Search results for: Esteban Alejandro Armas Vega
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 149

Search results for: Esteban Alejandro Armas Vega

29 Experimental Design in Extraction of Pseudomonas sp. Protease from Fermented Broth by Polyethylene Glycol/Citrate Aqueous Two-Phase System

Authors: Omar Pillaca-Pullo, Arturo Alejandro-Paredes, Carol Flores-Fernandez, Marijuly Sayuri Kina, Amparo Iris Zavaleta

Abstract:

Aqueous two-phase system (ATPS) is an interesting alternative for separating industrial enzymes due to it is easy to scale-up and low cost. Polyethylene glycol (PEG) mixed with potassium phosphate or magnesium sulfate is one of the most frequently polymer/salt ATPS used, but the consequences of its use is a high concentration of phosphates and sulfates in wastewater causing environmental issues. Citrate could replace these inorganic salts due to it is biodegradable and does not produce toxic compounds. On the other hand, statistical design of experiments is widely used for ATPS optimization and it allows to study the effects of the involved variables in the purification, and to estimate their significant effects on selected responses and interactions. The 24 factorial design with four central points (20 experiments) was employed to study the partition and purification of proteases produced by Pseudomonas sp. in PEG/citrate ATPS system. ATPS was prepared with different sodium citrate concentrations [14, 16 and 18% (w/w)], pH values (7, 8 and 9), PEG molecular weight (2,000; 4,000 and 6,000 g/mol) and PEG concentrations [18, 20 and 22 % (w/w)]. All system components were mixed with 15% (w/w) of the fermented broth and deionized water was added to a final weight of 12.5 g. Then, the systems were mixed and kept at room temperature until to reach two-phases separation. Volumes of the top and bottom phases were measured, and aliquots from both phases were collected for subsequent proteolytic activity and total protein determination. Influence of variables such as PEG molar mass (MPEG), PEG concentration (CPEG), citrate concentration (CSal) and pH were evaluated on the following responses: purification factor (PF), activity yield (Y), partition coefficient (K) and selectivity (S). STATISTICA program version 10 was used for the analysis. According to the obtained results, higher levels of CPEG and MPEG had a positive effect on extraction, while pH did not influence on the process. On the other hand, the CSal could be related with low values of Y because of the citrate ions have a negative effect on solubility and enzymatic structure. The optimum values of Y (66.4 %), PF (1.8), K (5.5) and S (4.3) were obtained at CSal (18%), MPEG (6,000 g/mol), CPEG (22%) and pH 9. These results indicated that the PEG/citrate system is accurate to purify these Pseudomonas sp. proteases from fermented broth as a first purification step.

Keywords: citrate, polyethylene glycol, protease, Pseudomonas sp

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28 The Effect of Emotional Stimuli Related to Body Imbalance in Postural Control and the Phenomenological Experience of Young Healthy Adults

Authors: David Martinez-Pernia, Alvaro Rivera-Rei, Alejandro Troncoso, Gonzalo Forno, Andrea Slachevsky, David Huepe, Victoria Silva-Mack, Jorge Calderon, Mayte Vergara, Valentina Carrera

Abstract:

Background: Recent theories in the field of emotions have taken the relevance of motor control beyond a system related to personal autonomy (walking, running, grooming), and integrate it into the emotional dimension. However, to our best knowledge, there are no studies that specifically investigate how emotional stimuli related to motor control modify emotional states in terms of postural control and phenomenological experience. Objective: The main aim of this work is to investigate the emotions produced by stimuli of bodily imbalance (neutral, pleasant and unpleasant) in the postural control and the phenomenological experience of young, healthy adults. Methodology: 46 healthy young people are shown emotional videos (neutral, pleasant, motor unpleasant, and non-motor unpleasant) related to the body imbalance. During the period of stimulation of each of the videos (60 seconds) the participant is standing on a force platform to collect temporal and spatial data of postural control. In addition, the electrophysiological activity of the heart and electrodermal activity is recorded. In relation to the two unpleasant conditions (motor versus non-motor), a phenomenological interview is carried out to collect the subjective experience of emotion and body perception. Results: Pleasant and unpleasant emotional videos have significant changes with respect to the neutral condition in terms of greater area, higher mean velocity, and greater mean frequency power on the anterior-posterior axis. The results obtained with respect to the electrodermal response was that the pleasurable and unpleasant conditions produced a significant increase in the phasic component with respect to the neutral condition. Regarding the electrophysiology of the heart, no significant change was found in any condition. Phenomenological experiences in the two unpleasant conditions differ in body perception and the emotional meaning of the experience. Conclusion: Emotional stimuli related to bodily imbalance produce changes in postural control, electrodermal activity, and phenomenological experience. This experimental setting could be relevant to be implemented in people with motor disorders (Parkinson, Stroke, TBI) to know how emotions affect motor control.

