Search results for: Pascal Ortega
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 30

Search results for: Pascal Ortega

30 Infinite Impulse Response Digital Filters Design

Authors: Phuoc Si Nguyen

Abstract:

Infinite impulse response (IIR) filters can be designed from an analogue low pass prototype by using frequency transformation in the s-domain and bilinear z-transformation with pre-warping frequency; this method is known as frequency transformation from the s-domain to the z-domain. This paper will introduce a new method to transform an IIR digital filter to another type of IIR digital filter (low pass, high pass, band pass, band stop or narrow band) using a technique based on inverse bilinear z-transformation and inverse matrices. First, a matrix equation is derived from inverse bilinear z-transformation and Pascal’s triangle. This Low Pass Digital to Digital Filter Pascal Matrix Equation is used to transform a low pass digital filter to other digital filter types. From this equation and the inverse matrix, a Digital to Digital Filter Pascal Matrix Equation can be derived that is able to transform any IIR digital filter. This paper will also introduce some specific matrices to replace the inverse matrix, which is difficult to determine due to the larger size of the matrix in the current method. This will make computing and hand calculation easier when transforming from one IIR digital filter to another in the digital domain.

Keywords: bilinear z-transformation, frequency transformation, inverse bilinear z-transformation, IIR digital filters

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29 Effects of Voltage Pulse Characteristics on Some Performance Parameters of LiₓCoO₂-based Resistive Switching Memory Devices

Authors: Van Son Nguyen, Van Huy Mai, Alec Moradpour, Pascale Auban Senzier, Claude Pasquier, Kang Wang, Pierre-Antoine Albouy, Marcelo J. Rozenberg, John Giapintzakis, Christian N. Mihailescu, Charis M. Orfanidou, Thomas Maroutian, Philippe Lecoeur, Guillaume Agnus, Pascal Aubert, Sylvain Franger, Raphaël Salot, Nathalie Brun, Katia March, David Alamarguy, Pascal ChréTien, Olivier Schneegans

Abstract:

In the field of Nanoelectronics, a major research activity is being developed towards non-volatile memories. To face the limitations of existing Flash memory cells (endurance, downscaling, rapidity…), new approaches are emerging, among them resistive switching memories (Re-RAM). In this work, we analysed the behaviour of LixCoO2 oxide thin films in electrode/film/electrode devices. Preliminary results have been obtained concerning the influence of bias pulses characteristics (duration, value) on some performance parameters, such as endurance and resistance ratio (ROFF/RON). Besides, Conducting Probe Atomic Force Microscopy (CP-AFM) characterizations of the devices have been carried out to better understand some causes of performance failure, and thus help optimizing the switching performance of such devices.

Keywords: non volatile resistive memories, resistive switching, thin films, endurance

Procedia PDF Downloads 571
28 Photoreflectance Anisotropy Spectroscopy of Coupled Quantum Wells

Authors: J. V. Gonzalez Fernandez, T. Mozume, S. Gozu, A. Lastras Martinez, L. F. Lastras Martinez, J. Ortega Gallegos, R. E. Balderas Navarro

Abstract:

We report on a theoretical-experimental study of photoreflectance anisotropy (PRA) spectroscopy of coupled double quantum wells. By probing the in-plane interfacial optical anisotropies, we demonstrate that PRA spectroscopy has the capacity to detect and distinguish layers with quantum dimensions. In order to account for the experimental PRA spectra, we have used a theoretical model at k=0 based on a linear electro-optic effect through a piezoelectric shear strain.

Keywords: coupled double quantum well (CDQW), linear electro-optic (LEO) effect, photoreflectance anisotropy (PRA), piezoelectric shear strain

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27 Tetracycline as Chemosensor for Simultaneous Recognition of Al³⁺: Application to Bio-Imaging for Living Cells

Authors: Jesus Alfredo Ortega Granados, Pandiyan Thangarasu

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Antibiotic tetracycline presents as a micro-contaminant in fresh water, wastewater and soils, causing environmental and health problems. In this work, tetracycline (TC) has been employed as chemo-sensor for the recognition of Al³⁺ without interring other ions, and the results show that it enhances the fluorescence intensity for Al³⁺ and there is no interference from other coexisting cation ions (Cd²⁺, Ni²⁺, Co²⁺, Sr²⁺, Mg²⁺, Fe³⁺, K⁺, Sm³⁺, Ag⁺, Na⁺, Ba²⁺, Zn²⁺, and Mn²⁺). For the addition of Cu²⁺ to [TET-Al³⁺], it appears that the intensity of fluorescence has been quenched. Other combinations of metal ions in addition to TC do not change the fluorescence behavior. The stoichiometry determined by Job´s plot for the interaction of TC with Al³⁺ was found to be 1:1. Importantly, the detection of Al³⁺⁺ successfully employed in the real samples like living cells, and it was found that TC efficiently performs as a fluorescent probe for Al³⁺ ion in living systems, especially in Saccharomyces cerevisiae; this is confirmed by confocal laser scanning microscopy.

Keywords: chemo-sensor, recognition of Al³⁺ ion, Saccharomyces cerevisiae, tetracycline,

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26 A Comparative Study of Particle Image Velocimetry (PIV) and Particle Tracking Velocimetry (PTV) for Airflow Measurement

Authors: Sijie Fu, Pascal-Henry Biwolé, Christian Mathis

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Among modern airflow measurement methods, Particle Image Velocimetry (PIV) and Particle Tracking Velocimetry (PTV), as visualized and non-instructive measurement techniques, are playing more important role. This paper conducts a comparative experimental study for airflow measurement employing both techniques with the same condition. Velocity vector fields, velocity contour fields, voticity profiles and turbulence profiles are selected as the comparison indexes. The results show that the performance of both PIV and PTV techniques for airflow measurement is satisfied, but some differences between the both techniques are existed, it suggests that selecting the measurement technique should be based on a comprehensive consideration.

Keywords: airflow measurement, comparison, PIV, PTV

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25 Bag of Words Representation Based on Weighting Useful Visual Words

Authors: Fatma Abdedayem

Abstract:

The most effective and efficient methods in image categorization are almost based on bag-of-words (BOW) which presents image by a histogram of occurrence of visual words. In this paper, we propose a novel extension to this method. Firstly, we extract features in multi-scales by applying a color local descriptor named opponent-SIFT. Secondly, in order to represent image we use Spatial Pyramid Representation (SPR) and an extension to the BOW method which based on weighting visual words. Typically, the visual words are weighted during histogram assignment by computing the ratio of their occurrences in the image to the occurrences in the background. Finally, according to classical BOW retrieval framework, only a few words of the vocabulary is useful for image representation. Therefore, we select the useful weighted visual words that respect the threshold value. Experimentally, the algorithm is tested by using different image classes of PASCAL VOC 2007 and is compared against the classical bag-of-visual-words algorithm.

