in phenolic resins used as binders in grinding wheels and emery paper can contaminate industrial wastewaters in abrasives manufacture plants. Furthermore, vestiges of resol and novolacs resins generated by wear and tear of abrasives are also possible sources of water contamination by phenolics in these facilities. Fortunately, advanced oxidation by dark Fenton and photo-Fenton techniques are capable of oxidizing phenols and their degradation products up to their mineralization into H₂O and CO₂. The maximal allowable concentrations for phenols in Peruvian waterbodies is very low, such that insufficiently treated effluents from the abrasives industry are a potential environmental noncompliance. The current case study highlights findings obtained during the lab-scale application of Fenton’s and photo-assisted Fenton’s chemistries to real industrial wastewater samples from an abrasives manufacture plant in Peru. The goal was to reduce the phenolic content and sample toxicity. For this purpose, two independent variables-reaction time and effect of ultraviolet radiation–were studied as for their impacts on the concentration of total phenols, total organic carbon (TOC), biological oxygen demand (BOD) and chemical oxygen demand (COD). In this study, diluted samples (1 L) of the industrial effluent were treated with Fenton’s reagent (H₂O₂ and Fe²⁺ from FeSO₄.H₂O) during 10 min in a photochemical batch reactor (Alphatec RFS-500, Brazil) at pH 2.92. In the case of photo-Fenton tests with ultraviolet lamps of 9 W, UV-A, UV-B and UV-C lamps were evaluated. All process conditions achieved 100% of phenols degraded within 5 minutes. TOC, BOD and COD decreased by 49%, 52% and 86% respectively (all processes together). However, Fenton treatment was not capable of reducing BOD, COD and TOC below a certain value even after 10 minutes, contrarily to photo-Fenton. It was also possible to conclude that the processes here studied degrade other compounds in addition to phenols, what is an advantage. In all cases, elevated effluent dilution factors and high amounts of oxidant agent impact negatively the overall economy of the processes here investigated.

Keywords: fenton oxidation, wastewater treatment, phenols, abrasives industry

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69 Water Balance in the Forest Basins Essential for the Water Supply in Central America

Authors: Elena Listo Ubeda, Miguel Marchamalo Sacristan

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The demand for water doubles every twenty years, at a rate which is twice as fast as the world´s population growth. Despite it´s great importance, water is one of the most degraded natural resources in the world, mainly because of the reduction of natural vegetation coverage, population growth, contamination and changes in the soil use which reduces its capacity to collect water. This situation is especially serious in Central America, as reflected in the Human Development reports. The objective of this project is to assist in the improvement of water production and quality in Central America. In order to do these two watersheds in Costa Rica were selected as experiments: that of the Virilla-Durazno River, located in the extreme north east of the central valley which has an Atlantic influence; and that of the Jabillo River, which flows directly into the Pacific. The Virilla river watershed is located over andisols, and that of the Jabillo River is over alfisols, and both are of great importance for water supply to the Greater Metropolitan Area and the future tourist resorts respectively, as well as for the production of agriculture, livestock and hydroelectricity. The hydrological reaction in different soil-cover complexes, varying from the secondary forest to natural vegetation and degraded pasture, was analyzed according to the evaluation of the properties of the soil, infiltration, soil compaction, as well as the effects of the soil cover complex on erosion, calculated by the C factor of the Revised Universal Soil Loss Equation (RUSLE). A water balance was defined for each watershed, in which the volume of water that enters and leaves were estimated, as well as the evapotranspiration, runoff, and infiltration. Two future scenarios, representing the implementation of reforestation and deforestation plans, were proposed, and were analyzed for the effects of the soil cover complex on the water balance in each case. The results obtained show an increase of the ground water recharge in the humid forest areas, and an extension of the study of the dry areas is proposed since the ground water recharge here is diminishing. These results are of great significance for the planning, design of Payment Schemes for Environmental Services and the improvement of the existing water supply systems. In Central America spatial planning is a priority, as are the watersheds, in order to assess the water resource socially and economically, and securing its availability for the future.

Keywords: Costa Rica, infiltration, soil, water

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68 Global Modeling of Drill String Dragging and Buckling in 3D Curvilinear Bore-Holes

Authors: Valery Gulyayev, Sergey Glazunov, Elena Andrusenko, Nataliya Shlyun

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Enhancement of technology and techniques for drilling deep directed oil and gas bore-wells are of essential industrial significance because these wells make it possible to increase their productivity and output. Generally, they are used for drilling in hard and shale formations, that is why their drivage processes are followed by the emergency and failure effects. As is corroborated by practice, the principal drilling drawback occurring in drivage of long curvilinear bore-wells is conditioned by the need to obviate essential force hindrances caused by simultaneous action of the gravity, contact and friction forces. Primarily, these forces depend on the type of the technological regime, drill string stiffness, bore-hole tortuosity and its length. They can lead to the Eulerian buckling of the drill string and its sticking. To predict and exclude these states, special mathematic models and methods of computer simulation should play a dominant role. At the same time, one might note that these mechanical phenomena are very complex and only simplified approaches (‘soft string drag and torque models’) are used for their analysis. Taking into consideration that now the cost of directed wells increases essentially with complication of their geometry and enlargement of their lengths, it can be concluded that the price of mistakes of the drill string behavior simulation through the use of simplified approaches can be very high and so the problem of correct software elaboration is very urgent. This paper deals with the problem of simulating the regimes of drilling deep curvilinear bore-wells with prescribed imperfect geometrical trajectories of their axial lines. On the basis of the theory of curvilinear flexible elastic rods, methods of differential geometry, and numerical analysis methods, the 3D ‘stiff-string drag and torque model’ of the drill string bending and the appropriate software are elaborated for the simulation of the tripping in and out regimes and drilling operations. It is shown by the computer calculations that the contact and friction forces can be calculated and regulated, providing predesigned trouble-free modes of operation. The elaborated mathematic models and software can be used for the emergency situations prognostication and their exclusion at the stages of the drilling process design and realization.

Keywords: curvilinear drilling, drill string tripping in and out, contact forces, resistance forces

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67 Issues of Accounting of Lease and Revenue according to International Financial Reporting Standards

Authors: Nadezhda Kvatashidze, Elena Kharabadze

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It is broadly known that lease is a flexible means of funding enterprises. Lease reduces the risk related to access and possession of assets, as well as obtainment of funding. Therefore, it is important to refine lease accounting. The lease accounting regulations under the applicable standard (International Accounting Standards 17) make concealment of liabilities possible. As a result, the information users get inaccurate and incomprehensive information and have to resort to an additional assessment of the off-balance sheet lease liabilities. In order to address the problem, the International Financial Reporting Standards Board decided to change the approach to lease accounting. With the deficiencies of the applicable standard taken into account, the new standard (IFRS 16 ‘Leases’) aims at supplying appropriate and fair lease-related information to the users. Save certain exclusions; the lessee is obliged to recognize all the lease agreements in its financial report. The approach was determined by the fact that under the lease agreement, rights and obligations arise by way of assets and liabilities. Immediately upon conclusion of the lease agreement, the lessee takes an asset into its disposal and assumes the obligation to effect the lease-related payments in order to meet the recognition criteria defined by the Conceptual Framework for Financial Reporting. The payments are to be entered into the financial report. The new lease accounting standard secures supply of quality and comparable information to the financial information users. The International Accounting Standards Board and the US Financial Accounting Standards Board jointly developed IFRS 15: ‘Revenue from Contracts with Customers’. The standard allows the establishment of detailed revenue recognition practical criteria such as identification of the performance obligations in the contract, determination of the transaction price and its components, especially price variable considerations and other important components, as well as passage of control over the asset to the customer. IFRS 15: ‘Revenue from Contracts with Customers’ is very similar to the relevant US standards and includes requirements more specific and consistent than those of the standards in place. The new standard is going to change the recognition terms and techniques in the industries, such as construction, telecommunications (mobile and cable networks), licensing (media, science, franchising), real property, software etc.