Keywords: body imbalance stimuli, emotion, phenomenological experience, postural control

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27 Development of Polylactic Acid Insert with a Cinnamaldehyde-Betacyclodextrin Complex for Cape Gooseberry (Physalis Peruviana L.) Packed

Authors: Gómez S. Jennifer, Méndez V. Camila, Moncayo M. Diana, Vega M. Lizeth

Abstract:

The cape gooseberry is a climacteric fruit; Colombia is one of the principal exporters in the world. The environmental condition of temperature and relative moisture decreases the titratable acidity and pH. These conditions and fruit maturation result in the fungal proliferation of Botrytis cinerea disease. Plastic packaging for fresh cape gooseberries was used for mechanical damage protection but created a suitable atmosphere for fungal growth. Beta-cyclodextrins are currently implemented as coatings for the encapsulation of hydrophobic compounds, for example, with bioactive compounds from essential oils such as cinnamaldehyde, which has a high antimicrobial capacity. However, it is a volatile substance. In this article, the casting method was used to obtain a polylactic acid (PLA) polymer film containing the beta-cyclodextrin-cinnamaldehyde inclusion complex, generating an insert that allowed the controlled release of the antifungal substance in packed cape gooseberries to decrease contamination by Botrytis cinerea in a latent state during storage. For the encapsulation technique, three ratios for the cinnamaldehyde: beta-cyclodextrin inclusion complex were proposed: (25:75), (40:60), and (50:50). Spectrophotometry, colorimetry in L*a*b* coordinate space and scanning electron microscopy (SEM) were made for the complex characterization. Subsequently, two ratios of tween and water (40:60) and (50:50) were used to obtain the polylactic acid (PLA) film. To determine mechanical and physical parameters of colourimetry in L*a*b* coordinate space, atomic force microscopy and stereoscopy were done to determine the transparency and flexibility of the film; for both cases, Statgraphics software was used to determine the best ratio in each of the proposed phases, where for encapsulation it was (50:50) with an encapsulation efficiency of 65,92%, and for casting the ratio (40:60) obtained greater transparency and flexibility that permitted its incorporation into the polymeric packaging. A liberation assay was also developed under ambient temperature conditions to evaluate the concentration of cinnamaldehyde inside the packaging through gas chromatography for three weeks. It was found that the insert had a controlled release. Nevertheless, a higher cinnamaldehyde concentration is needed to obtain the minimum inhibitory concentration for the fungus Botrytis cinerea (0.2g/L). The homogeneity of the cinnamaldehyde gas phase inside the packaging can be improved by considering other insert configurations. This development aims to impact emerging food preservation technologies with the controlled release of antifungals to reduce the affectation of the physico-chemical and sensory properties of the fruit as a result of contamination by microorganisms in the postharvest stage.

Keywords: antifungal, casting, encapsulation, postharvest

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26 Modelling of Air-Cooled Adiabatic Membrane-Based Absorber for Absorption Chillers Using Low Temperature Solar Heat

Authors: M. Venegas, M. De Vega, N. García-Hernando

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Absorption cooling chillers have received growing attention over the past few decades as they allow the use of low-grade heat to produce the cooling effect. The combination of this technology with solar thermal energy in the summer period can reduce the electricity consumption peak due to air-conditioning. One of the main components, the absorber, is designed for simultaneous heat and mass transfer. Usually, shell and tubes heat exchangers are used, which are large and heavy. Cooling water from a cooling tower is conventionally used to extract the heat released during the absorption and condensation processes. These are clear inconvenient for the generalization of the absorption technology use, limiting its benefits in the contribution to the reduction in CO2 emissions, particularly for the H2O-LiBr solution which can work with low heat temperature sources as provided by solar panels. In the present work a promising new technology is under study, consisting in the use of membrane contactors in adiabatic microchannel mass exchangers. The configuration here proposed consists in one or several modules (depending on the cooling capacity of the chiller) that contain two vapour channels, separated from the solution by adjacent microporous membranes. The solution is confined in rectangular microchannels. A plastic or synthetic wall separates the solution channels between them. The solution entering the absorber is previously subcooled using ambient air. In this way, the need for a cooling tower is avoided. A model of the configuration proposed is developed based on mass and energy balances and some correlations were selected to predict the heat and mass transfer coefficients. The concentration and temperatures along the channels cannot be explicitly determined from the set of equations obtained. For this reason, the equations were implemented in a computer code using Engineering Equation Solver software, EES™. With the aim of minimizing the absorber volume to reduce the size of absorption cooling chillers, the ratio between the cooling power of the chiller and the absorber volume (R) is calculated. Its variation is shown along the solution channels, allowing its optimization for selected operating conditions. For the case considered the solution channel length is recommended to be lower than 3 cm. Maximum values of R obtained in this work are higher than the ones found in optimized horizontal falling film absorbers using the same solution. Results obtained also show the variation of R and the chiller efficiency (COP) for different ambient temperatures and desorption temperatures typically obtained using flat plate solar collectors. The configuration proposed of adiabatic membrane-based absorber using ambient air to subcool the solution is a good technology to reduce the size of the absorption chillers, allowing the use of low temperature solar heat and avoiding the need for cooling towers.

Keywords: adiabatic absorption, air-cooled, membrane, solar thermal energy

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25 [Keynote Talk]: New Generations and Employment: An Exploratory Study about Tensions between the Psycho-Social Characteristics of the Generation Z and Expectations and Actions of Organizational Structures Related with Employment (CABA, 2016)