Keywords: BOW, useful visual words, weighted visual words, bag of visual words

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24 Developing an Effectual Logic through a Visual Mind Mapping

Authors: Alberti Pascal, Mustapha Mouloua

Abstract:

Companies are confronted with complex and competitive markets. The dynamics of these markets are becoming more and more fluid, requiring companies to provide competitive, definite and technological responses within increasingly short timeframes. To meet this demand, companies must rely on the cognitive abilities of actors of creativity to provide tangible answers to current contextual problems. It therefore seems appropriate to provide instruments to support this particular stage of innovation. Various methods and tools can meet this requirement. For a number of years we have been conducting experiments on the use of mind maps in the context of innovation projects with teams of different nationalities. After presenting the main research carried out on this theme, we discuss the possible correlation between the different uses of iconic tools and certain types of innovation. We then provide a link with different cognitive logic. Finally, we conclude by putting our research into perspective.

Keywords: creativity, innovation, causal logic, effectual logic, mind mapping

Procedia PDF Downloads 387
23 Lightweight High-Pressure Ratio Centrifugal Compressor for Vehicles-Investigation of Pipe Diffuser Designs by Means of CFD

Authors: Eleni Ioannou, Pascal Nucara, Keith Pullen

Abstract:

The subject of this paper is the investigation of the best efficiency design of a compressor diffuser applied in new lightweight, ultra efficient micro-gas turbine engines for vehicles. The Computational Fluid Dynamics (CFD) results are obtained utilizing steady state simulations for a wedge and an ”oval” type pipe diffuser in an effort to identify the beneficial effects of the pipe diffuser design. The basic flow features are presented with particular focus on the optimization of the pipe diffuser leading to higher efficiencies for the compressor stage. The optimised pipe diffuser is designed to exploit the 3D freedom enabled by Selective Laser Melting, hence purposely involves an investigation of geometric characteristics that do not follow the traditional diffuser concept.

Keywords: CFD, centrifugal compressor, micro-gas turbine, pipe diffuser, SLM, wedge diffuser

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22 Optimizing the Capacity of a Convolutional Neural Network for Image Segmentation and Pattern Recognition

Authors: Yalong Jiang, Zheru Chi

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In this paper, we study the factors which determine the capacity of a Convolutional Neural Network (CNN) model and propose the ways to evaluate and adjust the capacity of a CNN model for best matching to a specific pattern recognition task. Firstly, a scheme is proposed to adjust the number of independent functional units within a CNN model to make it be better fitted to a task. Secondly, the number of independent functional units in the capsule network is adjusted to fit it to the training dataset. Thirdly, a method based on Bayesian GAN is proposed to enrich the variances in the current dataset to increase its complexity. Experimental results on the PASCAL VOC 2010 Person Part dataset and the MNIST dataset show that, in both conventional CNN models and capsule networks, the number of independent functional units is an important factor that determines the capacity of a network model. By adjusting the number of functional units, the capacity of a model can better match the complexity of a dataset.

Keywords: CNN, convolutional neural network, capsule network, capacity optimization, character recognition, data augmentation, semantic segmentation

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21 Bag of Words Representation Based on Fusing Two Color Local Descriptors and Building Multiple Dictionaries

Authors: Fatma Abdedayem

Abstract:

We propose an extension to the famous method called Bag of words (BOW) which proved a successful role in the field of image categorization. Practically, this method based on representing image with visual words. In this work, firstly, we extract features from images using Spatial Pyramid Representation (SPR) and two dissimilar color descriptors which are opponent-SIFT and transformed-color-SIFT. Secondly, we fuse color local features by joining the two histograms coming from these descriptors. Thirdly, after collecting of all features, we generate multi-dictionaries coming from n random feature subsets that obtained by dividing all features into n random groups. Then, by using these dictionaries separately each image can be represented by n histograms which are lately concatenated horizontally and form the final histogram, that allows to combine Multiple Dictionaries (MDBoW). In the final step, in order to classify image we have applied Support Vector Machine (SVM) on the generated histograms. Experimentally, we have used two dissimilar image datasets in order to test our proposition: Caltech 256 and PASCAL VOC 2007.

Keywords: bag of words (BOW), color descriptors, multi-dictionaries, MDBoW

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20 Relationship Between Pain Intensity at the Time of the Hamstring Muscle Injury and Hamstring Muscle Lesion Volume Measured by Magnetic Resonance Imaging

Authors: Grange Sylvain, Plancher Ronan, Reurink Guustav, Croisille Pierre, Edouard Pascal

Abstract:

The primary objective of this study was to analyze the potential correlation between the pain experienced at the time of a hamstring muscle injury and the volume of the lesion measured on MRI. The secondary objectives were to analyze a correlation between this pain and the lesion grade as well as the affected hamstring muscle. We performed a retrospective analysis of the data collected in a prospective, multicenter, non-interventional cohort study (HAMMER). Patients with suspected hamstring muscle injury had an MRI after the injury and at the same time were evaluated for their pain intensity experienced at the time of the injury with a Numerical Pain Rating Scale (NPRS) from 0 to 10. A total of 61 patients were included in the present analysis. MRIs were performed in an average of less than 8 days. There was a significant correlation between pain and the injury volume (r=0.287; p=0.025). There was no significant correlation between the pain and the lesion grade (p>0.05), nor between the pain and affected hamstring muscle (p>0.05). Pain at the time of injury appeared to be correlated with the volume of muscle affected. These results confirm the value of a clinical approach in the initial evaluation of hamstring injuries to better select patients eligible for further imaging.

Keywords: hamstring muscle injury, MRI, volume lesion, pain

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19 A Contemporary Gender Predominance: A Honduran Textile Manufacturing Diagnose

Authors: Jesús David Argueta Moreno, Taria Ruiz, Cesar Ortega

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This qualitative investigation represents the first stage of the human capital engineering analysis, along the small and medium textile manufacturing companies, located on the city of Tegucigalpa, Honduras where the symptoms of the local manufacturing industry´s describe a severe gender displacement phenomenon. The evaluation of this phenomena, intends to trigger the Honduran small and medium technology manufactures into a collective performance, analysis through the development of a sectorial diagnose and the creation of a manufacturers guide, personalized. In accordance to the Honduran textile manufacturing needs, in order to strengthen their personnel capacities and thereby smoothen the gender equilibrium on this particular sector. It is worth mentioning, that on the last decade, the female gender has gathered positive statistics upon Central American job market´s, were the local business landscape describes a significant displacement of the Honduran female operators over the male gender workers that has significantly diminished their employment predominance. On the other hand, this study aims to evaluate the main features that impact on the job market local gender supplanting. On the other hand, this document aims to holistically describe the Honduran manufacturing context, as well as the current textile operator qualifications, in order to infer over the most proper human resources enforcement approaches/techniques on the industry.

