Search results for: Munoz A. Tatiana

Authors: Munoz A. Tatiana, Solano R. Brandon, Montes C. Juan, Cierco G. Javier

Abstract:

The hydrodynamic cavitation is a process in which the energy that the fluids have in the phase changes is used. From this energy, local temperatures greater than 5000 °C are obtained where thermal cracking of the fluid molecules takes place. The process applied to heavy oil affects variables such as viscosity, density, and composition, which constitutes an important improvement in the quality of crude oil. In this study, the need to design a software through mathematical integration models of mixing, cavitation, kinetics, and reactor, allows modeling changes in density, viscosity, and composition of a heavy oil crude, when the fluid passes through a hydrodynamic cavitation reactor. In order to evaluate the viability of this technique in the industry, a heavy oil of 18° API gravity, was simulated using naphtha as a hydrogen donor at concentrations of 1, 2 and 5% vol, where the simulation results showed an API gravity increase to 0.77, 1.21 and 1.93° respectively and a reduction viscosity by 9.9, 12.9 and 15.8%. The obtained results allow to have a favorable panorama on this technological development, an appropriate visualization on the generation of innovative knowledge of this technique and the technical-economic opportunity that benefits the development of the hydrocarbon sector related to heavy crude oil that includes the largest world oil production.

Keywords: hydrodynamic cavitation, thermal cracking, hydrogen donor, heavy oil upgrading, simulator

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

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EEG correlates of mathematical and trait anxiety level were studied in 52 healthy Russian-speakers during execution of error-recognition tasks with lexical, arithmetic and algebraic conditions. Event-related spectral perturbations were used as a measure of brain activity. The ERSP plots revealed 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 conditions. The correlates of anxiety were found in theta (4-8 Hz) and beta2 (16-20 Hz) frequency bands. In theta band the effects of mathematical anxiety were stronger expressed in lexical, than in arithmetic and algebraic condition. The mathematical anxiety effects in theta band were associated with differences between anterior and posterior cortical areas, whereas the effects of trait anxiety were associated with inter-hemispherical differences. In beta1 and beta2 bands effects of trait and mathematical anxiety were directed oppositely. The trait anxiety was associated with increase of amplitude of desynchronization, whereas the mathematical anxiety was associated with decrease of this amplitude. The effect of mathematical anxiety in beta2 band was insignificant for lexical condition but was the strongest in algebraic condition. EEG correlates of anxiety in theta band could be interpreted as indexes of task emotionality, whereas the reaction in beta2 band is related to tension of intellectual resources.

Keywords: EEG, brain activity, lexical and numerical error-recognition tasks, mathematical and trait anxiety

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Authors: Samaniego Campoverde José Enrique, Rosero Muñoz Jorge Enrique, Luzcando Narea Lorena Elizabeth

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This paper presents a practical and theoretical model for the operation of the Tesla coil using digital signal processing. The research is based on the analysis of ten scientific papers exploring the development and operation of the Tesla coil. Starting from the Testa coil, several modifications were carried out on the Tesla coil, with the aim of amplifying the digital signal by making use of digital signal processing. To achieve this, an amplifier with a transistor and digital filters provided by MATLAB software were used, which were chosen according to the characteristics of the signals in question.

Keywords: tesla coil, digital signal process, equalizer, graphical environment

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Authors: Daria N. Podvigina, Tatiana V. Chernigovskaya

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We have studied the temporal characteristics of bistable perception of the stimuli of two types: one involves alterations in a perceived depth and another one has an ambiguous content. We used the Necker lattice and lines of shadowed circles ambiguously perceived either as spheres or holes as stimuli of the first type. The Winson figure (the Eskimo/Indian picture) was a stimulus of the second type. We have analyzed how often the reversals occurred (reversal rate) and for how long each of the two interpretations, or percepts, was observed during one presentation (stability durations). For all three ambiguous images the reversal rate and the stability durations had similar values, which provide another evidence for a significant role of top-down processes in multistable perception.

Keywords: multistable perception, perceived depth, reversal rate, top-down processes

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Authors: Mario Lupaca, Karol Munoz, Victor De Negri

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The present article presents a methodology for the improvement of the energy efficiency in pneumatic systems through the restoring of air. In this way, three techniques of expansion of a cylinder are identified: Expansion using the air of the compressor (conventional), restoring the air (efficient), and combining the air of the compressor and the restored air (hybrid). The methodology starts with the development of the GRAFCET of the system so that it can be decided whether to expand the cylinder in a conventional, efficient, or hybrid way. The methodology can be applied to any case. Finally, graphs of comparison between the three methods of expansion with certain cylinder strokes and workloads are presented, to facilitate the subsequent selection of one system or another.