Keywords: assessment of the lease assets and liabilities, contractual liability, division of contract, identification of contracts, contract price, lease identification, lease liabilities, off-balance sheet, transaction value

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66 Building up Regional Innovation Systems (RIS) for Development: The Case Study of the State of Mexico, México

Authors: Jose Luis Solleiro, Rosario Castanon, Laura Elena Martinez

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The State of Mexico is an administrative entity of Mexico, and it is one of the most important territories due to its great economic and social impact for the whole country, especially since it contributes with more than eight of the national Gross Domestic Product (GDP). The State of Mexico has a population of over seventeen million people and host very important business and productive industries such as Automotive, Chemicals, Pharmaceutical, and Agri-food. In 2017, the State Development Plan (Plan Estatal de Desarrollo in Spanish) which is a policy document that rules State's economic actions and integrates the bases for sectoral and regional programs to achieve regional development), raised innovation as a key aspect to boost competitiveness and productivity of the State of Mexico. Therefore, in line with this proposal, in 2018 the Mexican Council for Science and Technology (COMECYT for its acronym in Spanish), an institution in charge of promoting public science and technology policies in the State of Mexico, took actions towards building up the State´s Innovation System. Hence, the main objective of this paper is to review and analyze the process to create RIS in the State of Mexico. We focus on the key elements of the process, the diverse actors that were involved in it, the activities that were carried out and the identification of the challenges, findings, successes, and failures of the intended exercise. The methodology used to analyze the structure of the Innovation System of the State of Mexico is based on two elements: the case study and the research-action approach. The main objective of the paper, the case study was based on semi-structured interviews with key actors who have participated in the process of launching the RIS of the State of Mexico. Additionally, we analyzed the information reports and other documents that were elaborated during the process of shaping the State's innovation system. Finally, the results obtained in the process were also examined. The relevance of this investigation fundamentally rests in two elements: 1) keeping documental record of the process of building a RIS in Mexico; and 2) carrying out the analysis of this case study recognizing the importance of knowledge extraction and dissemination, so that lessons on this matter may be useful for similar experiences in the future. We conclude that in Mexico, documentation and analysis efforts related to the formation of RIS and interaction processes between innovation ecosystem actors are scarce, so documents like are of great importance, especially since it generates a series of findings and recommendations for the building of RIS.

Keywords: regional innovation systems, innovation, development, competitiveness

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65 Soils Properties of Alfisols in the Nicoya Peninsula, Guanacaste, Costa Rica

Authors: Elena Listo, Miguel Marchamalo

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This research studies the soil properties located in the watershed of Jabillo River in the Guanacaste province, Costa Rica. The soils are classified as Alfisols (T. Haplustalfs), in the flatter parts with grazing as Fluventic Haplustalfs or as a consequence of bad drainage as F. Epiaqualfs. The objective of this project is to define the status of the soil, to use remote sensing as a tool for analyzing the evolution of land use and determining the water balance of the watershed in order to improve the efficiency of the water collecting systems. Soil samples were analyzed from trial pits taken from secondary forests, degraded pastures, mature teak plantation, and regrowth -Tectona grandis L. F.- species developed favorably in the area. Furthermore, to complete the study, infiltration measurements were taken with an artificial rainfall simulator, as well as studies of soil compaction with a penetrometer, in points strategically selected from the different land uses. Regarding remote sensing, nearly 40 data samples were collected per plot of land. The source of radiation is reflected sunlight from the beam and the underside of leaves, bare soil, streams, roads and logs, and soil samples. Infiltration reached high levels. The majority of data came from the secondary forest and mature planting due to a high proportion of organic matter, relatively low bulk density, and high hydraulic conductivity. Teak regrowth had a low rate of infiltration because the studies made regarding the soil compaction showed a partial compaction over 50 cm. The secondary forest presented a compaction layer from 15 cm to 30 cm deep, and the degraded pasture, as a result of grazing, in the first 15 cm. In this area, the alfisols soils have high content of iron oxides, a fact that causes a higher reflectivity close to the infrared region of the electromagnetic spectrum (around 700mm), as a result of clay texture. Specifically in the teak plantation where the reflectivity reaches values of 90 %, this is due to the high content of clay in relation to others. In conclusion, the protective function of secondary forests is reaffirmed with regards to erosion and high rate of infiltration. In humid climates and permeable soils, the decrease of runoff is less, however, the percolation increases. The remote sensing indicates that being clay soils, they retain moisture in a better way and it means a low reflectivity despite being fine texture.

Keywords: alfisols, Costa Rica, infiltration, remote sensing

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64 Long Term Survival after a First Transient Ischemic Attack in England: A Case-Control Study

Authors: Padma Chutoo, Elena Kulinskaya, Ilyas Bakbergenuly, Nicholas Steel, Dmitri Pchejetski

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Transient ischaemic attacks (TIAs) are warning signs for future strokes. TIA patients are at increased risk of stroke and cardio-vascular events after a first episode. A majority of studies on TIA focused on the occurrence of these ancillary events after a TIA. Long-term mortality after TIA received only limited attention. We undertook this study to determine the long-term hazards of all-cause mortality following a first episode of a TIA using anonymised electronic health records (EHRs). We used a retrospective case-control study using electronic primary health care records from The Health Improvement Network (THIN) database. Patients born prior to or in year 1960, resident in England, with a first diagnosis of TIA between January 1986 and January 2017 were matched to three controls on age, sex and general medical practice. The primary outcome was all-cause mortality. The hazards of all-cause mortality were estimated using a time-varying Weibull-Cox survival model which included both scale and shape effects and a random frailty effect of GP practice. 20,633 cases and 58,634 controls were included. Cases aged 39 to 60 years at the first TIA event had the highest hazard ratio (HR) of mortality compared to matched controls (HR = 3.04, 95% CI (2.91 - 3.18)). The HRs for cases aged 61-70 years, 71-76 years and 77+ years were 1.98 (1.55 - 2.30), 1.79 (1.20 - 2.07) and 1.52 (1.15 - 1.97) compared to matched controls. Aspirin provided long-term survival benefits to cases. Cases aged 39-60 years on aspirin had HR of 0.93 (0.84 - 1.00), 0.90 (0.82 - 0.98) and 0.88 (0.80 - 0.96) at 5 years, 10 years and 15 years, respectively, compared to cases in the same age group who were not on antiplatelets. Similar beneficial effects of aspirin were observed in other age groups. There were no significant survival benefits with other antiplatelet options. No survival benefits of antiplatelet drugs were observed in controls. Our study highlights the excess long-term risk of death of TIA patients and cautions that TIA should not be treated as a benign condition. The study further recommends aspirin as the better option for secondary prevention for TIA patients compared to clopidogrel recommended by NICE guidelines. Management of risk factors and treatment strategies should be important challenges to reduce the burden of disease.