Authors: Esteban Maioli

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Generational studies have an important research tradition in social and human sciences. On the one hand, the speed of social change in the context of globalization imposes the need to research the transformations are identified both the subjectivity of the agents involved and its inclusion in the institutional matrix, specifically employment. Generation Z, (generally considered as the population group whose birth occurs after 1995) have unique psycho-social characteristics. Gen Z is characterized by a different set of values, beliefs, attitudes and ambitions that impact in their concrete action in organizational structures. On the other hand, managers often have to deal with generational differences in the workplace. Organizations have members who belong to different generations; they had never before faced the challenge of having such a diverse group of members. The members of each historical generation are characterized by a different set of values, beliefs, attitudes and ambitions that are manifest in their concrete action in organizational structures. Gen Z it’s the only one who can fully be considered "global," while its members were born in the consolidated context of globalization. Some salient features of the Generation Z can be summarized as follows. They’re the first fully born into a digital world. Social networks and technology are integrated into their lives. They are concerned about the challenges of the modern world (poverty, inequality, climate change, among others). They are self-expressive, more liberal and open to change. They often bore easily, with short attention spans. They do not like routine tasks. They want to achieve a good life-work balance, and they are interested in a flexible work environment, as opposed to traditional work schedule. They are critical thinkers, who come with innovative and creative ideas to help. Research design considered methodological triangulation. Data was collected with two techniques: a self-administered survey with multiple choice questions and attitudinal scales applied over a non-probabilistic sample by reasoned decision. According to the multi-method strategy, also it was conducted in-depth interviews. Organizations constantly face new challenges. One of the biggest ones is to learn to manage a multi-generational scope of work. While Gen Z has not yet been fully incorporated (expected to do so in five years or so), many organizations have already begun to implement a series of changes in its recruitment and development. The main obstacle to retaining young talent is the gap between the expectations of iGen applicants and what companies offer. Members of the iGen expect not only a good salary and job stability but also a clear career plan. Generation Z needs to have immediate feedback on their tasks. However, many organizations have yet to improve both motivation and monitoring practices. It is essential for companies to take a review of organizational practices anchored in the culture of the organization.

Keywords: employment, expectations, generation Z, organizational culture, organizations, psycho-social characteristics

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24 Laparoscopic Resection Shows Comparable Outcomes to Open Thoracotomy for Thoracoabdominal Neuroblastomas: A Meta-Analysis and Systematic Review

Authors: Peter J. Fusco, Dave M. Mathew, Chris Mathew, Kenneth H. Levy, Kathryn S. Varghese, Stephanie Salazar-Restrepo, Serena M. Mathew, Sofia Khaja, Eamon Vega, Mia Polizzi, Alyssa Mullane, Adham Ahmed

Abstract:

Background: Laparoscopic (LS) removal of neuroblastomas in children has been reported to offer favorable outcomes compared to the conventional open thoracotomy (OT) procedure. Critical perioperative measures such as blood loss, operative time, length of stay, and time to postoperative chemotherapy have all supported laparoscopic use rather than its more invasive counterpart. Herein, a pairwise meta-analysis was performed comparing perioperative outcomes between LS and OT in thoracoabdominal neuroblastoma cases. Methods: A comprehensive literature search was performed on PubMed, Ovid EMBASE, and Scopus databases to identify studies comparing the outcomes of pediatric patients with thoracoabdominal neuroblastomas undergoing resection via OT or LS. After deduplication, 4,227 studies were identified and subjected to initial title screening with exclusion and inclusion criteria to ensure relevance. When studies contained overlapping cohorts, only the larger series were included. Primary outcomes include estimated blood loss (EBL), hospital length of stay (LOS), and mortality, while secondary outcomes were tumor recurrence, post-operative complications, and operation length. The “meta” and “metafor” packages were used in R, version 4.0.2, to pool risk ratios (RR) or standardized mean differences (SMD) in addition to their 95% confidence intervals in the random effects model via the Mantel-Haenszel method. Heterogeneity between studies was assessed using the I² test, while publication bias was assessed via funnel plot. Results: The pooled analysis included 209 patients from 5 studies (141 OT, 68 LS). Of the included studies, 2 originated from the United States, 1 from Toronto, 1 from China, and 1was from a Japanese center. Mean age between study cohorts ranged from 2.4 to 5.3 years old, with female patients occupying between 30.8% to 50% of the study populations. No statistically significant difference was found between the two groups for LOS (SMD -1.02; p=0.083), mortality (RR 0.30; p=0.251), recurrence(RR 0.31; p=0.162), post-operative complications (RR 0.73; p=0.732), or operation length (SMD -0.07; p=0.648). Of note, LS appeared to be protective in the analysis for EBL, although it did not reach statistical significance (SMD -0.4174; p= 0.051). Conclusion: Despite promising literature assessing LS removal of pediatric neuroblastomas, results showed it was non-superior to OT for any explored perioperative outcomes. Given the limited comparative data on the subject, it is evident that randomized trials are necessary to further the efficacy of the conclusions reached.

Keywords: laparoscopy, neuroblastoma, thoracoabdominal, thoracotomy

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23 Development and Structural Characterization of a Snack Food with Added Type 4 Extruded Resistant Starch

Authors: Alberto A. Escobar Puentes, G. Adriana García, Luis F. Cuevas G., Alejandro P. Zepeda, Fernando B. Martínez, Susana A. Rincón