Keywords: gender predominance, manufacturing, higher education institutions, emerging trends

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18 The Effect of Symmetrical Presentation of a "Photographic Mind Map" on the Production of Design Solutions

Authors: Pascal Alberti, Mustapha Mouloua

Abstract:

In today’s global market economy, various companies are often confronted with the dynamic and complex nature of current competitive markets. The dynamics of these markets are becoming more and more fluid, often requiring companies to provide competitive, definite advantages, and technological responses within increasingly shorte time frames. To meet these demands, companies must rely on the cognitive abilities of actors of creativity to provide tangible answers to the current contextual problems. Thus, it is important to provide a variety of instruments and design tools to support this particular stage of innovation, and to meet their demand expectations. For a number of years now, we have been extensively conducting experiments on the use of mind maps in the context of innovative projects with collaborative research teams from various nationalities. Our research findings reported a significant difference between a “Word” Mind Map and “Photographic” Mind Map, a correlation between the different uses of iconic tools and certain types of innovation, and a relationship between the different cognitive logics. In this paper, we will present our new results related to the effect of symmetrical presentation of a Photographic Mind Map" on the production of design solutions. Finally, we will conclude by highlighting the importance of our experimental method, and discussing both the theoretical and practical implications of our research.

Keywords: creativity, innovation, management, mind mapping, design product

Procedia PDF Downloads 477
17 Non-Targeted Adversarial Object Detection Attack: Fast Gradient Sign Method

Authors: Bandar Alahmadi, Manohar Mareboyana, Lethia Jackson

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Today, there are many applications that are using computer vision models, such as face recognition, image classification, and object detection. The accuracy of these models is very important for the performance of these applications. One challenge that facing the computer vision models is the adversarial examples attack. In computer vision, the adversarial example is an image that is intentionally designed to cause the machine learning model to misclassify it. One of very well-known method that is used to attack the Convolution Neural Network (CNN) is Fast Gradient Sign Method (FGSM). The goal of this method is to find the perturbation that can fool the CNN using the gradient of the cost function of CNN. In this paper, we introduce a novel model that can attack Regional-Convolution Neural Network (R-CNN) that use FGSM. We first extract the regions that are detected by R-CNN, and then we resize these regions into the size of regular images. Then, we find the best perturbation of the regions that can fool CNN using FGSM. Next, we add the resulted perturbation to the attacked region to get a new region image that looks similar to the original image to human eyes. Finally, we placed the regions back to the original image and test the R-CNN with the attacked images. Our model could drop the accuracy of the R-CNN when we tested with Pascal VOC 2012 dataset.

Keywords: adversarial examples, attack, computer vision, image processing

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16 Experimental Quantification and Modeling of Dissolved Gas during Hydrate Crystallization: CO₂ Hydrate Case

Authors: Amokrane Boufares, Elise Provost, Veronique Osswald, Pascal Clain, Anthony Delahaye, Laurence Fournaison, Didier Dalmazzone

Abstract:

Gas hydrates have long been considered as problematic for flow assurance in natural gas and oil transportation. On the other hand, they are now seen as future promising materials for various applications (i.e. desalination of seawater, natural gas and hydrogen storage, gas sequestration, gas combustion separation and cold storage and transport). Nonetheless, a better understanding of the crystallization mechanism of gas hydrate and of their formation kinetics is still needed for a better comprehension and control of the process. To that purpose, measuring the real-time evolution of the dissolved gas concentration in the aqueous phase during hydrate formation is required. In this work, CO₂ hydrates were formed in a stirred reactor equipped with an Attenuated Total Reflection (ATR) probe coupled to a Fourier Transform InfraRed (FTIR) spectroscopy analyzer. A method was first developed to continuously measure in-situ the CO₂ concentration in the liquid phase during solubilization, supersaturation, hydrate crystallization and dissociation steps. Thereafter, the measured concentration data were compared with those of equilibrium concentrations. It was observed that the equilibrium is instantly reached in the liquid phase due to the fast consumption of dissolved gas by the hydrate crystallization. Consequently, it was shown that hydrate crystallization kinetics is limited by the gas transfer at the gas-liquid interface. Finally, we noticed that the liquid-hydrate equilibrium during the hydrate crystallization is governed by the temperature of the experiment under the tested conditions.

Keywords: gas hydrate, dissolved gas, crystallization, infrared spectroscopy

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15 Influence of the Granular Mixture Properties on the Rheological Properties of Concrete: Yield Stress Determination Using Modified Chateau et al. Model

Authors: Rachid Zentar, Mokrane Bala, Pascal Boustingorry

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The prediction of the rheological behavior of concrete is at the center of current concerns of the concrete industry for different reasons. The shortage of good quality standard materials combined with variable properties of available materials imposes to improve existing models to take into account these variations at the design stage of concrete. The main reasons for improving the predictive models are, of course, saving time and cost at the design stage as well as to optimize concrete performances. In this study, we will highlight the different properties of the granular mixtures that affect the rheological properties of concrete. Our objective is to identify the intrinsic parameters of the aggregates which make it possible to predict the yield stress of concrete. The work was done using two typologies of grains: crushed and rolled aggregates. The experimental results have shown that the rheology of concrete is improved by increasing the packing density of the granular mixture using rolled aggregates. The experimental program realized allowed to model the yield stress of concrete by a modified model of Chateau et al. through a dimensionless parameter following Krieger-Dougherty law. The modelling confirms that the yield stress of concrete depends not only on the properties of cement paste but also on the packing density of the granular skeleton and the shape of grains.

Keywords: crushed aggregates, intrinsic viscosity, packing density, rolled aggregates, slump, yield stress of concrete

Procedia PDF Downloads 85
14 Qualitative Analysis of Current Child Custody Evaluation Practices

Authors: Carolyn J. Ortega, Stephen E. Berger

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The role of the custody evaluator is perhaps one of the most controversial and risky endeavors in clinical practice. Complaints filed with licensing boards regarding a child-custody evaluation constitute the second most common reason for such an event. Although the evaluator is expected to answer for the family-law court what is in the “best interest of the child,” there is a lack of clarity on how to establish this in any empirically validated manner. Hence, practitioners must contend with a nebulous framework in formulating their methodological procedures that inherently places them at risk in an already litigious context. This study sought to qualitatively investigate patterns of practice among doctoral practitioners conducting child custody evaluations in the area of Southern California. Ten psychologists were interviewed who devoted between 25 and 100% of their California private practice to custody work. All held Ph.D. degrees with a range of eight to 36 years of experience in custody work. Semi-structured interviews were used to investigate assessment practices, ensure adherence to guidelines, risk management, and qualities of evaluators. Forty-three Specific Themes were identified using Interpretive Phenomenological Analysis (IPA). Seven Higher Order Themes clustered on salient factors such as use of Ethics, Law, Guidelines; Parent Variables; Child Variables; Psychologist Variables; Testing; Literature; and Trends. Evaluators were aware of the ever-present reality of a licensure complaint and thus presented idiosyncratic descriptions of risk management considerations. Ambiguity about quantifying and validly tapping parenting abilities was also reviewed. Findings from this study suggested a high reliance on unstructured and observational methods in child custody practices.