Keywords: dual antiplatelet therapy (DAPT), General Practice, Multiple Imputation, The Health Improvement Network(THIN), hazard ratio (HR), Weibull-Cox model

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63 Analyzing Water Waves in Underground Pumped Storage Reservoirs: A Combined 3D Numerical and Experimental Approach

Authors: Elena Pummer, Holger Schuettrumpf

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By today underground pumped storage plants as an outstanding alternative for classical pumped storage plants do not exist. They are needed to ensure the required balance between production and demand of energy. As a short to medium term storage pumped storage plants have been used economically over a long period of time, but their expansion is limited locally. The reasons are in particular the required topography and the extensive human land use. Through the use of underground reservoirs instead of surface lakes expansion options could be increased. Fulfilling the same functions, several hydrodynamic processes result in the specific design of the underground reservoirs and must be implemented in the planning process of such systems. A combined 3D numerical and experimental approach leads to currently unknown results about the occurring wave types and their behavior in dependence of different design and operating criteria. For the 3D numerical simulations, OpenFOAM was used and combined with an experimental approach in the laboratory of the Institute of Hydraulic Engineering and Water Resources Management at RWTH Aachen University, Germany. Using the finite-volume method and an explicit time discretization, a RANS-Simulation (k-ε) has been run. Convergence analyses for different time discretization, different meshes etc. and clear comparisons between both approaches lead to the result, that the numerical and experimental models can be combined and used as hybrid model. Undular bores partly with secondary waves and breaking bores occurred in the underground reservoir. Different water levels and discharges change the global effects, defined as the time-dependent average of the water level as well as the local processes, defined as the single, local hydrodynamic processes (water waves). Design criteria, like branches, directional changes, changes in cross-section or bottom slope, as well as changes in roughness have a great effect on the local processes, the global effects remain unaffected. Design calculations for underground pumped storage plants were developed on the basis of existing formulae and the results of the hybrid approach. Using the design calculations reservoirs heights as well as oscillation periods can be determined and lead to the knowledge of construction and operation possibilities of the plants. Consequently, future plants can be hydraulically optimized applying the design calculations on the local boundary conditions.

Keywords: energy storage, experimental approach, hybrid approach, undular and breaking Bores, 3D numerical approach

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62 Pelvic Floor Training in Elite Athletes: Fact or Fiction

Authors: Maria Barbano Acevedo-Gomez, Elena Sonsoles Rodriguez-Lopez, Sofia Olivia Calvo-Moreno, Angel Basas-Garcia, Cristophe Ramirez

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Introduction: Urinary incontinence (UI) is defined as the involuntary leakage of urine. In persons who practice sport, its prevalence is 36.1% (95% CI 26.5%-46.8%) and varies as it seems to depend on the intensity of exercise, movements, and impact on the ground. Such high impact sports are likely to generate higher intra-abdominal pressures and leading to pelvic floor muscle weakness. Even though the emphasis of this research is on female athletes, all women should perform pelvic floor muscle exercises as a part of their general physical exercise. Pelvic floor exercises are generally considered the first treatment against urinary incontinence. Objective: The main objective of the present study was to determine the knowledge of the pelvic floor and of the UI in elite athletes and know if they incorporate pelvic floor strengthening in their training. Methods: This was an observational study conducted on 754 elite athletes. After collecting questions about the pelvic floor, UI, and sport-related data, participants completed the questionnaire International Consultation on Incontinence Questionnaire-UI Short-Form (ICIQ-SF). Results: 57.3% of the athletes reflect not having knowledge of their pelvic floor, 48.3% do not know what strengthening exercises are, and around 90% have never practiced them. 78.1% (n=589) of all elite athletes do not include pelvic floor exercises in their training. Of the elite athletes surveyed, 33% had UI according to ICIQ-SF (mean age 23.75 ± 7.74 years). In response to the question 'Do you think you have or have had UI?', Only 9% of the 754 elite athletes admitted they presently had UI, and 13.3% indicated they had had UI at some time. However, 22.7% (n=171) reported they had experienced urine leakage while training. Of the athletes who indicated they did not have UI in the ICIQ-SF, 25.7% stated they did experience urine leakage during training (χ² [1] = 265.56; p < 0.001). Further, 12.3% of the athletes who considered they did not have UI and 60% of those who admitted they had had UI on some occasion stated they had suffered some urine leakage in the past 3 months (χ² [1] = 287.59; p < 0.001). Conclusions: There is a lack of knowledge about UI in sport. Through the use of validated questionnaires, we observed a UI prevalence of 33%, and 22.7% reported they experienced urine leakage while training. These figures contrast with only 9% of athletes who reported they had or had in the past had UI. This discrepancy could reflect the great lack of knowledge about UI in sports and that sometimes an athlete may consider that urine leakage is normal and a consequence of the demands of training. These data support the idea that coaches, physiotherapists, and other professionals involved in maximizing the performance of athletes should include pelvic floor muscle exercises in their training programs. Measures such as this could help to prevent UI during training and could be a starting point for future studies designed to develop adequate prevention and treatment strategies for this embarrassing problem affecting young athletes, both male and female.

Keywords: athletes, pelvic floor, performance, prevalence, sport, training, urinary incontinence

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61 TiO₂ Nanoparticles Induce DNA Damage and Expression of Biomarker of Oxidative Stress on Human Spermatozoa