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Snack foods are usually classified as ‘junk food’ because have little nutritional value. However, due to the increase on the demand and third generation (3G) snacks market, low price and easy to prepare, can be considered as carriers of compounds with certain nutritional value. Resistant starch (RS) is classified as a prebiotic fiber it helps to control metabolic problems and has anti-cancer colon properties. The active compound can be developed by chemical cross-linking of starch with phosphate salts to obtain a type 4 resistant starch (RS4). The chemical reaction can be achieved by extrusion, a process widely used to produce snack foods, since it's versatile and a low-cost procedure. Starch is the major ingredient for snacks 3G manufacture, and the seeds of sorghum contain high levels of starch (70%), the most drought-tolerant gluten-free cereal. Due to this, the aim of this research was to develop a snack (3G), with RS4 in optimal conditions extrusion (previously determined) from sorghum starch, and carry on a sensory, chemically and structural characterization. A sample (200 g) of sorghum starch was conditioned with 4% sodium trimetaphosphate/ sodium tripolyphosphate (99:1) and set to 28.5% of moisture content. Then, the sample was processed in a single screw extruder equipped with rectangular die. The inlet, transport and output temperatures were 60°C, 134°C and 70°C, respectively. The resulting pellets were expanded in a microwave oven. The expansion index (EI), penetration force (PF) and sensory analysis were evaluated in the expanded pellets. The pellets were milled to obtain flour and RS content, degree of substitution (DS), and percentage of phosphorus (% P) were measured. Spectroscopy [Fourier Transform Infrared (FTIR)], X-ray diffraction, differential scanning calorimetry (DSC) and scanning electron microscopy (SEM) analysis were performed in order to determine structural changes after the process. The results in 3G were as follows: RS, 17.14 ± 0.29%; EI, 5.66 ± 0.35 and PF, 5.73 ± 0.15 (N). Groups of phosphate were identified in the starch molecule by FTIR: DS, 0.024 ± 0.003 and %P, 0.35±0.15 [values permitted as food additives (<4 %P)]. In this work an increase of the gelatinization temperature after the crosslinking of starch was detected; the loss of granular and vapor bubbles after expansion were observed by SEM; By using X-ray diffraction, loss of crystallinity was observed after extrusion process. Finally, a snack (3G) was obtained with RS4 developed by extrusion technology. The sorghum starch was efficient for snack 3G production.

Keywords: extrusion, resistant starch, snack (3G), Sorghum

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22 Resilience-Vulnerability Interaction in the Context of Disasters and Complexity: Study Case in the Coastal Plain of Gulf of Mexico

Authors: Cesar Vazquez-Gonzalez, Sophie Avila-Foucat, Leonardo Ortiz-Lozano, Patricia Moreno-Casasola, Alejandro Granados-Barba

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In the last twenty years, academic and scientific literature has been focused on understanding the processes and factors of coastal social-ecological systems vulnerability and resilience. Some scholars argue that resilience and vulnerability are isolated concepts due to their epistemological origin, while others note the existence of a strong resilience-vulnerability relationship. Here we present an ordinal logistic regression model based on the analytical framework about dynamic resilience-vulnerability interaction along adaptive cycle of complex systems and disasters process phases (during, recovery and learning). In this way, we demonstrate that 1) during the disturbance, absorptive capacity (resilience as a core of attributes) and external response capacity explain the probability of households capitals to diminish the damage, and exposure sets the thresholds about the amount of disturbance that households can absorb, 2) at recovery, absorptive capacity and external response capacity explain the probability of households capitals to recovery faster (resilience as an outcome) from damage, and 3) at learning, adaptive capacity (resilience as a core of attributes) explains the probability of households adaptation measures based on the enhancement of physical capital. As a result, during the disturbance phase, exposure has the greatest weight in the probability of capital’s damage, and households with absorptive and external response capacity elements absorbed the impact of floods in comparison with households without these elements. At the recovery phase, households with absorptive and external response capacity showed a faster recovery on their capital; however, the damage sets the thresholds of recovery time. More importantly, diversity in financial capital increases the probability of recovering other capital, but it becomes a liability so that the probability of recovering the household finances in a longer time increases. At learning-reorganizing phase, adaptation (modifications to the house) increases the probability of having less damage on physical capital; however, it is not very relevant. As conclusion, resilience is an outcome but also core of attributes that interacts with vulnerability along the adaptive cycle and disaster process phases. Absorptive capacity can diminish the damage experienced by floods; however, when exposure overcomes thresholds, both absorptive and external response capacity are not enough. In the same way, absorptive and external response capacity diminish the recovery time of capital, but the damage sets the thresholds in where households are not capable of recovering their capital.

Keywords: absorptive capacity, adaptive capacity, capital, floods, recovery-learning, social-ecological systems

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21 Improving Patient Outcomes for Aspiration Pneumonia

Authors: Mary Farrell, Maria Soubra, Sandra Vega, Dorothy Kakraba, Joanne Fontanilla, Moira Kendra, Danielle Tonzola, Stephanie Chiu

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Pneumonia is the most common infectious cause of hospitalizations in the United States, with more than one million admissions annually and costs of $10 billion every year, making it the 8th leading cause of death. Aspiration pneumonia is an aggressive type of pneumonia that results from inhalation of oropharyngeal secretions and/or gastric contents and is preventable. The authors hypothesized that an evidence-based aspiration pneumonia clinical care pathway could reduce 30-day hospital readmissions and mortality rates, while improving the overall care of patients. We conducted a retrospective chart review on 979 patients discharged with aspiration pneumonia from January 2021 to December 2022 at Overlook Medical Center. The authors identified patients who were coded with aspiration pneumonia and/or stable sepsis. Secondarily, we identified 30-day readmission rates for aspiration pneumonia from a SNF. The Aspiration Pneumonia Clinical Care Pathway starts in the emergency department (ED) with the initiation of antimicrobials within 4 hours of admission and early recognition of aspiration. Once this is identified, a swallow test is initiated by the bedside nurse, and if the patient demonstrates dysphagia, they are maintained on strict nothing by mouth (NPO) followed by a speech and language pathologist (SLP) referral for an appropriate modified diet recommendation. Aspiration prevention techniques included the avoidance of straws, 45-degree positioning, no talking during meals, taking small bites, placement of the aspiration wrist band, and consuming meals out of the bed in a chair. Nursing education was conducted with a newly created online learning module about aspiration pneumonia. The authors identified 979 patients, with an average age of 73.5 years old, who were diagnosed with aspiration pneumonia on the index hospitalization. These patients were reviewed for a 30-day readmission for aspiration pneumonia or stable sepsis, and mortality rates from January 2021 to December 2022 at Overlook Medical Center (OMC). The 30-day readmission rates were significantly lower in the cohort that received the clinical care pathway (35.0% vs. 27.5%, p = 0.011). When evaluating the mortality rates in the pre and post intervention cohort the authors discovered the mortality rates were lower in the post intervention cohort (23.7% vs 22.4%, p = 0.61) Mortality among non-white (self-reported as non-white) patients were lower in the post intervention cohort (34.4% vs. 21.0% , p = 0.05). Patients who reported as a current smoker/vaper in the pre and post cohorts had increased mortality rates (5.9% vs 22%). There was a decrease in mortality for the male population but an increase in mortality for women in the pre and post cohorts (19% vs. 25%). The authors attributed this increase in mortality in the post intervention cohort to more active smokers, more former smokers, and more being admitted from a SNF. This research identified that implementation of an Aspiration Pneumonia Clinical Care Pathway showed a statistically significant decrease in readmission rates and mortality rates in non-whites. The 30-day readmission rates were lower in the cohort that received the clinical care pathway (35.0% vs. 27.5%, p = 0.011).