Keywords: forensic psychology, psychological testing, assessment methodology, child custody

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13 Performance in the Delivery of Environmental Management Programs of the Local Government Unit of Malay, Aklan, Philippines

Authors: Tomas O. Ortega, Cecilia T. Reyes, Cecile O. Legaspi, Cylde G. Abayon, Anna Mae C. Relingo, Mary Eden M. Teruel

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A study was conducted to evaluate the performance in the delivery of environmental management programs of the local government of Malay, Aklan, Philippines. The samples were determined by adopting the Multi-Stage Random Probability Sampling technique. The 150 respondents were drawn from barangays with larger shares of the population based on the Philippine Statistical Authority’s Data on Census Population and Housing for the year 2015. The qualified sample respondents were selected using the Kish Grid. Female respondents were targeted for even numbered questionnaires while male respondents were targeted for odd numbers. The four major core concepts namely awareness, availment, satisfaction and need for action were used in measuring the rating of the respondents and presented in frequency and percentage distributions. The reasons for their response were likewise gathered. The study inferred that a large portion of the respondents was profoundly aware of the environmental management programs implemented by their local government unit especially the solid waste management and the clean-up programs/projects. Programs to control air pollution and waste water management obtained the least awareness ratings from the respondents. A high percentage of respondents had availed of environmental management programs, particularly solid waste management. Overall, majority of the respondents were satisfied with the environmental management programs rendered by the local government unit and therefore needs less action. It is recommended that the local government unit must strengthen air pollution control program. Appropriate action must be taken to support the people’s interest in this program most particularly to the individuals who burn their garbage. Seminars and training-workshops about appropriate waste disposal will most likely help settle this issue.

Keywords: availment, awareness, environmental management, need for action, satisfaction

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12 An Experimental Quantitative Case Study of Competency-Based Learning in Online Mathematics Education

Authors: Pascal Roubides

Abstract:

The presentation proposed herein describes a research case study of a hybrid application of the competency-based education model best exemplified by Western Governor’s University, within the general temporal confines of an accelerated (8-week) term of a College Algebra course at the author’s institution. A competency-based model was applied to an accelerated online College Algebra course, built as an Open Educational Resources (OER) course, seeking quantifiable evidence of any differences in the academic achievement of students enrolled in the competency-based course and the academic achievement of the current delivery of the same course. Competency-based learning has been gaining in support in recent times and the author’s institution has also been involved in its own efforts to design and develop courses based on this approach. However, it is unknown whether there had been any research conducted to quantify evidence of the effect of this approach against traditional approaches prior to the author’s case study. The research question sought to answer in this experimental quantitative study was whether the online College Algebra curriculum at the author’s institution delivered via an OER-based competency-based model can produce statistically significant improvement in retention and success rates against the current delivery of the same course. Results obtained in this study showed that there is no statistical difference in the retention rate of the two groups. However, there was a statistically significant difference found between the rates of successful completion of students in the experimental group versus those in the control group.

Keywords: competency-based learning, online mathematics, online math education, online courses

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11 Breast Cancer Incidence Estimation in Castilla-La Mancha (CLM) from Mortality and Survival Data

Authors: C. Romero, R. Ortega, P. Sánchez-Camacho, P. Aguilar, V. Segur, J. Ruiz, G. Gutiérrez

Abstract:

Introduction: Breast cancer is a leading cause of death in CLM. (2.8% of all deaths in women and 13,8% of deaths from tumors in womens). It is the most tumor incidence in CLM region with 26.1% from all tumours, except nonmelanoma skin (Cancer Incidence in Five Continents, Volume X, IARC). Cancer registries are a good information source to estimate cancer incidence, however the data are usually available with a lag which makes difficult their use for health managers. By contrast, mortality and survival statistics have less delay. In order to serve for resource planning and responding to this problem, a method is presented to estimate the incidence of mortality and survival data. Objectives: To estimate the incidence of breast cancer by age group in CLM in the period 1991-2013. Comparing the data obtained from the model with current incidence data. Sources: Annual number of women by single ages (National Statistics Institute). Annual number of deaths by all causes and breast cancer. (Mortality Registry CLM). The Breast cancer relative survival probability. (EUROCARE, Spanish registries data). Methods: A Weibull Parametric survival model from EUROCARE data is obtained. From the model of survival, the population and population data, Mortality and Incidence Analysis MODel (MIAMOD) regression model is obtained to estimate the incidence of cancer by age (1991-2013). Results: The resulting model is: Ix,t = Logit [const + age1*x + age2*x2 + coh1*(t – x) + coh2*(t-x)2] Where: Ix,t is the incidence at age x in the period (year) t; the value of the parameter estimates is: const (constant term in the model) = -7.03; age1 = 3.31; age2 = -1.10; coh1 = 0.61 and coh2 = -0.12. It is estimated that in 1991 were diagnosed in CLM 662 cases of breast cancer (81.51 per 100,000 women). An estimated 1,152 cases (112.41 per 100,000 women) were diagnosed in 2013, representing an increase of 40.7% in gross incidence rate (1.9% per year). The annual average increases in incidence by age were: 2.07% in women aged 25-44 years, 1.01% (45-54 years), 1.11% (55-64 years) and 1.24% (65-74 years). Cancer registries in Spain that send data to IARC declared 2003-2007 the average annual incidence rate of 98.6 cases per 100,000 women. Our model can obtain an incidence of 100.7 cases per 100,000 women. Conclusions: A sharp and steady increase in the incidence of breast cancer in the period 1991-2013 is observed. The increase was seen in all age groups considered, although it seems more pronounced in young women (25-44 years). With this method you can get a good estimation of the incidence.