Authors: Elena Maria Scalisi

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The increasing production and the use of TiO₂ nanoparticles (NPs) have inevitably led to their release into the environment, thereby posing a threat to organisms and also for human. Human exposure to TiO₂-NPs may occur during both manufacturing and use. TiO₂-NPs are common in consumer products for dermal application, toothpaste, food colorants, and nutritional supplements, then oral exposure may occur during use of such products. Into the body, TiO₂-NPs thanks to their small size (<100 nm), can, through testicular blood barrier inducing effect on testis and then on male reproductive health. The nanoscale size of TiO₂ increase the surface-to-volume ratio making them more reactive in a cell, then TiO₂ NPs increase their ability to produce reactive oxygen species (ROS). In male germ cells, ROS may have important implications in maintaining the normal functions of mature spermatozoa at physiological levels, moreover, in spermatozoa they are important signaling molecules for their hyperactivation and acrosome reaction. Nevertheless, an excess of ROS by external inputs such as NPs can increased the oxidative stress (OS), which results in damage DNA and apoptosis. The aim of our study has been investigate the impact of TiO₂ NPs on human spermatozoa, evaluating DNA damage and the expression of proteins involved in cell stress. According WHO guidelines 2021, we have exposed human spermatozoa in vitro to TiO₂ NP at concentrations 50 ppm, 100 ppm, 250 ppm, and 500 ppm for 1 hour (at 37°C and CO₂ at 5%). DNA damage was evaluated by Sperm Chromatin Dispersion Test (SCD) and TUNEL assay; moreover, we have evaluated the expression of biomarkers of oxidative stress like Heat Shock Protein 70 (HSP70) and Metallothioneins (MTs). Also, sperm parameters as motility viability have been evaluated. Our results not report a significant reduction in motility of spermatozoa at the end of the exposure. On the contrary, the progressive motility was increased at the highest concentration (500 ppm) and was statistically significant compared to control (p <0.05). Also, viability was not changed by exposure to TiO₂-NPs (p <0.05). However, increased DNA damage was observed at all concentrations, and the TUNEL assay highlighted the presence of single strand breaks in the DNA. The spermatozoa responded to the presence of TiO₂-NPs with the expression of Hsp70, which have a protective function because they allow the maintenance of cellular homeostasis in stressful/ lethal conditions. A positivity for MTs was observed mainly for the concentration of 4 mg/L. Although the biological and physiological function of the metallothionein (MTs) in the male genital organs is unclear, our results highlighted that the MTs expressed by spermatozoa maintain their biological role of detoxification from metals. Our results can give additional information to the data in the literature on the toxicity of TiO₂-NPs and reproduction.

Keywords: human spermatozoa, DNA damage, TiO₂-NPs, biomarkers

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60 Impact of the Dog-Technic for D1-D4 and Longitudinal Stroke Technique for Diaphragm on Peak Expiratory Flow (PEF) in Asthmatic Patients

Authors: Victoria Eugenia Garnacho-Garnacho, Elena Sonsoles Rodriguez-Lopez, Raquel Delgado-Delgado, Alvaro Otero-Campos, Jesus Guodemar-Perez, Angelo Michelle Vagali, Juan Pablo Hervas-Perez

Abstract:

Asthma is a heterogeneous disease which has always had a drug treatment. Osteopathic treatment that we propose is aimed, seen through a dorsal manipulation (Dog Technic D1-D4) and a technique for diaphragm (Longitudinal Stroke) forced expiratory flow in spirometry changes there are in particular that there is an increase in the volumes of the Peak Flow and Post intervention and effort and that the application of these two techniques together is more powerful if we applied only a Longitudinal (Stroke). Also rating if this type of treatment will have repercussions on breathlessness, a very common symptom in asthma. And finally to investigate if provided vertebra pain decreased after a manipulation. Methods—Participants were recruited between students and professors of the University, aged 18-65, patients (n = 18) were assigned randomly to one of the two groups, group 1 (longitudinal Stroke and manipulation dorsal Dog Technic) and group 2 (diaphragmatic technique, Longitudinal Stroke). The statistical analysis is characterized by the comparison of the main indicator of obstruction of via area PEF (peak expiratory flow) in various situations through the peak flow meter Datospir Peak-10. The measurements were carried out in four phases: at rest, after the stress test, after the treatment, after treatment and the stress test. After each stress test was evaluated, through the Borg scale, the level of Dyspnea on each patient, regardless of the group. In Group 1 in addition to these parameters was calculated using an algometer spinous pain before and after the manipulation. All data were taken at the minute. Results—12 Group 1 (Dog Technic and Longitudinal Stroke) patients responded positively to treatment, there was an increase of 5.1% and 6.1% of the post-treatment PEF and post-treatment, and effort. The results of the scale of Borg by which we measure the level of Dyspnea were positive, a 54.95%, patients noted an improvement in breathing. In addition was confirmed through the means of both groups group 1 in which two techniques were applied was 34.05% more effective than group 2 in which applied only a. After handling pain fell by 38% of the cases. Conclusions—The impact of the technique of Dog-Technic for D1-D4 and the Longitudinal Stroke technique for diaphragm in the volumes of peak expiratory flow (PEF) in asthmatic patients were positive, there was a change of the PEF Post intervention and post-treatment, and effort and showed the most effective group in which only a technique was applied. Furthermore this type of treatment decreased facilitated vertebrae pain and was efficient in the improvement of Dyspnea and the general well-being of the patient.

Keywords: ANS, asthma, manipulation, manual therapy, osteopathic

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59 Consumption and Diffusion Based Model of Tissue Organoid Development

Authors: Elena Petersen, Inna Kornienko, Svetlana Guryeva, Sergey Simakov

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In vitro organoid cultivation requires the simultaneous provision of necessary vascularization and nutrients perfusion of cells during organoid development. However, many aspects of this problem are still unsolved. The functionality of vascular network intergrowth is limited during early stages of organoid development since a function of the vascular network initiated on final stages of in vitro organoid cultivation. Therefore, a microchannel network should be created in early stages of organoid cultivation in hydrogel matrix aimed to conduct and maintain minimally required the level of nutrients perfusion for all cells in the expanding organoid. The network configuration should be designed properly in order to exclude hypoxic and necrotic zones in expanding organoid at all stages of its cultivation. In vitro vascularization is currently the main issue within the field of tissue engineering. As perfusion and oxygen transport have direct effects on cell viability and differentiation, researchers are currently limited only to tissues of few millimeters in thickness. These limitations are imposed by mass transfer and are defined by the balance between the metabolic demand of the cellular components in the system and the size of the scaffold. Current approaches include growth factor delivery, channeled scaffolds, perfusion bioreactors, microfluidics, cell co-cultures, cell functionalization, modular assembly, and in vivo systems. These approaches may improve cell viability or generate capillary-like structures within a tissue construct. Thus, there is a fundamental disconnect between defining the metabolic needs of tissue through quantitative measurements of oxygen and nutrient diffusion and the potential ease of integration into host vasculature for future in vivo implantation. A model is proposed for growth prognosis of the organoid perfusion based on joint simulations of general nutrient diffusion, nutrient diffusion to the hydrogel matrix through the contact surfaces and microchannels walls, nutrient consumption by the cells of expanding organoid, including biomatrix contraction during tissue development, which is associated with changed consumption rate of growing organoid cells. The model allows computing effective microchannel network design giving minimally required the level of nutrients concentration in all parts of growing organoid. It can be used for preliminary planning of microchannel network design and simulations of nutrients supply rate depending on the stage of organoid development.