Keywords: aspiration pneumonia, mortality, quality improvement, 30-day pneumonia readmissions

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20 Dendroremediation of a Defunct Lead Acid Battery Recycling Site

Authors: Alejandro Ruiz-Olivares, M. del Carmen González-Chávez, Rogelio Carrillo-González, Martha Reyes-Ramos, Javier Suárez Espinosa

Abstract:

Use of automobiles has increased and proportionally, the demand for batteries to impulse them. When the device is aged, all the battery materials are reused through lead acid battery recycling (LABR). Importation of used lead acid batteries in Mexico has increased in the last years since many recycling factories have been settled in the country. Inadequate disposal of lead-acid battery recycling (LABR) wastes left soil severely polluted with Pb, Cu, and salts (Na+, SO2− 4, PO3− 4). Soil organic amendments may contribute with essential nutrients and sequester (scavenger compounds) metals to allow plant establishment. The objective of this research was to revegetate a former lead-acid battery recycling site aided with organic amendments. Seven tree species (Acacia farnesiana, Casuarina equisetifolia, Cupressus lusitanica, Eucalyptus obliqua, Fraxinus excelsior, Prosopis laevigata and Pinus greggii) and two organic amendments (vermicompost and vermicompost + sawdust mixture) were tested for phytoremediation of a defunct LABR site. Plants were irrigated during the dry season. Monitoring of the soils was carried out during the experiment: Available metals, salts concentrations and their spatial pattern in soil were analyzed. Plant species and amendments were compared through analysis of covariance and longitudinal analysis. High concentrations of extractable (DTPA-TEA-CaCl₂) metals (up to 15,685 mg kg⁻¹ and 478 mg kg⁻¹ for Pb and Cu) and soluble salts (292 mg kg-1 and 23,578 mg kg-1 for PO3− 4and SO2− 4) were found in the soil after three and six months of setting up the experiment. Lead and Cu concentrations were depleted in the rhizosphere after amendments addition. Spatial pattern of PO3− 4, SO2− 4 and DTPA-extractable Pb and Cu changed slightly through time. In spite of extreme soil conditions the plant species planted: A. farnesiana, E. obliqua, C. equisetifolia and F. excelsior had 100% of survival. Available metals and salts differently affected each species. In addition, negative effect on growth due to Pb accumulated in shoots was observed only in C. lusitanica. Many specimens accumulated high concentrations of Pb ( > 1000 mg kg-1) in shoots. C. equisetifolia and C. lusitanica had the best rate of growth. Based on the results, all the evaluated species may be useful for revegetation of Pb-polluted soils. Besides their use in phytoremediation, some ecosystem services can be obtained from the woodland such as encourage wildlife, wood production, and carbon sequestration. Further research should be conducted to analyze these services.

Keywords: heavy metals, inadequate disposal, organic amendments, phytoremediation with trees

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19 Kinetic Evaluation of Sterically Hindered Amines under Partial Oxy-Combustion Conditions

Authors: Sara Camino, Fernando Vega, Mercedes Cano, Benito Navarrete, José A. Camino