Keywords: breast cancer, incidence, cancer registries, castilla-la mancha

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10 A First Step towards Automatic Evolutionary for Gas Lifts Allocation Optimization

Authors: Younis Elhaddad, Alfonso Ortega

Abstract:

Oil production by means of gas lift is a standard technique in oil production industry. To optimize the total amount of oil production in terms of the amount of gas injected is a key question in this domain. Different methods have been tested to propose a general methodology. Many of them apply well-known numerical methods. Some of them have taken into account the power of evolutionary approaches. Our goal is to provide the experts of the domain with a powerful automatic searching engine into which they can introduce their knowledge in a format close to the one used in their domain, and get solutions comprehensible in the same terms, as well. These proposals introduced in the genetic engine the most expressive formal models to represent the solutions to the problem. These algorithms have proven to be as effective as other genetic systems but more flexible and comfortable for the researcher although they usually require huge search spaces to justify their use due to the computational resources involved in the formal models. The first step to evaluate the viability of applying our approaches to this realm is to fully understand the domain and to select an instance of the problem (gas lift optimization) in which applying genetic approaches could seem promising. After analyzing the state of the art of this topic, we have decided to choose a previous work from the literature that faces the problem by means of numerical methods. This contribution includes details enough to be reproduced and complete data to be carefully analyzed. We have designed a classical, simple genetic algorithm just to try to get the same results and to understand the problem in depth. We could easily incorporate the well mathematical model, and the well data used by the authors and easily translate their mathematical model, to be numerically optimized, into a proper fitness function. We have analyzed the 100 curves they use in their experiment, similar results were observed, in addition, our system has automatically inferred an optimum total amount of injected gas for the field compatible with the addition of the optimum gas injected in each well by them. We have identified several constraints that could be interesting to incorporate to the optimization process but that could be difficult to numerically express. It could be interesting to automatically propose other mathematical models to fit both, individual well curves and also the behaviour of the complete field. All these facts and conclusions justify continuing exploring the viability of applying the approaches more sophisticated previously proposed by our research group.

Keywords: evolutionary automatic programming, gas lift, genetic algorithms, oil production

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9 Refractory Cardiac Arrest: Do We Go beyond, Do We Increase the Organ Donation Pool or Both?

Authors: Ortega Ivan, De La Plaza Edurne

Abstract:

Background: Spain and other European countries have implemented Uncontrolled Donation after Cardiac Death (uDCD) programs. After 15 years of experience in Spain, many things have changed. Recent evidence and technical breakthroughs achieved in resuscitation are relevant for uDCD programs and raise some ethical concerns related to these protocols. Aim: To rethink current uDCD programs in the light of recent evidence on available therapeutic procedures applicable to victims of out-of-hospital cardiac arrest (OHCA). To address the following question: What is the current standard of treatment owed to victims of OHCA before including them in an uDCD protocol? Materials and Methods: Review of the scientific and ethical literature related to both uDCD programs and innovative resuscitation techniques. Results: 1) The standard of treatment received and the chances of survival of victims of OHCA depend on whether they are classified as Non-Heart Beating Patients (NHBP) or Non-Heart-Beating-Donors (NHBD). 2) Recent studies suggest that NHBPs are likely to survive, with good quality of life, if one or more of the following interventions are performed while ongoing CPR -guided by suspected or known cause of OHCA- is maintained: a) direct access to a Cath Lab-H24 or/and to extra-corporeal life support (ECLS); b) transfer in induced hypothermia from the Emergency Medical Service (EMS) to the ICU; c) thrombolysis treatment; d) mobile extra-corporeal membrane oxygenation (mini ECMO) instituted as a bridge to ICU ECLS devices. 3) Victims of OHCA who cannot benefit from any of these therapies should be considered as NHBDs. Conclusion: Current uDCD protocols do not take into account recent improvements in resuscitation and need to be adapted. Operational criteria to distinguish NHBDs from NHBP should seek a balance between the technical imperative (to do whatever is possible), considerations about expected survival with quality of life, and distributive justice (costs/benefits). Uncontrolled DCD protocols can be performed in a way that does not hamper the legitimate interests of patients, potential organ donors, their families, the organ recipients, and the health professionals involved in these processes. Families of NHBDs’ should receive information which conforms to the ethical principles of respect of autonomy and transparency.

Keywords: uncontrolled donation after cardiac death resuscitation, refractory cardiac arrest, out of hospital cardiac, arrest ethics

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8 Extraction and Quantification of Peramine Present in Dalaca pallens, a Pest of Grassland in Southtern Chile

Authors: Leonardo Parra, Daniel Martínez, Jorge Pizarro, Fernando Ortega, Manuel Chacón-Fuentes, Andrés Quiroz

Abstract:

Control of Dalaca pallens or blackworms, one of the most important hypogeous pest in grassland in southern Chile, is based on the use of broad-spectrum insecticides such as organophosphates and pyrethroids. However, the rapid development of insecticide resistance in field populations of this insect and public concern over the environmental impact of these insecticides has resulted in the search for other control methods. Specifically, the use of endophyte fungi for controlling pest has emerged as an interesting and promising strategy. Endophytes from ryegrass (Lolium perenne), establish a biotrophic relationship with the host, defined as mutualistic symbiosis. The plant-fungi association produces alkaloids where peramine is the main toxic substance against Listronotus bonariensis, the most important epigean pest of ryegrass. Nevertheless, the effect of peramina on others pest insects, such as D. pallens, to our knowledge has not been studied, and also its possible metabolization in the body of the larvae. Therefore, we addressed the following research question: Do larvae of D. pallens store peramine after consumption of ryegrass endophyte infected (E+)? For this, specimens of blackworms were fed with ryegrass plant of seven experimental lines and one commercial cultivar endophyte free (E-) sown at the Instituto de Investigaciones Agropecuarias Carillanca (Vilcún, Chile). Once the feeding period was over, ten larvae of each treatment were examined. Individuals were dissected, and their gut was removed to exclude any influence of remaining material. The rest of the larva's body was dried at 60°C by 24-48 h and ground into a fine powder using a mortar. 25 mg of dry powder was transferred to a microcentrifuge tube and extracted in 1 mL of a mixture of methanol:water:formic acid. Then, the samples were centrifuged at 16,000 rpm for 3 min, and the supernatant was colected and injected in the liquid chromatography of high resolution (HPLC). The results confirmed the presence of peramine in the larva's body of D. pallens. The insects that fed the experimental lines LQE-2 and LQE-6 were those where peramine was present in high proportion (0.205 and 0.199 ppm, respectively); while LQE-7 and LQE-3 obtained the lowest concentrations of the alkaloid (0.047 and 0.053 ppm, respectively). Peramine was not detected in the insects when the control cultivar Jumbo (E-) was tested. These results evidenced the storage and metabolism of peramine during consumption of the larvae. However, the effect of this alkaloid present in 'future ryegrass cultivars' (LQE-2 and LQE-6) on the performance and survival of blackworms must be studied and confirmed experimentally.