Keywords: 3D model, consumption model, diffusion, spheroid, tissue organoid

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58 Urinary Incontinence and Performance in Elite Athletes

Authors: María Barbaño Acevedo Gómez, Elena Sonsoles Rodríguez López, Sofía Olivia Calvo Moreno, Ángel Basas García, Christophe RamíRez Parenteau

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Introduction: Urinary incontinence (UI) is defined as the involuntary leakage of urine. In persons who practice sport, its prevalence is 36.1% (95% CI 26.5% –46.8%) and varies as it seems to depend on the intensity of exercise, movements and impact on the ground. Such high impact sports are likely to generate higher intra-abdominal pressures and leading to pelvic floor muscle weakness. Although physical exercise reduces the risk of suffering from many diseases the mentality of an elite athlete is not to optimize their health, achieving their goals can put their health at risk. Furthermore, feeling or suffering from any discomfort during training seems to be normal within the elite sport demands. Objective: The main objective of the present study was to know the effects of UI in sports performance in athletes. Methods: This was an observational study conducted in 754 elite athletes. After collecting questions about pelvic floor, UI and sport-related data, participants completed the questionnaire International Consultation on Incontinence Questionnaire-UI Short- Form (ICIQ-SF) and ISI (index of incontinence severity). Results: 48.8% of the athletes declare having losses also in rest, preseason and / or competition (χ2 [3] = 3.64; p = 0.302), being the competition period (29.1%) the most frequent where suffer from urine leakage. Of the elite athletes surveyed, 33% had UI according ICIQ-SF (mean age 23.75 ± 7.74 years). Elite athletes with UI (5.31 ± 1.07 days) dedicate significantly more days per week to training [M = 0.28; 95% CI = 0.08-0.48; t (752) = 2.78; p = 0.005] than those without UI. Regarding frequency, 59.7% lose urine once a week, 25.6% lose urine more than 3 times a week, and 14.7% daily. Based on the amount, approximately 15% claim to lose a moderate and abundant. Athletes with the highest number of urine leaks during their training, the UI affects them more in their daily life (r = 0.259; p = 0.001), they present a greater number of losses in their day to day (r = 0.341; p <0.001 ) and greater severity of UI (r = 0.341; p <0.001). Conclusions: Athletes consider that UI affects them negatively in their daily routine, 30.9% affirm having a severity between moderate and severe in their daily routine, and 29.1% loss urine in competition period. An interesting fact is that more than half of the samples collected were elite athletes who compete at the highest level (Olympic Games, World and European Championship), the dedication to sport occupies a big piece in their life. The most frequent period where athletes suffers urine leakage is in competition and there are many emotions that athletes manage to get their best performance, if we add urine losses in that moments it is possible that their performance could be affected.

Keywords: athletes, performance, prevalence, sport, training, urinary incontinence

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57 Gas-Phase Nondestructive and Environmentally Friendly Covalent Functionalization of Graphene Oxide Paper with Amines

Authors: Natalia Alzate-Carvajal, Diego A. Acevedo-Guzman, Victor Meza-Laguna, Mario H. Farias, Luis A. Perez-Rey, Edgar Abarca-Morales, Victor A. Garcia-Ramirez, Vladimir A. Basiuk, Elena V. Basiuk

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Direct covalent functionalization of prefabricated free-standing graphene oxide paper (GOP) is considered as the only approach suitable for systematic tuning of thermal, mechanical and electronic characteristics of this important class of carbon nanomaterials. At the same time, the traditional liquid-phase functionalization protocols can compromise physical integrity of the paper-like material up to its total disintegration. To avoid such undesirable effects, we explored the possibility of employing an alternative, solvent-free strategy for facile and nondestructive functionalization of GOP with two representative aliphatic amines, 1-octadecylamine (ODA) and 1,12-diaminododecane (DAD), as well as with two aromatic amines, 1-aminopyrene (AP) and 1,5-diaminonaphthalene (DAN). The functionalization was performed under moderate heating at 150-180 °C in vacuum. Under such conditions, it proceeds through both amidation and epoxy ring opening reactions. Comparative characterization of pristine and amine-functionalized GOP mats was carried out by using Fourier-transform infrared, Raman, and X-ray photoelectron spectroscopy (XPS), thermogravimetric (TGA) and differential thermal analysis, scanning electron and atomic force microscopy (SEM and AFM, respectively). Besides that, we compared the stability in water, wettability, electrical conductivity and elastic (Young's) modulus of GOP mats before and after amine functionalization. The highest content of organic species was obtained in the case of GOP-ODA, followed by GOP-DAD, GOP-AP and GOP-DAN samples. The covalent functionalization increased mechanical and thermal stability of GOP, as well as its electrical conductivity. The magnitude of each effect depends on the particular chemical structure of amine employed, which allows for tuning a given GOP property. Morphological characterization by using SEM showed that, compared to pristine graphene oxide paper, amine-modified GOP mats become relatively ordered layered assemblies, in which individual GO sheets are organized in a near-parallel pattern. Financial support from the National Autonomous University of Mexico (grants DGAPA-IN101118 and IN200516) and from the National Council of Science and Technology of Mexico (CONACYT, grant 250655) is greatly appreciated. The authors also thank David A. Domínguez (CNyN of UNAM) for XPS measurements and Dr. Edgar Alvarez-Zauco (Faculty of Science of UNAM) for the opportunity to use TGA equipment.

Keywords: amines, covalent functionalization, gas-phase, graphene oxide paper

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56 Noninvasive Technique for Measurement of Heartbeat in Zebrafish Embryos Exposed to Electromagnetic Fields at 27 GHz

Authors: Sara Ignoto, Elena M. Scalisi, Carmen Sica, Martina Contino, Greta Ferruggia, Antonio Salvaggio, Santi C. Pavone, Gino Sorbello, Loreto Di Donato, Roberta Pecoraro, Maria V. Brundo

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The new fifth generation technology (5G), which should favor high data-rate connections (1Gbps) and latency times lower than the current ones (<1ms), has the characteristic of working on different frequency bands of the radio wave spectrum (700 MHz, 3.6-3.8 GHz and 26.5-27.5 GHz), thus also exploiting higher frequencies than previous mobile radio generations (1G-4G). The higher frequency waves, however, have a lower capacity to propagate in free space and therefore, in order to guarantee the capillary coverage of the territory for high reliability applications, it will be necessary to install a large number of repeaters. Following the introduction of this new technology, there has been growing concern in recent years about the possible harmful effects on human health and several studies were published using several animal models. This study aimed to observe the possible short-term effects induced by 5G-millimeter waves on heartbeat of early life stages of Danio rerio using DanioScope software (Noldus). DanioScope is the complete toolbox for measurements on zebrafish embryos and larvae. The effect of substances can be measured on the developing zebrafish embryo by a range of parameters: earliest activity of the embryo’s tail, activity of the developing heart, speed of blood flowing through the vein, length and diameters of body parts. Activity measurements, cardiovascular data, blood flow data and morphometric parameters can be combined in one single tool. Obtained data are elaborate and provided by the software both numerical as well as graphical. The experiments were performed at 27 GHz by a no commercial high gain pyramidal horn antenna. According to OECD guidelines, exposure to 5G-millimeter waves was tested by fish embryo toxicity test within 96 hours post fertilization, Observations were recorded every 24h, until the end of the short-term test (96h). The results have showed an increase of heartbeat rate on exposed embryos at 48h hpf than control group, but this increase has not been shown at 72-96 h hpf. Nowadays, there is a scant of literature data about this topic, so these results could be useful to approach new studies and also to evaluate potential cardiotoxic effects of mobile radiofrequency.