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Carbon capture and storage (CCS) technologies should play a relevant role towards low-carbon systems in the European Union by 2030. Partial oxy-combustion emerges as a promising CCS approach to mitigate anthropogenic CO₂ emissions. Its advantages respect to other CCS technologies rely on the production of a higher CO₂ concentrated flue gas than these provided by conventional air-firing processes. The presence of more CO₂ in the flue gas increases the driving force in the separation process and hence it might lead to further reductions of the energy requirements of the overall CO₂ capture process. A higher CO₂ concentrated flue gas should enhance the CO₂ capture by chemical absorption in solvent kinetic and CO₂ cyclic capacity. They have impact on the performance of the overall CO₂ absorption process by reducing the solvent flow-rate required for a specific CO₂ removal efficiency. Lower solvent flow-rates decreases the reboiler duty during the regeneration stage and also reduces the equipment size and pumping costs. Moreover, R&D activities in this field are focused on novel solvents and blends that provide lower CO₂ absorption enthalpies and therefore lower energy penalties associated to the solvent regeneration. In this respect, sterically hindered amines are considered potential solvents for CO₂ capture. They provide a low energy requirement during the regeneration process due to its molecular structure. However, its absorption kinetics are slow and they must be promoted by blending with faster solvents such as monoethanolamine (MEA) and piperazine (PZ). In this work, the kinetic behavior of two sterically hindered amines were studied under partial oxy-combustion conditions and compared with MEA. A lab-scale semi-batch reactor was used. The CO₂ composition of the synthetic flue gas varied from 15%v/v – conventional coal combustion – to 60%v/v – maximum CO₂ concentration allowable for an optimal partial oxy-combustion operation. Firstly, 2-amino-2-methyl-1-propanol (AMP) showed a hybrid behavior with fast kinetics and a low enthalpy of CO₂ absorption. The second solvent was Isophrondiamine (IF), which has a steric hindrance in one of the amino groups. Its free amino group increases its cyclic capacity. In general, the presence of higher CO₂ concentration in the flue gas accelerated the CO₂ absorption phenomena, producing higher CO₂ absorption rates. In addition, the evolution of the CO2 loading also exhibited higher values in the experiments using higher CO₂ concentrated flue gas. The steric hindrance causes a hybrid behavior in this solvent, between both fast and slow kinetic solvents. The kinetics rates observed in all the experiments carried out using AMP were higher than MEA, but lower than the IF. The kinetic enhancement experienced by AMP at a high CO2 concentration is slightly over 60%, instead of 70% – 80% for IF. AMP also improved its CO₂ absorption capacity by 24.7%, from 15%v/v to 60%v/v, almost double the improvements achieved by MEA. In IF experiments, the CO₂ loading increased around 10% from 15%v/v to 60%v/v CO₂ and it changed from 1.10 to 1.34 mole CO₂ per mole solvent, more than 20% of increase. This hybrid kinetic behavior makes AMP and IF promising solvents for partial oxy–combustion applications.

Keywords: absorption, carbon capture, partial oxy-combustion, solvent

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18 Improving Student Learning in a Math Bridge Course through Computer Algebra Systems

Authors: Alejandro Adorjan

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Universities are motivated to understand the factor contributing to low retention of engineering undergraduates. While precollege students for engineering increases, the number of engineering graduates continues to decrease and attrition rates for engineering undergraduates remains high. Calculus 1 (C1) is the entry point of most undergraduate Engineering Science and often a prerequisite for Computing Curricula courses. Mathematics continues to be a major hurdle for engineering students and many students who drop out from engineering cite specifically Calculus as one of the most influential factors in that decision. In this context, creating course activities that increase retention and motivate students to obtain better final results is a challenge. In order to develop several competencies in our students of Software Engineering courses, Calculus 1 at Universidad ORT Uruguay focuses on developing several competencies such as capacity of synthesis, abstraction, and problem solving (based on the ACM/AIS/IEEE). Every semester we try to reflect on our practice and try to answer the following research question: What kind of teaching approach in Calculus 1 can we design to retain students and obtain better results? Since 2010, Universidad ORT Uruguay offers a six-week summer noncompulsory bridge course of preparatory math (to bridge the math gap between high school and university). Last semester was the first time the Department of Mathematics offered the course while students were enrolled in C1. Traditional lectures in this bridge course lead to just transcribe notes from blackboard. Last semester we proposed a Hands On Lab course using Geogebra (interactive geometry and Computer Algebra System (CAS) software) as a Math Driven Development Tool. Students worked in a computer laboratory class and developed most of the tasks and topics in Geogebra. As a result of this approach, several pros and cons were found. It was an excessive amount of weekly hours of mathematics for students and, as the course was non-compulsory; the attendance decreased with time. Nevertheless, this activity succeeds in improving final test results and most students expressed the pleasure of working with this methodology. This teaching technology oriented approach strengthens student math competencies needed for Calculus 1 and improves student performance, engagement, and self-confidence. It is important as a teacher to reflect on our practice, including innovative proposals with the objective of engaging students, increasing retention and obtaining better results. The high degree of motivation and engagement of participants with this methodology exceeded our initial expectations, so we plan to experiment with more groups during the summer so as to validate preliminary results.

Keywords: calculus, engineering education, PreCalculus, Summer Program

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17 Sorghum Polyphenols Encapsulated by Spray Drying, Using Modified Starches as Wall Materials

Authors: Adriana Garcia G., Alberto A. Escobar P., Amira D. Calvo L., Gabriel Lizama U., Alejandro Zepeda P., Fernando Martínez B., Susana Rincón A.