Keywords: energetic, efficiency, GRAFCET, methodology, pneumatic

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Authors: Sergey A. Burikov, Tatiana A. Dolenko, Kirill A. Gushchin, Sergey A. Dolenko

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The paper presents the results of clusterization by Kohonen self-organizing maps (SOM) applied for analysis of array of Raman spectra of multi-component solutions of inorganic salts, for determination of types of salts present in the solution. It is demonstrated that use of SOM is a promising method for solution of clusterization and classification problems in spectroscopy of multi-component objects, as attributing a pattern to some cluster may be used for recognition of component composition of the object.

Keywords: Kohonen self-organizing maps, clusterization, multi-component solutions, Raman spectroscopy

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Authors: F. Boudet, V. Galano, D. Gmira, L. Munoz, A. Reina

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Commodities pricing needs a specific approach as they are often linked to each other and so are expectedly doing their prices. They are called co-integrated when at least one stationary linear combination exists between them. Though widespread in economic literature, and even if many equilibrium relations and co-movements exist in the economy, this principle of co-movement is not developed in derivatives field. The present study focuses on the following problem: How can the price of a forward agreement on a commodity be simulated, when it is co-integrated with other ones? Theoretical analysis is developed from Gibson-Schwartz model and an analytical solution is given for short maturities contracts and under risk-neutral conditions. The application has been made to crude oil and heating oil energy commodities and result confirms the applicability of proposed method.

Keywords: co-integration, commodities, forward pricing, Gibson-Schwartz

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Authors: Encarnación Álvarez, Rosa M. García-Fernández, Francisco J. Blanco-Encomienda, Juan F. Muñoz

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The European Union Survey on Income and Living Conditions (EU-SILC) is a popular survey which provides information on income, poverty, social exclusion and living conditions of households and individuals in the European Union. The EUSILC contains variables which may contain outliers. The presence of outliers can have an impact on the measures and indicators used by the EU-SILC. In this paper, we used data sets from various countries to analyze the presence of outliers. In addition, we obtain some indicators after removing these outliers, and a comparison between both situations can be observed. Finally, some conclusions are obtained.

Keywords: poverty line, headcount index, risk of poverty, skewness coefficient

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Authors: Víctor Manuel Muñoz-Sánchez, Antonio Manuel Pérez-Flores

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This paper analyzes the institutionalization of social protest in Spain. In the current crisis Podemos party seems to represent the political positions of the most affected citizens by the economic situation. It studies using quantitative techniques (statistical bivariate analysis), focusing on the exploitation of several bases of statistics data from the Center for Sociological and Research of Spanish Government, 15M movement characterization to its institutionalization in the Podemos party. Making a comparison between the participant's profile by the 15M and the social bases of Podemos votes. Data on the transformation of the socio-demographic profile of the fans, connoisseurs and 15M participants and voters are given.

Keywords: collective action, emerging parties, political parties, social protest

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Authors: Encarnación Álvarez, Rosa M. García-Fernández, Juan F. Muñoz, Francisco J. Blanco-Encomienda

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The problem of estimating a proportion has important applications in the field of economics, and in general, in many areas such as social sciences. A common application in economics is the estimation of the headcount index. In this paper, we define the general headcount index as a proportion. Furthermore, we introduce a new quantitative method for estimating the headcount index. In particular, we suggest to use the logistic regression estimator for the problem of estimating the headcount index. Assuming a real data set, results derived from Monte Carlo simulation studies indicate that the logistic regression estimator can be more accurate than the traditional estimator of the headcount index.

Keywords: poverty line, poor, risk of poverty, Monte Carlo simulations, sample

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Authors: Tatiana V. Korsakova

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The cognitive methodology in management is considered: Cognitive structuring - the formation of ideas about the functioning of a developing organization; Cognitive modeling - heuristic construction of existing actions (zone of successful actions); and Cognitive construct - the formation of filters for converting external information into specific events of managerial reality. The major findings of the study are the identification of areas of successful actions in the organization, harmonization of criteria for evaluating the effectiveness of company management, and the frame-description that indicates the connection of environmental elements with the elements of the organization. It is stated the development of specific events of managerial reality in the direction of the further development of the organization depends on the personal cognitive construct of the development-subjects when it is used in the zone of successful actions.