Keywords: blackworms, HPLC, alkaloid, pest

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7 Effect of Fermented Orange Juice Intake on Urinary 6‑Sulfatoxymelatonin in Healthy Volunteers

Authors: I. Cerrillo, A. Carrillo-Vico, M. A. Ortega, B. Escudero-López, N. Álvarez-Sánchez, F. Martín, M. S. Fernández-Pachón

Abstract:

Melatonin is a bioactive compound involved in multiple biological activities such as glucose tolerance, circadian rhythm regulation, antioxidant defense or immune system action. In elderly subjects the intake of foods and drinks rich in melatonin is very important due to its endogenous level decreases with age. Alcoholic fermentation is a process carried out in fruits, vegetables and legumes to obtain new products with improved bioactive compounds profile in relation to original substrates. Alcoholic fermentation process carried out by Saccharomycetaceae var. Pichia kluyveri induces an important synthesis of melatonin in orange juice. A novel beverage derived of fermented orange juice could be a promising source of this bioactive compound. The aim of the present study was to determine whether the acute intake of fermented orange juice increase the levels of urinary 6-sulfatoxymelatonin in healthy humans. Nine healthy volunteers (7 women and 2 men), aged between 20 and 25 years old and BMI of 21.1  2.4 kg/m2, were recruited. On the study day, participants ingested 500 mL of fermented orange juice. The first urine collection was made before fermented orange juice consumption (basal). The rest of urine collections were made in the following time intervals after fermented orange juice consumption: 0-2, 2-5, 5-10, 10- 15 and 15-24 hours. During the experimental period only the consumption of water was allowed. At lunch time a meal was provided (60 g of white bread, two slices of ham, a slice of cheese, 125 g of sweetened natural yoghurt and water). The subjects repeated the protocol with orange juice following a 2-wk washout period between both types of beverages. The levels of 6-sulfatoxymelatonin (6-SMT) were measured in urine recollected at different time points using the Melatonin-Sulfate Urine ELISA (IBL International GMBH, Hamburg, Germany). Levels of 6-SMT were corrected to those of creatinine for each sample. A significant (p < 0.05) increase in urinary 6-SMT levels was observed between 2-5 hours after fermented orange juice ingestion with respect to basal values (increase of 67,8 %). The consumption of orange juice did not induce any significant change in urinary 6-SMT levels. In addition, urinary 6-SMT levels obtained between 2-5 hours after fermented orange juice ingestion (115,6 ng/mg) were significantly different (p < 0.05) from those of orange juice (42,4 ng/mg). The enhancement of urinary 6-SMT after the ingestion of 500 mL of fermented orange juice in healthy humans compared to orange juice could be an important advantage of this novel product as an excellent source of melatonin. Fermented orange juice could be a new functional food, and its consumption could exert a potentially positive effect on health in both the maintenance of health status and the prevention of chronic diseases.

Keywords: fermented orange juice, functional beverage, healthy human, melatonin

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6 Production, Characterization and In vitro Evaluation of [223Ra]RaCl2 Nanomicelles for Targeted Alpha Therapy of Osteosarcoma

Authors: Yang Yang, Luciana Magalhães Rebelo Alencar, Martha Sahylí Ortega Pijeira, Beatriz da Silva Batista, Alefe Roger Silva França, Erick Rafael Dias Rates, Ruana Cardoso Lima, Sara Gemini-Piperni, Ralph Santos-Oliveira

Abstract:

Radium-²²³ dichloride ([²²³Rₐ]RₐCl₂) is an alpha particle-emitting radiopharmaceutical currently approved for the treatment of patients with castration-resistant prostate cancer, symptomatic bone metastases, and no known visceral metastatic disease. [²²³Rₐ]RₐCl₂ is bone-seeking calcium mimetic that bonds into the newly formed bone stroma, especially osteoblastic or sclerotic metastases, killing the tumor cells by inducing DNA breaks in a potent and localized manner. Nonetheless, the successful therapy of osteosarcoma as primary bone tumors is still a challenge. Nanomicelles are colloidal nanosystems widely used in drug development to improve blood circulation time, bioavailability, and specificity of therapeutic agents, among other applications. In addition, the enhanced permeability and retention effect of the nanosystems, and the renal excretion of the nanomicelles reported in most cases so far, are very attractive to achieve selective and increased accumulation in tumor site as well as to increase the safety of [²²³Rₐ]RₐCl₂ in the clinical routine. In the present work, [²²³Rₐ]RₐCl₂ nanomicelles were produced, characterized, in vitro evaluated, and compared with pure [²²³Rₐ]RₐCl2 solution using SAOS2 osteosarcoma cells. The [²²³Rₐ]RₐCl₂ nanomicelles were prepared using the amphiphilic copolymer Pluronic F127. The dynamic light scattering analysis of freshly produced [²²³Rₐ]RₐCl₂ nanomicelles demonstrated a mean size of 129.4 nm with a polydispersity index (PDI) of 0.303. After one week stored in the refrigerator, the mean size of the [²²³Rₐ]RₐCl₂ nanomicelles increased to 169.4 with a PDI of 0.381. Atomic force microscopy analysis of [223Rₐ]RₐCl₂ nanomicelles exhibited spherical structures whose heights reach 1 µm, suggesting the filling of 127-Pluronic nanomicelles with [²²³Rₐ]RₐCl₂. The viability assay with [²²³Rₐ]RₐCl₂ nanomicelles displayed a dose-dependent response as it was observed using pure [²²³Rₐ]RₐCl2. However, at the same dose, [²²³Rₐ]RₐCl₂ nanomicelles were 20% higher efficient in killing SAOS2 cells when compared with pure [²²³Rₐ]RₐCl₂. These findings demonstrated the effectiveness of the nanosystem validating the application of nanotechnology in targeted alpha therapy with [²²³Ra]RₐCl₂. In addition, the [²²³Rₐ]RaCl₂nanomicelles may be decorated and incorporated with a great variety of agents and compounds (e.g., monoclonal antibodies, aptamers, peptides) to overcome the limited use of [²²³Ra]RₐCl₂.