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Different studies have recently been focused on the use of antioxidants such as polyphenols because of to its anticarcinogenic capacity. However, these compounds are highly sensible to environmental factors such as light and heat, so lose its long-term stability, besides possess an astringent and bitter taste. Nevertheless, the polyphenols can be protected by microcapsule formulation. In this sense, a rich source of polyphenols is sorghum, besides presenting a high starch content. Due to the above, the aim of this work was to obtain modified starches from sorghum by extrusion to encapsulate polyphenols the sorghum by spray drying. Polyphenols were extracted by ethanol solution from sorghum (Pajarero/red) and determined by the method of Folin-Ciocalteu, obtaining GAE at 30 mg/g. Moreover, was extracted starch of sorghum (Sinaloense/white) through wet milling (yield 32 %). The hydrolyzed starch was modified with three treatments: acetic anhydride (2.5g/100g), sodium tripolyphosphate (4g/100g), and sodium tripolyphosphate/ acetic anhydride (2g/1.25g by each 100 g) by extrusion. Processing conditions of extrusion were as follows: barrel temperatures were of 60, 130 and 170 °C at the feeding, transition, and high-pressure extrusion zones, respectively. Analysis of Fourier Transform Infrared spectroscopy (FTIR), showed bands exhibited of acetyl groups (1735 cm-1) and phosphates (1170 cm-1, 910 cm-1 and 525 cm-1), indicating the respective modification of starch. Besides, all modified starches not developed viscosity, which is a characteristic required for use in the encapsulation of polyphenols using the spray drying technique. As result of the modification starch, was obtained a water solubility index (WSI) from 33.8 to 44.8 %, and crystallinity from 8 to 11 %, indicating the destruction of the starch granule. Afterwards, microencapsulation of polyphenols was developed by spray drying, with a blend of 10 g of modified starch, 60 ml polyphenol extract and 30 ml of distilled water. Drying conditions were as follows: inlet air temperature 150 °C ± 1, outlet air temperature 80°C ± 5. As result of the microencapsulation: were obtained yields of 56.8 to 77.4 % and an efficiency of encapsulation from 84.6 to 91.4 %. The FTIR analysis showed evidence of microcapsules loaded with polyphenols in bands 1042 cm-1, 1038 cm-1 and 1148 cm-1. Analysis Differential scanning calorimetry (DSC) showed transition temperatures from 144.1 to 173.9 °C. For the order hand, analysis of Scanning Electron Microscopy (SEM), were observed rounded surfaces with concavities, typical feature of microcapsules produced by spray drying, how result of rapid evaporation of water. Finally, the modified starches were obtained by extrusion with good characteristics for use as cover materials by spray drying, where the phosphorylated starch was the best treatment in this work, according to the encapsulation yield, efficiency, and transition temperature.

Keywords: encapsulation, extrusion, modified starch, polyphenols, spray drying

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16 A 500 MWₑ Coal-Fired Power Plant Operated under Partial Oxy-Combustion: Methodology and Economic Evaluation

Authors: Fernando Vega, Esmeralda Portillo, Sara Camino, Benito Navarrete, Elena Montavez

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The European Union aims at strongly reducing their CO₂ emissions from energy and industrial sector by 2030. The energy sector contributes with more than two-thirds of the CO₂ emission share derived from anthropogenic activities. Although efforts are mainly focused on the use of renewables by energy production sector, carbon capture and storage (CCS) remains as a frontline option to reduce CO₂ emissions from industrial process, particularly from fossil-fuel power plants and cement production. Among the most feasible and near-to-market CCS technologies, namely post-combustion and oxy-combustion, partial oxy-combustion is a novel concept that can potentially reduce the overall energy requirements of the CO₂ capture process. This technology consists in the use of higher oxygen content in the oxidizer that should increase the CO₂ concentration of the flue gas once the fuel is burnt. The CO₂ is then separated from the flue gas downstream by means of a conventional CO₂ chemical absorption process. The production of a higher CO₂ concentrated flue gas should enhance the CO₂ absorption into the solvent, leading to further reductions of the CO₂ separation performance in terms of solvent flow-rate, equipment size, and energy penalty related to the solvent regeneration. This work evaluates a portfolio of CCS technologies applied to fossil-fuel power plants. For this purpose, an economic evaluation methodology was developed in detail to determine the main economical parameters for CO₂ emission removal such as the levelized cost of electricity (LCOE) and the CO₂ captured and avoided costs. ASPEN Plus™ software was used to simulate the main units of power plant and solve the energy and mass balance. Capital and investment costs were determined from the purchased cost of equipment, also engineering costs and project and process contingencies. The annual capital cost and operating and maintenance costs were later obtained. A complete energy balance was performed to determine the net power produced in each case. The baseline case consists of a supercritical 500 MWe coal-fired power plant using anthracite as a fuel without any CO₂ capture system. Four cases were proposed: conventional post-combustion capture, oxy-combustion and partial oxy-combustion using two levels of oxygen-enriched air (40%v/v and 75%v/v). CO₂ chemical absorption process using monoethanolamine (MEA) was used as a CO₂ separation process whereas the O₂ requirement was achieved using a conventional air separation unit (ASU) based on Linde's cryogenic process. Results showed a reduction of 15% of the total investment cost of the CO₂ separation process when partial oxy-combustion was used. Oxygen-enriched air production also reduced almost half the investment costs required for ASU in comparison with oxy-combustion cases. Partial oxy-combustion has a significant impact on the performance of both CO₂ separation and O₂ production technologies, and it can lead to further energy reductions using new developments on both CO₂ and O₂ separation processes.

Keywords: carbon capture, cost methodology, economic evaluation, partial oxy-combustion

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15 Adding a Degree of Freedom to Opinion Dynamics Models