Keywords: cognitive construct, focus of applicability, knowledge corporate culture, zones of successful actions

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Authors: Indra Gonzalez Ojeda, Tatiana Quinones, Manuel Rosario, Igor Lednev, Juan Lopez Garriga

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Amyloid fibrils are stable aggregates of misfolded protein associated with many neurodegenerative disorders. It has been shown that hydrogen sulfide (H2S), inhibits the fibrillation of lysozyme through the formation of trisulfide (S-S-S) bonds. However, the overall mechanism remains elusive. Here, the concentration dependence of H2S effect was investigated using Atomic force microscopy (AFM), non-resonance Raman spectroscopy, Deep-UV Raman spectroscopy and circular dichroism (CD). It was found that small spherical aggregates with trisulfide bonds and a unique secondary structure were formed instead of amyloid fibrils when adding concentrations of 25 mM and 50 mM of H2S. This could indicate that H2S might serve as a protecting agent for the protein. However, further characterization of these aggregates and their trisulfide bonds is needed to fully unravel the function H2S has on protein fibrillation.

Keywords: amyloid fibrils, hydrogen sulfide, protein folding, raman spectroscopy

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Authors: Jhon Vargas, Gilberto Osorio-Gomez, Tatiana Manrique

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This work presents an optimal driving strategy proposal for a 125 c.c. street-type hybrid electric motorcycle with a parallel configuration. The results presented in this article are complementary regarding the control proposal of a hybrid motorcycle. In order to carry out such developments, a representative dynamic model of the motorcycle is used, in which also are described different optimization functionalities for predetermined driving modes. The purpose is to implement an off-line optimal driving strategy which distributes energy to both engines by minimizing an objective torque requirement function. An optimal dynamic contribution is found from the optimization routine, and the optimal percentage contribution for vehicle cruise speed is implemented in the proposed online PID controller.

Keywords: dynamic model, driving strategies, parallel hybrid motorcycle, PID controller, optimization

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Authors: Tatiana A. Dolenko, Sergey A. Burikov, Alexander O. Efitorov, Sergey A. Dolenko

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In this study, a comparative analysis of the approaches associated with the use of neural network algorithms for effective solution of a complex inverse problem – the problem of identifying and determining the individual concentrations of inorganic salts in multicomponent aqueous solutions by the spectra of Raman scattering of light – is performed. It is shown that application of artificial neural networks provides the average accuracy of determination of concentration of each salt no worse than 0.025 M. The results of comparative analysis of input data compression methods are presented. It is demonstrated that use of uniform aggregation of input features allows decreasing the error of determination of individual concentrations of components by 16-18% on the average.

Keywords: inverse problems, multi-component solutions, neural networks, Raman spectroscopy

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Authors: L. M. López López, J. V. Montesdeoca Contreras, A. R. Cuji Fajardo, L. E. Garzón Muñoz, J. I. Fajardo Seminario

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In this article a new method of cleaning process of metal sheets for household appliances was developed, using low-pressure cold plasma. In this context, this research consist in analyze the results of metal sheets cleaning process using plasma and compare with pickling process to determinate the efficiency of each process and the level of contamination produced. Surface Cleaning was evaluated by measuring the contact angle with deionized water, diiodo methane and ethylene glycol, for the calculus of the surface free energy by means of the Fowkes theories and Wu. Showing that low-pressure cold plasma is very efficient both in cleaning process how in environment impact.

Keywords: efficient use of plasma, ecological impact of plasma, metal sheets cleaning means, plasma cleaning process.

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Authors: Miguel A. Borrella, Ana P. Fanjul, Suca Munoz, Liliana Herrera

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This paper evaluates the impact of LEADER, a remarkable Community-Led Local Development (CLLD) approach of the European Program for Rural Development applied to rural municipalities of Spain in 2018 and 2019. Using a difference-in-difference estimation strategy and a newly-constructed database, results show that aided municipalities have significantly lower unemployment levels than non-aided municipalities. Results are significant for the decrease in unemployment for both women and people younger than 25 years old, two of the target groups of the policy. Nevertheless, they are larger for male and older workers. Therefore, findings suggest that LEADER 2017-2018 was successful in reducing unemployment in rural areas.