Keywords: nanomicelles, osteosarcoma, radium dichloride, targeted alpha therapy

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5 Motivational Profiles of the Entrepreneurial Career in Spanish Businessmen

Authors: Magdalena Suárez-Ortega, M. Fe. Sánchez-García

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This paper focuses on the analysis of the motivations that lead people to undertake and consolidate their business. It is addressed from the framework of planned behavior theory, which recognizes the importance of the social environment and cultural values, both in the decision to undertake business and in business consolidation. Similarly, it is also based on theories of career development, which emphasize the importance of career management competencies and their connections to other vital aspects of people, including their roles within their families and other personal activities. This connects directly with the impact of entrepreneurship on the career and the professional-personal project of each individual. This study is part of the project titled Career Design and Talent Management (Ministry of Economy and Competitiveness of Spain, State Plan 2013-2016 Excellence Ref. EDU2013-45704-P). The aim of the study is to identify and describe entrepreneurial competencies and motivational profiles in a sample of 248 Spanish entrepreneurs, considering the consolidated profile and the profile in transition (n = 248).In order to obtain the information, the Questionnaire of Motivation and conditioners of the entrepreneurial career (MCEC) has been applied. This consists of 67 items and includes four scales (E1-Conflicts in conciliation, E2-Satisfaction in the career path, E3-Motivations to undertake, E4-Guidance Needs). Cluster analysis (mixed method, combining k-means clustering with a hierarchical method) was carried out, characterizing the groups profiles according to the categorical variables (chi square, p = 0.05), and the quantitative variables (ANOVA). The results have allowed us to characterize three motivational profiles relevant to the motivation, the degree of conciliation between personal and professional life, and the degree of conflict in conciliation, levels of career satisfaction and orientation needs (in the entrepreneurial project and life-career). The first profile is formed by extrinsically motivated entrepreneurs, professionally satisfied and without conflict of vital roles. The second profile acts with intrinsic motivation and also associated with family models, and although it shows satisfaction with their professional career, it finds a high conflict in their family and professional life. The third is composed of entrepreneurs with high extrinsic motivation, professional dissatisfaction and at the same time, feel the conflict in their professional life by the effect of personal roles. Ultimately, the analysis has allowed us to line the kinds of entrepreneurs to different levels of motivation, satisfaction, needs and articulation in professional and personal life, showing characterizations associated with the use of time for leisure, and the care of the family. Associations related to gender, age, activity sector, environment (rural, urban, virtual), and the use of time for domestic tasks are not identified. The model obtained and its implications for the design of training actions and orientation to entrepreneurs is also discussed.

Keywords: motivation, entrepreneurial career, guidance needs, life-work balance, job satisfaction, assessment

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4 A Strategy to Reduce Salt Intake: The Use of a Seasoning Obtained from Wine Pomace

Authors: María Luisa Gonzalez-SanJose, Javier Garcia-Lomillo, Raquel Del Pino, Miriam Ortega-Heras, Maria Dolores Rivero-Perez, Pilar Muñiz-Rodriguez

Abstract:

One of the most preoccupant problems related to the diet of the occidental societies is the high salt intake. In Spain, salt intake is almost twice as recommended by the World Health Organization (WHO). A lot of negative health effects of high sodium intake have been described being the hypertension, cardiovascular and coronary diseases ones of the most important. Due to this fact, government and other institutions are working on the gradual reduction of this consumption. Intake of meat products have been described as the main processed products that bring salt to the diet, followed by snacks and savory crackers. However, fortunately, the food industry has also raised awareness of this problem and is working intensely, and in recent years attempts to reduce the salt content in processed products, and is developing special lines with low sodium content. It is important to consider that processed food are the main source of sodium in occidental countries. One of the possible strategies to reduce the salt content in food is to find substitutes that can emulate their taste properties without adding much sodium or products that mask or substitute salty sensations with other flavors and aromas. In this sense, multiple products have been proposed and used until now. Potassium salts produce similar salty sensations without bring sodium, however their intake should be also limited, by healthy reasons. Furthermore, some potassium salts shows some better notes. Other alternatives are the use of flavor enhancers, spices, aromatic herbs, sea-plant derivate products, etc. The wine pomace is rich in potassium salts, content organic acid and other flavored substances, therefore it could be an interesting raw material to obtain derived products that could be useful as alternative ‘seasonings’. Considering previous comments, the main aim of this study was to evaluate the possible use of a natural seasoning, made from red wine pomace, in two different foods, crackers and burgers. The seasoning was made in the pilot plant of food technology of the University of Burgos, where the studied crackers and patties were also made. Different members of the University, students, docent and administrative personal, taste the products, and a trained panel evaluated salty intensity. The seasoning in addition to potassium contain significant levels of dietary fiber and phenolic compounds, which also makes it interesting as a functional ingredient. Both burgers and crackers made with the seasoning showed better taste that those without salt. Obviously, they showed lower sodium content than normal formulation, and were richer in potassium, antioxidant and fiber. Then, they showed lower values of the relation Na/K. All these facts are correlated with more ‘healthy’ products especially to that people with hypertension and other coronary dysfunctions.

Keywords: healthy foods, low salt, seasoning, wine pomace

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3 Management of Caverno-Venous Leakage: A Series of 133 Patients with Symptoms, Hemodynamic Workup, and Results of Surgery

Authors: Allaire Eric, Hauet Pascal, Floresco Jean, Beley Sebastien, Sussman Helene, Virag Ronald

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Background: Caverno-venous leakage (CVL) is devastating, although barely known disease, the first cause of major physical impairment in men under 25, and responsible for 50% of resistances to phosphodiesterase 5-inhibitors (PDE5-I), affecting 30 to 40% of users in this medication class. In this condition, too early blood drainage from corpora cavernosa prevents penile rigidity and penetration during sexual intercourse. The role of conservative surgery in this disease remains controversial. Aim: Assess complications and results of combined open surgery and embolization for CVL. Method: Between June 2016 and September 2021, 133 consecutive patients underwent surgery in our institution for CVL, causing severe erectile dysfunction (ED) resistance to oral medical treatment. Procedures combined vein embolization and ligation with microsurgical techniques. We performed a pre-and post-operative clinical (Erection Harness Scale: EHS) hemodynamic evaluation by duplex sonography in all patients. Before surgery, the CVL network was visualized by computed tomography cavernography. Penile EMG was performed in case of diabetes or suspected other neurological conditions. All patients were optimized for hormonal status—data we prospectively recorded. Results: Clinical signs suggesting CVL were ED since age lower than 25, loss of erection when changing position, penile rigidity varying according to the position. Main complications were minor pulmonary embolism in 2 patients, one after airline travel, one with Factor V Leiden heterozygote mutation, one infection and three hematomas requiring reoperation, one decreased gland sensitivity lasting for more than one year. Mean pre-operative pharmacologic EHS was 2.37+/-0.64, mean pharmacologic post-operative EHS was 3.21+/-0.60, p<0.0001 (paired t-test). The mean EHS variation was 0.87+/-0.74. After surgery, 81.5% of patients had a pharmacologic EHS equal to or over 3, allowing for intercourse with penetration. Three patients (2.2%) experienced lower post-operative EHS. The main cause of failure was leakage from the deep dorsal aspect of the corpus cavernosa. In a 14 months follow-up, 83.2% of patients had a clinical EHS equal to or over 3, allowing for sexual intercourse with penetration, one-third of them without any medication. 5 patients had a penile implant after unsuccessful conservative surgery. Conclusion: Open surgery combined with embolization for CVL is an efficient approach to CVL causing severe erectile dysfunction.