Keywords: Danio rerio, DanioScope, cardiotoxicity, millimeter waves.

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55 The 2017 Summer Campaign for Night Sky Brightness Measurements on the Tuscan Coast

Authors: Andrea Giacomelli, Luciano Massetti, Elena Maggi, Antonio Raschi

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The presentation will report the activities managed during the Summer of 2017 by a team composed by staff from a University Department, a National Research Council Institute, and an outreach NGO, collecting measurements of night sky brightness and other information on artificial lighting, in order to characterize light pollution issues on portions of the Tuscan coast, in Central Italy. These activities combine measurements collected by the principal scientists, citizen science observations led by students, and outreach events targeting a broad audience. This campaign aggregates the efforts of three actors: the BuioMetria Partecipativa project, which started collecting light pollution data on a national scale in 2008 with an environmental engineering and free/open source GIS core team; the Institute of Biometeorology from the National Research Council, with ongoing studies on light and urban vegetation and a consolidated track record in environmental education and citizen science; the Department of Biology from the University of Pisa, which started experiments to assess the impact of light pollution in coastal environments in 2015. While the core of the activities concerns in situ data, the campaign will account also for remote sensing data, thus considering heterogeneous data sources. The aim of the campaign is twofold: (1) To test actions of citizen and student engagement in monitoring sky brightness (2) To collect night sky brightness data and test a protocol for applications to studies on the ecological impact of light pollution, with a special focus on marine coastal ecosystems. The collaboration of an interdisciplinary team in the study of artificial lighting issues is not a common case in Italy, and the possibility of undertaking the campaign in Tuscany has the added value of operating in one of the territories where it is possible to observe both sites with extremely high lighting levels, and areas with extremely low light pollution, especially in the Southern part of the region. Combining environmental monitoring and communication actions in the context of the campaign, this effort will contribute to the promotion of night skies with a good quality as an important asset for the sustainability of coastal ecosystems, as well as to increase citizen awareness through star gazing, night photography and actively participating in field campaign measurements.

Keywords: citizen science, light pollution, marine coastal biodiversity, environmental education

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54 Effect of Hypoxia on AOX2 Expression in Chlamydomonas reinhardtii

Authors: Maria Ostroukhova, Zhanneta Zalutskaya, Elena Ermilova

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The alternative oxidase (AOX) mediates cyanide-resistant respiration, which bypasses proton-pumping complexes III and IV of the cytochrome pathway to directly transfer electrons from reduced ubiquinone to molecular oxygen. In Chlamydomonas reinhardtii, AOX is a monomeric protein that is encoded by two genes of discrete subfamilies, AOX1 and AOX2. Although AOX has been proposed to play essential roles in stress tolerance of organisms, the role of subfamily AOX2 is largely unknown. In C. reinhardtii, AOX2 was initially identified as one of constitutively low expressed genes. Like other photosynthetic organisms C. reinhardtii cells frequently experience periods of hypoxia. To examine AOX2 transcriptional regulation and role of AOX2 in hypoxia adaptation, real-time PCR analysis and artificial microRNA method were employed. Two experimental approaches have been used to induce the anoxic conditions: dark-anaerobic and light-anaerobic conditions. C. reinhardtii cells exposed to the oxygen deprivation have shown increased AOX2 mRNA levels. By contrast, AOX1 was not an anoxia-responsive gene. In C. reinhardtii, a subset of genes is regulated by transcription factor CRR1 in anaerobic conditions. Notable, the AOX2 promoter region contains the potential motif for CRR1 binding. Therefore, the role of CRR1 in the control of AOX2 transcription was tested. The CRR1-underexpressing strains, that were generated and characterized in this work, exhibited low levels of AOX2 transcripts under anoxic conditions. However, the transformants still slightly induced AOX2 gene expression in the darkness. These confirmed our suggestions that darkness is a regulatory stimulus for AOX genes in C. reinhardtii. Thus, other factors must contribute to AOX2 promoter activity under dark-anoxic conditions. Moreover, knock-down of CRR1 caused a complete reduction of AOX2 expression under light-anoxic conditions. These results indicate that (1) CRR1 is required for AOX2 expression during hypoxia, and (2) AOX2 gene is regulated by CRR1 together with yet-unknown regulatory factor(s). In addition, the AOX2-underexpressing strains were generated. The analysis of amiRNA-AOX2 strains suggested a role of this alternative oxidase in hypoxia adaptation of the alga. In conclusion, the results reported here show that C. reinhardtii AOX2 gene is stress inducible. CRR1 transcriptional factor is involved in the regulation of the AOX2 gene expression in the absence of oxygen. Moreover, AOX2 but not AOX1 functions under oxygen deprivation. This work was supported by Russian Science Foundation (research grant № 16-14-10004).

Keywords: alternative oxidase 2, artificial microRNA approach, chlamydomonas reinhardtii, hypoxia

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53 In Vivo Evaluation of Exposure to Electromagnetic Fields at 27 GHz (5G) of Danio Rerio: A Preliminary Study

Authors: Elena Maria Scalisi, Roberta Pecoraro, Martina Contino, Sara Ignoto, Carmelo Iaria, Santi Concetto Pavone, Gino Sorbello, Loreto Di Donato, Maria Violetta Brundo

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5G Technology is evolving to satisfy a variety of service requirements that may allow high data-rate connections (1Gbps) and lower latency times than current (<1ms). In order to support a high data transmission speed and a high traffic service for eMBB (enhanced mobile broadband) use cases, 5G systems have the characteristic of using different frequency bands of the radio wave spectrum (700 MHz, 3.6-3.8 GHz and 26.5-27.5 GHz), thus taking advantage of higher frequencies than previous mobile radio generations (1G-4G). However, waves at higher frequencies have a lower capacity to propagate in free space and therefore, in order to guarantee the capillary coverage of the territory for high reliability applications, it will be necessary to install a large number of repeaters. Following the introduction of this new technology, there has been growing concern over the past few months about possible harmful effects on human health. The aim of this preliminary study is to evaluate possible short term effects induced by 5G-millimeter waves on embryonic development and early life stages of Danio rerio by Z-FET. We exposed developing zebrafish at frequency of 27 GHz, with a standard pyramidal horn antenna placed at 15 cm far from the samples holder ensuring an incident power density of 10 mW/cm2. During the exposure cycle, from 6 h post fertilization (hpf) to 96 hpf, we measured a different morphological endpoints every 24 hours. Zebrafish embryo toxicity test (Z-FET) is a short term test, carried out on fertilized eggs of zebrafish and it represents an effective alternative to acute test with adult fish (OECD, 2013). We have observed that 5G did not reveal significant impacts on mortality nor on morphology because exposed larvae showed a normal detachment of the tail, presence of heartbeat, well-organized somites, therefore hatching rate was lower than untreated larvae even at 48 h of exposure. Moreover, the immunohistochemical analysis performed on larvae showed a negativity to the HSP-70 expression used as a biomarkers. This is a preliminary study on evaluation of potential toxicity induced by 5G and it seems appropriate to underline the importance that further studies would take, aimed at clarifying the probable real risk of exposure to electromagnetic fields.