Keywords: community-led local development, ex-post evaluation, LEADER, rural development

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Authors: M. Kiarna, S. Junjira, S. Casey, M. Nolwazi, M. S. Marcos, A. T. Tatiana, L. Cassandra

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This paper details a rebrand design project developed for a non-profitable organization called Te Roopu Waiora (TRW), which is currently located in Auckland, Aotearoa New Zealand. This social enterprise is dedicated to supporting the Māori community living with sensorial, physical and intellectual disabilities (whānau hauā). As part of a year three bachelor design brief, the rebrand project enabled students to reflect on Kaupapa Māori principles and appropriately address the values of the organisation. As such, the methodology used a pragmatic paradigm approach and mixed methods design practices involving a human-centred design to problem solving. As result, the student project culminated in the development in a range of cohesive design artefacts, aiming to improve the rentability and perception of the brand with the audience and stakeholders.

Keywords: design in Aotearoa New Zealand, Kaupapa Māori, branding, design education, human-centered design

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Authors: Encarnacion Alvarez, Noemı Hidalgo-Rebollo, Juan F. Munoz, Francisco J. Blanco-Encomienda

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Many companies use the technique named as acceptance sampling which consists on the inspection and decision making regarding products. According to the results derived from this method, the company takes the decision of acceptance or rejection of a product. The acceptance sampling can be applied to the technology management, since the acceptance sampling can be seen as a tool to improve the design planning, operation and control of technological products. The theoretical operating characteristic (OC) curves are widely used when dealing with acceptance sampling. In this paper, we carry out Monte Carlo simulation studies to compare numerically the empirical OC curves derived from the empirical results to the customary theoretical OC curves. We analyze various possible scenarios in such a way that the differences between the empirical and theoretical curves can be observed under different situations.

Keywords: single-sampling plan, lot, Monte Carlo simulation, quality control

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Authors: Muñoz-Sanchez, Victor Manuel, Perez-Flores, Antonio Manuel

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The main objective of this paper is to establish the relationship between models of state organization and the various types of collective identity expressed by the Spanish. The question of nationalism and identity ascription in Spain has always been a topic of special importance due to the presence in that country of territories where the population emits very different opinions of nationalist sentiment than the rest of Spain. The current situation of sovereignty challenge of Catalonia to the central government exemplifies the importance of the subject matter. In order to analyze this process of interrelation, we use a secondary data mining by applying the multiple correspondence analysis technique (MCA). As a main result a typology of four types of expression of collective identity based on models of State organization are shown, which are connected with the party position on this issue.

Keywords: models of organization of the state, nationalism, collective identity, Spain, political parties

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Authors: Raúl González Muñoz, Essam Shehab, Martin Weinitzke, Chris Fowler, Paul Baguley

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Software applications have become crucial for the aerospace industry, providing a wide range of functionalities and capabilities. However, due to the considerable time difference between aircraft and software life cycles, obsolescence has turned into a major challenge for industry in last decades. This paper aims to provide a view on the different causes of software obsolescence within aerospace industry, as well as a perception on the importance of each of them. The key research question addressed is what drives software obsolescence in the aerospace industry, managing large software application portfolios. This question has been addressed by conducting firstly an in depth review of current literature and secondly by arranging an industry workshop with professionals from aerospace and consulting companies. The result is a set of drivers of software obsolescence, distributed among three different environments and several domains. By incorporating monitoring methodologies to assess those software obsolescence drivers, benefits in maintenance efforts and operations disruption avoidance are expected.

Keywords: aerospace industry, obsolescence drivers, software lifecycle, software obsolescence

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Authors: Tatiana Tunon, Gottfried Pestal

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Government agencies responsible for the management of fisheries resources publish many types of grey literature, and these materials are increasingly accessible to the public on agency websites. However, scope and quality vary considerably, and end-users need meta-data about the report series when deciding whether to use the information (e.g. apply the methods, include the results in a systematic review), or when prioritizing materials for archiving (e.g. library holdings, reference databases). A proposed taxonomy for these report series was developed based on a review of 41 report series from 6 government agencies in 4 countries (Canada, New Zealand, Scotland, and United States). Each report series was categorized according to multiple criteria describing peer-review process, content, and purpose. A robust classification tree was then fitted to these descriptions, and the resulting taxonomic groups were used to compare agency output from 4 countries using reports available in their online repositories.