Keywords: erectile dysfunction, cavernovenous leakage, surgery, embolization, treatment, result, complications, penile duplex sonography

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2 Isoflavonoid Dynamic Variation in Red Clover Genotypes

Authors: Andrés Quiroz, Emilio Hormazábal, Ana Mutis, Fernando Ortega, Loreto Méndez, Leonardo Parra

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Red clover root borer, Hylastinus obscurus Marsham (Coleoptera: Curculionidae), is the main insect pest associated to red clover, Trifolium pratense L. An average of 1.5 H. obscurus per plant can cause 5.5% reduction in forage yield in pastures of two to three years old. Moreover, insect attack can reach 70% to 100% of the plants. To our knowledge, there is no a chemical strategy for controlling this pest. Therefore alternative strategies for controlling H. obscurus are a high priority for red clover producers. One of this alternative is related to the study of secondary metabolites involved in intrinsic chemical defenses developed by plants, such as isoflavonoids. The isoflavonoids formononetin and daidzein have elicited an antifeedant and phagostimult effect on H. obscurus respectively. However, we do not know how is the dynamic variation of these isoflavonoids under field conditions. The main objective of this work was to evaluate the variation of the antifeedant isoflavonoids formononetin, the phagostimulant isoflavonoids daidzein, and their respective glycosides over time in different ecotypes of red clover. Fourteen red clover ecotypes (8 cultivars and 6 experimental lines), were collected at INIA-Carillanca (La Araucanía, Chile). These plants were established in October 2015 under irrigated conditions. The cultivars were distributed in a randomized complete block with three replicates. The whole plants were sampled in four times: 15th October 2016, 12th December 2016, 27th January 2017 and 16th March 2017 with sufficient amount of soil to avoid root damage. A polar fraction of isoflavonoid was obtained from 20 mg of lyophilized root tissue extracted with 2 mL of 80% MeOH for 16 h using an orbital shaker in the dark at room temperature. After, an aliquot of 1.4 mL of the supernatant was evaporated, and the residue was resuspended in 300 µL of 45% MeOH. The identification and quantification of isoflavonoid root extracts were performed by the injection of 20 µL into a Shimadzu HPLC equipped with a C-18 column. The sample was eluted with a mobile phase composed of AcOH: H₂O (1:9 v/v) as solvent A and CH₃CN as solvent B. The detection was performed at 260 nm. The results showed that the amount of aglycones was higher than the respective glycosides. This result is according to the biosynthetic pathway of flavonoids, where the formation of glycoside is further to the glycosides biosynthesis. The amount of formononetin was higher than daidzein. In roots, where H. obscurus spent the most part of its live cycle, the highest content of formononetin was found in G 27, Pawera, Sabtoron High, Redqueli-INIA and Superqueli-INIA cvs. (2.1, 1.8, 1.8, 1.6 and 1.0 mg g⁻¹ respectively); and the lowest amount of daidzein were found Superqueli-INIA (0.32 mg g⁻¹) and in the experimental line Sel Syn Int4 (0.24 mg g⁻¹). This ecotype showed a high content of formononetin (0.9 mg g⁻¹). This information, associated with cultural practices, could help farmers and breeders to reduce H. obscurus in grassland, selecting ecotypes with high content of formononetin and low amount of daidzein in the roots of red clover plants. Acknowledgements: FONDECYT 1141245 and 11130715.

Keywords: daidzein, formononetin, isoflavonoid glycosides, trifolium pratense

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1 Enzymatic Determination of Limonene in Red Clover Genotypes

Authors: Andrés Quiroz, Emilio Hormazabal, Ana Mutis, Fernando Ortega, Manuel Chacón-Fuentes, Leonardo Parra

Abstract:

Red clover (Trifolium pratense L.) is an important forage species in temperate regions of the world. The main limitation of this species worldwide is a lack of persistence related to the high mortality of plants due to a complex of biotic and abiotic factors, determining a life span of two or three seasons. Because of the importance of red clover in Chile, a red clover breeding program was started at INIA Carillanca Research Center in 1989, with the main objective of improving the survival of plants, forage yield, and persistence. The main selection criteria for selecting new varieties have been based on agronomical parameters and biotic factors. The main biotic factor associated with red clover mortality in Chile is Hylastinus obscurus (Coleoptera: Curculionidae). Both larval and adults feed on the roots, causing weakening and subsequent death of clover plants. Pesticides have not been successful for controlling infestations of this root borer. Therefore, alternative strategies for controlling this pest are a high priority for red clover producers. Currently, the role of semiochemical in the interaction between H. obscurus and red clover plants has been widely studied for our group. Specifically, from the red clover foliage has been identified limonene is eliciting repellency from the root borer. Limonene is generated in the plant from two independent biosynthetic pathways, the mevalonic acid, and deoxyxylulose pathway. Mevalonate pathway enzymes are localized in the cytosol, whereas the deoxyxylulose phosphate pathway enzymes are found in plastids. In summary, limonene can be determinated by enzymatic bioassay using GPP as substrate and by limonene synthase expression. Therefore, the main objective of this work was to study genetic variation of limonene in material provided by INIA´s Red Clover breeding program. Protein extraction was carried out homogenizing 250 mg of leave tissue and suspended in 6 mL of extraction buffer (PEG 1500, PVP-30, 20 mM MgCl2 and antioxidants) and stirred on ice for 20 min. After centrifugation, aliquots of 2.5 mL were desalted on PD-10 columns, resulting in a final volume of 3.5 mL. Protein determination was performed according to Bradford with BSA as a standard. Monoterpene synthase assays were performed with 50 µL of protein extracts transferred into gas-tight 2 mL crimp seal vials after addition of 4 µL MgCl₂ and 41 µL assay buffer. The assay was started by adding 5 µL of a GPP solution. The mixture was incubated for 30 min at 40 °C. Biosynthesized limonene was quantified in a GC equipped with a chiral column and using synthetic R and S-limonene standards. The enzymatic the production of R and S-limonene from different Superqueli-Carillanca genotypes is shown in this work. Preliminary results showed significant differences in limonene content among the genotypes analyzed. These results constitute an important base for selecting genotypes with a high content of this repellent monoterpene towards H. obscurus.

Keywords: head space, limonene enzymatic determination, red clover, Hylastinus obscurus

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