Keywords: Biomarker of exposure, embryonic development, 5G waves, zebrafish embryo toxicity test

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52 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

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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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51 The Gypsy Community Facing the Sexual Orientation: An Empirical Approach to the Attitudes of the Gypsy Population of Granada Towards Homosexual Sex-Affective Relationships

Authors: Elena Arquer Cuenca

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The gypsy community has been a mistreated and rejected group since its arrival in the Iberian Peninsula in the 15th century. At present, despite being the largest ethnic minority group in Spain as well as in Europe, the different legal and social initiatives in favour of equality continue to suffer discrimination by the general society. This has fostered a strengthening of the endogroup accompanied by cultural conservatism as a form of self-protection. Despite the current trend of normalization of sexual diversity in modern societies, LGB people continue to suffer discrimination, especially in more traditional environments or communities. This rejection for reasons of sexual orientation within the family or community can hinder the free development of the person and compromise peaceful coexistence. The present work is intended as an approach to the attitudes of the gypsy population towards non-heterosexual sexual orientation. The objective is none other than ‘to know the appreciation that the gypsy population has about homosexual sex-affective relationships, in order to assess whether this has any impact on family and community coexistence’. The following specific objectives are derived from this general objective: ‘To find out whether there is a relationship between the dichotomous Roma gender system and the acceptance/rejection of homosexuality’; ‘to analyse whether sexual orientation has an impact on the coexistence of the Roman family and community’; ‘to analyse whether the historical discrimination suffered by the Roman population favours the maintenance of the patriarchal heterosexual reproductive family’; and lastly ‘to explore whether ICTs have promoted the process of normalisation and/or acceptance of homosexuality within the Roma community’. In order to achieve these objectives, a bibliographical and documentary review has been used, as well as the semi-structured interview technique, in which 4 gypsy people participated (2 women and 2 men of different ages). One of the main findings was the inappropriateness of the use of the homogenising category "Gypsy People" at present, given the great diversity among the Roma communities. Moreover, the difficulty in accepting homosexuality seems to be related to the fact that the heterosexual reproductive family has been the main survival mechanism of Roma communities over centuries. However, it will be concluded that attitudes towards homosexuality will vary depending on the socio-economic and cultural context and factors such as age or professed religion. Three main contributions of this research are: firstly, the inclusion of sexual orientation as a variable to be considered when analysing peaceful coexistence; secondly socio-historical dynamics and structures of inequality have been taken into account when analysing Roma attitudes towards homosexuality; and finally, the processual nature of socio-cultural changes has also been considered.

Keywords: gender, homosexuality, ICTs, peaceful coexistence, Roma community, sexual orientation

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50 Paradigms of Sustainability: Roles and Impact of Communication in the Fashion System

Authors: Elena Pucci, Margherita Tufarelli, Leonardo Giliberti

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As central for human and social development of the future, sustainability is becoming a recurring theme also in the fashion industry, where the need to explore new possible directions aimed at achieving sustainability goals and their communication is rising. Scholars have been devoted to the overall environmental impact of the textile and fashion industry, which, emerging as one of the world’s most polluting, today concretely assumes the need to take the path of sustainability in both products and production processes. Every day we witness the impact of our consumption, showing that the sustainability concept is as vast as complex: with a sometimes ambiguous definition, sustainability can concern projects, products, companies, sales, packagings, supply chains in relation to the actors proximity as well as traceability, raw materials procurement, and disposal. However, in its primary meaning, sustainability is the ability to maintain specific values and resources for future generations. The contribution aims to address sustainability in the fashion system as a layered problem that requires substantial changes at different levels: in the fashion product (materials, production processes, timing, distribution, and disposal), in the functioning of the system (life cycle, impact, needs, communication) and last but not least in the practice of fashion design which should conceive durable, low obsolescence and possibly demountable products. Moreover, consumers play a central role for the growing awareness, together with an increasingly strong sensitivity towards the environment and sustainable clothing. Since it is also a market demand, undertaking significant efforts to achieve total transparency and sustainability in all production and distribution processes is becoming fundamental for the fashion system. Sustainability is not to be understood as purely environmental but as the pursuit of collective well-being in relation to conscious production, human rights, and social dignity with the aim to achieve intelligent, resource, and environmentally friendly production and consumption patterns. Assuming sustainability as a layered problem makes the role of communication crucial to convey scientific or production specific content so that people can obtain and interpret information to make related decisions. Hence, if it is true that “what designers make becomes the future we inhabit'', design is facing great and challenging responsibility. The fashion industry needs a system of rules able to assess the sustainability of products, which is transparent and easily interpreted by consumers, identifying and enhancing virtuous practices. There are still complex and fragmented value chains that make it extremely difficult for brands and manufacturers to know the history of their products, to identify exactly where the risks lie, and to respond to the growing demand from consumers and civil society for responsible and sustainable production practices in the fashion industry.

Keywords: fashion design, fashion system, sustainability, communication, complexity

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49 Impact of Helicobacter pylori Infection on Colorectal Adenoma-Colorectal Carcinoma Sequence

Authors: Jannis Kountouras, Nikolaos Kapetanakis, Stergios A. Polyzos, Apostolis Papaeftymiou, Panagiotis Katsinelos, Ioannis Venizelos, Christina Nikolaidou, Christos Zavos, Iordanis Romiopoulos, Elena Tsiaousi, Evangelos Kazakos, Michael Doulberis

Abstract:

Background & Aims: Helicobacter pylori infection (Hp-I) has been recognized as a substantial risk agent involved in gastrointestinal (GI) tract oncogenesis by stimulating cancer stem cells (CSCs), oncogenes, immune surveillance processes, and triggering GI microbiota dysbiosis. We aimed to investigate the possible involvement of active Hp-I in the sequence: chronic inflammation–adenoma–colorectal cancer (CRC) development. Methods: Four pillars were investigated: (i) endoscopic and conventional histological examinations of patients with CRC, colorectal adenomas (CRA) versus controls to detect the presence of active Hp-I; (ii) immunohistochemical determination of the presence of Hp; expression of CD44, an indicator of CSCs and/or bone marrow-derived stem cells (BMDSCs); expressions of oncogene Ki67 and anti-apoptotic Bcl-2 protein; (iii) expression of CD45, indicator of immune surveillance locally (assessing mainly T and B lymphocytes locally); and (iv) correlation of the studied parameters with the presence or absence of Hp-I. Results: Among 50 patients with CRC, 25 with CRA, and 10 controls, a significantly higher presence of Hp-I in the CRA (68%) and CRC group (84%) were found compared with controls (30%). The presence of Hp-I with accompanying immunohistochemical expression of CD44 in biopsy specimens was revealed in a high proportion of patients with CRA associated with moderate/severe dysplasia (88%) and CRC patients with moderate/severe degree of malignancy (91%). Comparable results were also obtained for Ki67, Bcl-2, and CD45 immunohistochemical expressions. Concluding Remarks: Hp-I seems to be involved in the sequence: CRA – dysplasia – CRC, similarly to the upper GI tract oncogenesis, by several pathways such as the following: Beyond Hp-I associated insulin resistance, the major underlying mechanism responsible for the metabolic syndrome (MetS) that increase the risk of colorectal neoplasms, as implied by other Hp-I related MetS pathologies, such as non-alcoholic fatty liver disease and upper GI cancer, the disturbance of the normal GI microbiota (i.e., dysbiosis) and the formation of an irritative biofilm could contribute to a perpetual inflammatory upper GIT and colon mucosal damage, stimulating CSCs or recruiting BMDSCs and affecting oncogenes and immune surveillance processes. Further large-scale relative studies with a pathophysiological perspective are necessary to demonstrate in-depth this relationship.

Keywords: Helicobacter pylori, colorectal cancer, colorectal adenomas, gastrointestinal oncogenesis

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48 Recognition by the Voice and Speech Features of the Emotional State of Children by Adults and Automatically

Authors: Elena E. Lyakso, Olga V. Frolova, Yuri N. Matveev, Aleksey S. Grigorev, Alexander S. Nikolaev, Viktor A. Gorodnyi

Abstract:

The study of the children’s emotional sphere depending on age and psychoneurological state is of great importance for the design of educational programs for children and their social adaptation. Atypical development may be accompanied by violations or specificities of the emotional sphere. To study characteristics of the emotional state reflection in the voice and speech features of children, the perceptual study with the participation of adults and the automatic recognition of speech were conducted. Speech of children with typical development (TD), with Down syndrome (DS), and with autism spectrum disorders (ASD) aged 6-12 years was recorded. To obtain emotional speech in children, model situations were created, including a dialogue between the child and the experimenter containing questions that can cause various emotional states in the child and playing with a standard set of toys. The questions and toys were selected, taking into account the child’s age, developmental characteristics, and speech skills. For the perceptual experiment by adults, test sequences containing speech material of 30 children: TD, DS, and ASD were created. The listeners were 100 adults (age 19.3 ± 2.3 years). The listeners were tasked with determining the children’s emotional state as “comfort – neutral – discomfort” while listening to the test material. Spectrographic analysis of speech signals was conducted. For automatic recognition of the emotional state, 6594 speech files containing speech material of children were prepared. Automatic recognition of three states, “comfort – neutral – discomfort,” was performed using automatically extracted from the set of acoustic features - the Geneva Minimalistic Acoustic Parameter Set (GeMAPS) and the extended Geneva Minimalistic Acoustic Parameter Set (eGeMAPS). The results showed that the emotional state is worse determined by the speech of TD children (comfort – 58% of correct answers, discomfort – 56%). Listeners better recognized discomfort in children with ASD and DS (78% of answers) than comfort (70% and 67%, respectively, for children with DS and ASD). The neutral state is better recognized by the speech of children with ASD (67%) than by the speech of children with DS (52%) and TD children (54%). According to the automatic recognition data using the acoustic feature set GeMAPSv01b, the accuracy of automatic recognition of emotional states for children with ASD is 0.687; children with DS – 0.725; TD children – 0.641. When using the acoustic feature set eGeMAPSv01b, the accuracy of automatic recognition of emotional states for children with ASD is 0.671; children with DS – 0.717; TD children – 0.631. The use of different models showed similar results, with better recognition of emotional states by the speech of children with DS than by the speech of children with ASD. The state of comfort is automatically determined better by the speech of TD children (precision – 0.546) and children with ASD (0.523), discomfort – children with DS (0.504). The data on the specificities of recognition by adults of the children’s emotional state by their speech may be used in recruitment for working with children with atypical development. Automatic recognition data can be used to create alternative communication systems and automatic human-computer interfaces for social-emotional learning. Acknowledgment: This work was financially supported by the Russian Science Foundation (project 18-18-00063).

Keywords: autism spectrum disorders, automatic recognition of speech, child’s emotional speech, Down syndrome, perceptual experiment

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47 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

Abstract:

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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46 Survival Analysis after a First Ischaemic Stroke Event: A Case-Control Study in the Adult Population of England.

Authors: Padma Chutoo, Elena Kulinskaya, Ilyas Bakbergenuly, Nicholas Steel, Dmitri Pchejetski

Abstract:

Stroke is associated with a significant risk of morbidity and mortality. There is scarcity of research on the long-term survival after first-ever ischaemic stroke (IS) events in England with regards to effects of different medical therapies and comorbidities. The objective of this study was to model the all-cause mortality after an IS diagnosis in the adult population of England. Using a retrospective case-control design, we extracted the electronic medical records of patients born prior to or in year 1960 in England with a first-ever ischaemic stroke diagnosis from January 1986 to January 2017 within the Health and Improvement Network (THIN) database. Participants with a history of ischaemic stroke were matched to 3 controls by sex and age at diagnosis and general practice. The primary outcome was the all-cause mortality. The hazards of the all-cause mortality were estimated using a Weibull-Cox survival model which included both scale and shape effects and a shared random effect of general practice. The model included sex, birth cohort, socio-economic status, comorbidities and medical therapies. 20,250 patients with a history of IS (cases) and 55,519 controls were followed up to 30 years. From 2008 to 2015, the one-year all-cause mortality for the IS patients declined with an absolute change of -0.5%. Preventive treatments to cases increased considerably over time. These included prescriptions of statins and antihypertensives. However, prescriptions for antiplatelet drugs decreased in the routine general practice since 2010. The survival model revealed a survival benefit of antiplatelet treatment to stroke survivors with hazard ratio (HR) of 0.92 (0.90 – 0.94). IS diagnosis had significant interactions with gender and age at entry and hypertension diagnosis. IS diagnosis was associated with high risk of all-cause mortality with HR= 3.39 (3.05-3.72) for cases compared to controls. Hypertension was associated with poor survival with HR = 4.79 (4.49 - 5.09) for hypertensive cases relative to non-hypertensive controls, though the detrimental effect of hypertension has not reached significance for hypertensive controls, HR = 1.19(0.82-1.56). This study of English primary care data showed that between 2008 and 2015, the rates of prescriptions of stroke preventive treatments increased, and a short-term all-cause mortality after IS stroke declined. However, stroke resulted in poor long-term survival. Hypertension, a modifiable risk factor, was found to be associated with poor survival outcomes in IS patients. Antiplatelet drugs were found to be protective to survival. Better efforts are required to reduce the burden of stroke through health service development and primary prevention.

Keywords: general practice, hazard ratio, health improvement network (THIN), ischaemic stroke, multiple imputation, Weibull-Cox model.

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