Keywords: classification tree, fisheries, government, grey literature

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Authors: Frank Alexis, Jenny Antonia Rodriguez Leon, Cristobal Leonardo Dominguez Borbor, Mery Rosario Ramirez Munoz

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The shrimp industry has increased in the last years to the point of becoming one of the most dynamic industries. However, the appearance of diseases that significantly affect the production of shrimps has been an obstacle for the shrimp industry. We hypothesized that natural fibers from biodiversity can stimulate the immune system to prevent shrimp diseases like vibriosis. In this project, we extracted the fibers from vegetal sources in Ecuador and characterized them using common techniques like XRD, SEM, and then we tested the effect of fibers as immunostimulants for shrimps in-vitro and in-vivo using small aquarium and large pools. Our results demonstrate that vegetal fibers can significantly increase the survival of shrimps. Moreover, the production of shrimps in a large pool was significantly increased. Lastly, the test of color and taste successfully surpass the control group of shrimps not treated with fiber food supplements.

Keywords: fibers, immunostimulant, shrimp, vibriosis

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Authors: Claudia Marcela Munoz Gonzalez, Diego Fernando Hernandez Losada, Hugo Alberto Herrera Fonseca

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The project owner's role in the public-private investment consists of controlling and verifying the correct execution of the project's objectives and resources. Likewise, it is a discipline little explored in the academic field, whereby this work wishes to contribute with a model of good practices based on the project management methodology proposed by the Project Management Body of Knowledge® Guide. In the same way, highlight what are the controls that an integral project owner should take into account in its exercise and application, through the stages in which its contract runs. This proposal aims to structure its practice and integrate its functions according to a project management methodology. In addition, these practices will be applied in a case study of projects in the agricultural sector, particularly in the construction of irrigation district in Cundinamarca, Colombia.

Keywords: controls, construction of irrigation district, PMBOK®, project owner

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Authors: Encarnación Álvarez-Verdejo, Raúl Amor-Pulido, Pablo J. Moya-Fernández, Juan F. Muñoz-Rosas, Francisco J. Blanco-Encomienda

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Many companies use the statistical tool named as statistical quality control, and which can have a high cost for the companies interested on these statistical tools. The evaluation of the quality of products and services is an important topic, but the reduction of the cost of the implantation of the statistical quality control also has important benefits for the companies. For this reason, it is important to implement a economic design for the various steps included into the statistical quality control. In this paper, we describe some relevant aspects related to the economic design of a quality control chart for the proportion of defective items. They are very important because the suggested issues can reduce the cost of implementing a quality control chart for the proportion of defective items. Note that the main purpose of this chart is to evaluate and control the proportion of defective items of a production process.

Keywords: proportion, type I error, economic plan, distribution function

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Authors: Encarnación Álvarez, Rosa M. García-Fernández, Juan F. Muñoz

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Estimation of a proportion has many applications in economics and social studies. A common application is the estimation of the low income proportion, which gives the proportion of people classified as poor into a population. In this paper, we present this poverty indicator and propose to use the logistic regression estimator for the problem of estimating the low income proportion. Various sampling designs are presented. Assuming a real data set obtained from the European Survey on Income and Living Conditions, Monte Carlo simulation studies are carried out to analyze the empirical performance of the logistic regression estimator under the various sampling designs considered in this paper. Results derived from Monte Carlo simulation studies indicate that the logistic regression estimator can be more accurate than the customary estimator under the various sampling designs considered in this paper. The stratified sampling design can also provide more accurate results.

Keywords: poverty line, risk of poverty, auxiliary variable, ratio method

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Authors: Hallan Moura Ladeira, Carla Tatiana Mota Anflor

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The drilling process for bolted or riveted joints of components is very common in the naval, aeronautical, mechanical, and civil industries. In this context, the present work aims to study, through computer simulation, the influence of hole geometry (through, chamfered, and rounded) on crack propagation when submitted to static and dynamic loads. For the static crack evaluation, failure was considered when the stress intensity factor (FIT) exceeds the fracture toughness of the material (KIc). In the case of fatigue, the condition of the small crack tip plastification zone and the Paris Law were considered for determining region II of the dadN x ΔK curve. Initially, a parametric analysis of the hole geometry was performed to obtain a topology that would result in less discontinuity of the stress field and, consequently, less influence on static crack growth. The best performing topology was then used to study the fatigue crack growth rate considering the Paris Law. The numerical tests were performed on a 7075-T6 aluminum specimen resulting in dadN x ΔK curves in good agreement with the literature.

Keywords: holes, cracks, loading, fracture toughness

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Authors: Erika Tatiana Ruiz, Wilson Adarme Jaimes

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This abstract presents the methodology for improving the logistics processes of agricultural production units belonging to the coffee, cocoa, and fruit sectors, starting from the fundamental concepts and detailing each of the phases to carry out the diagnosis, which will be the basis for the formulation of its action plan and implementation of the maturity model. As a result of this work, the maturity model is formulated to improve logistics processes. This model seeks to: generate a progressive model that is useful for all productive units belonging to these sectors at the national level, regardless of their initial conditions, focus on the improvement of logistics processes as a strategy that contributes to improving the competitiveness of the agricultural sector in Colombia and spread the implementation of good logistics practices in postharvest in all departments of the country through autonomous tools. This model has been built through a series of steps that allow the evaluation and improvement of the logistics dimensions or indicators. The potential improvements for each dimension provide the foundation on which to advance to the next level. Within the maturity model, a methodology is indicated for the design and execution of strategies to improve its logistics processes, taking into account the current state of each production unit.

Keywords: agroindustrial, characterization, logistics, maturity model, processes

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Authors: Tatiana V. Raudina, Sergey P. Kulizhskiy

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A large part of Russia is located in permafrost areas. These areas are widely used because there are concentrated valuable natural resources. Therefore to explore of cryosols it is important due to the significant increase of anthropogenic stress as well as the problem of global climate change. In the north of Western Siberia permafrost phenomena is widespread. Permafrost as a factor of soil formation and cryogenesis as a process have a great impact on the soil formation of these areas. Based on the research results of permafrost-affected soils tundra landscapes formed in the central part of the Tazovskiy Peninsula in cryogenic conditions, data were obtained which characterize the morphological features of soils. The specificity of soil cover distribution and manifestation of soil-forming processes within the study area are noted. Permafrost features such as frost cracking, cryoturbation, thixotropy, movement of humus are formed. The formation of these features is increased with the development of the territory. As a consequence, there is a change in the components of the environment and the destruction of the soil cover.

Keywords: gleyed and nongleyed soils, permafrost, soil cryogenesis (pedocryogenesis), soil-forming macroprocesses

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Authors: Leonardo Alfredo Mendoza, Cristian Munoz, Marco Aurelio Pacheco, Manoela Kohler, Evelyn Batista, Rodrigo Moura

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Natural Language Processing (NLP) techniques are increasingly more powerful to be able to interpret the feelings and reactions of a person to a product or service. Sentiment analysis has become a fundamental tool for this interpretation but has few applications in languages other than English. This paper presents a classification of sentiment analysis in Portuguese with a base of comments from social networks in Portuguese. A word embedding's representation was used with a 50-Dimension GloVe pre-trained model, generated through a corpus completely in Portuguese. To generate this classification, the bidirectional long short-term memory and bidirectional Gated Recurrent Unit (GRU) models are used, reaching results of 99.1%.

Keywords: natural processing language, sentiment analysis, bidirectional long short-term memory, BI-LSTM, gated recurrent unit, GRU

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Authors: Tatiana R. Simonyan, Elena A. Protasova, Anastasia V. Mamontova, Eugene G. Maksimov, Konstantin A. Lukyanov, Alexey M. Bogdanov

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Here, we describe two variants of the calcium indicators based on the GCaMP sensitive core and BrUSLEE fluorescent protein (GCaMP-BrUSLEE and GCaMP-BrUSLEE-145). In contrast to the conventional GCaMP6-family indicators, these fluorophores are characterized by the well-marked responsiveness of their fluorescence decay kinetics to external calcium concentration both in vitro and in cellulo. Specifically, we show that the purified GCaMP-BrUSLEE and GCaMP-BrUSLEE-145 exhibit three-component fluorescence decay kinetics, with the amplitude-normalized lifetime component (t3*A3) of GCaMP-BrUSLEE-145 changing four-fold (500-2000 a.u.) in response to a Ca²⁺ concentration shift in the range of 0—350 nM. Time-resolved fluorescence microscopy of live cells displays the two-fold change of the GCaMP-BrUSLEE-145 mean lifetime upon histamine-stimulated calcium release. The aforementioned Ca²⁺-dependence calls considering the GCaMP-BrUSLEE-145 as a prospective Ca²⁺-indicator with the signal read-out in the time domain.

Keywords: calcium imaging, fluorescence lifetime imaging microscopy, fluorescent proteins, genetically encoded indicators

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