Authors: Dino Carpentras, Alejandro Dinkelberg, Michael Quayle

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Within agent-based modeling, opinion dynamics is the field that focuses on modeling people's opinions. In this prolific field, most of the literature is dedicated to the exploration of the two 'degrees of freedom' and how they impact the model’s properties (e.g., the average final opinion, the number of final clusters, etc.). These degrees of freedom are (1) the interaction rule, which determines how agents update their own opinion, and (2) the network topology, which defines the possible interaction among agents. In this work, we show that the third degree of freedom exists. This can be used to change a model's output up to 100% of its initial value or to transform two models (both from the literature) into each other. Since opinion dynamics models are representations of the real world, it is fundamental to understand how people’s opinions can be measured. Even for abstract models (i.e., not intended for the fitting of real-world data), it is important to understand if the way of numerically representing opinions is unique; and, if this is not the case, how the model dynamics would change by using different representations. The process of measuring opinions is non-trivial as it requires transforming real-world opinion (e.g., supporting most of the liberal ideals) to a number. Such a process is usually not discussed in opinion dynamics literature, but it has been intensively studied in a subfield of psychology called psychometrics. In psychometrics, opinion scales can be converted into each other, similarly to how meters can be converted to feet. Indeed, psychometrics routinely uses both linear and non-linear transformations of opinion scales. Here, we analyze how this transformation affects opinion dynamics models. We analyze this effect by using mathematical modeling and then validating our analysis with agent-based simulations. Firstly, we study the case of perfect scales. In this way, we show that scale transformations affect the model’s dynamics up to a qualitative level. This means that if two researchers use the same opinion dynamics model and even the same dataset, they could make totally different predictions just because they followed different renormalization processes. A similar situation appears if two different scales are used to measure opinions even on the same population. This effect may be as strong as providing an uncertainty of 100% on the simulation’s output (i.e., all results are possible). Still, by using perfect scales, we show that scales transformations can be used to perfectly transform one model to another. We test this using two models from the standard literature. Finally, we test the effect of scale transformation in the case of finite precision using a 7-points Likert scale. In this way, we show how a relatively small-scale transformation introduces both changes at the qualitative level (i.e., the most shared opinion at the end of the simulation) and in the number of opinion clusters. Thus, scale transformation appears to be a third degree of freedom of opinion dynamics models. This result deeply impacts both theoretical research on models' properties and on the application of models on real-world data.

Keywords: degrees of freedom, empirical validation, opinion scale, opinion dynamics

Procedia PDF Downloads 99
14 Validation and Fit of a Biomechanical Bipedal Walking Model for Simulation of Loads Induced by Pedestrians on Footbridges

Authors: Dianelys Vega, Carlos Magluta, Ney Roitman

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The simulation of loads induced by walking people in civil engineering structures is still challenging It has been the focus of considerable research worldwide in the recent decades due to increasing number of reported vibration problems in pedestrian structures. One of the most important key in the designing of slender structures is the Human-Structure Interaction (HSI). How moving people interact with structures and the effect it has on their dynamic responses is still not well understood. To rely on calibrated pedestrian models that accurately estimate the structural response becomes extremely important. However, because of the complexity of the pedestrian mechanisms, there are still some gaps in knowledge and more reliable models need to be investigated. On this topic several authors have proposed biodynamic models to represent the pedestrian, whether these models provide a consistent approximation to physical reality still needs to be studied. Therefore, this work comes to contribute to a better understanding of this phenomenon bringing an experimental validation of a pedestrian walking model and a Human-Structure Interaction model. In this study, a bi-dimensional bipedal walking model was used to represent the pedestrians along with an interaction model which was applied to a prototype footbridge. Numerical models were implemented in MATLAB. In parallel, experimental tests were conducted in the Structures Laboratory of COPPE (LabEst), at Federal University of Rio de Janeiro. Different test subjects were asked to walk at different walking speeds over instrumented force platforms to measure the walking force and an accelerometer was placed at the waist of each subject to measure the acceleration of the center of mass at the same time. By fitting the step force and the center of mass acceleration through successive numerical simulations, the model parameters are estimated. In addition, experimental data of a walking pedestrian on a flexible structure was used to validate the interaction model presented, through the comparison of the measured and simulated structural response at mid span. It was found that the pedestrian model was able to adequately reproduce the ground reaction force and the center of mass acceleration for normal and slow walking speeds, being less efficient for faster speeds. Numerical simulations showed that biomechanical parameters such as leg stiffness and damping affect the ground reaction force, and the higher the walking speed the greater the leg length of the model. Besides, the interaction model was also capable to estimate with good approximation the structural response, that remained in the same order of magnitude as the measured response. Some differences in frequency spectra were observed, which are presumed to be due to the perfectly periodic loading representation, neglecting intra-subject variabilities. In conclusion, this work showed that the bipedal walking model could be used to represent walking pedestrians since it was efficient to reproduce the center of mass movement and ground reaction forces produced by humans. Furthermore, although more experimental validations are required, the interaction model also seems to be a useful framework to estimate the dynamic response of structures under loads induced by walking pedestrians.

Keywords: biodynamic models, bipedal walking models, human induced loads, human structure interaction

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13 Displaying Compostela: Literature, Tourism and Cultural Representation, a Cartographic Approach

Authors: Fernando Cabo Aseguinolaza, Víctor Bouzas Blanco, Alberto Martí Ezpeleta

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Santiago de Compostela became a stable object of literary representation during the period between 1840 and 1915, approximately. This study offers a partial cartographical look at this process, suggesting that a cultural space like Compostela’s becoming an object of literary representation paralleled the first stages of its becoming a tourist destination. We use maps as a method of analysis to show the interaction between a corpus of novels and the emerging tradition of tourist guides on Compostela during the selected period. Often, the novels constitute ways to present a city to the outside, marking it for the gaze of others, as guidebooks do. That leads us to examine the ways of constructing and rendering communicable the local in other contexts. For that matter, we should also acknowledge the fact that a good number of the narratives in the corpus evoke the representation of the city through the figure of one who comes from elsewhere: a traveler, a student or a professor. The guidebooks coincide in this with the emerging fiction, of which the mimesis of a city is a key characteristic. The local cannot define itself except through a process of symbolic negotiation, in which recognition and self-recognition play important roles. Cartography shows some of the forms that these processes of symbolic representation take through the treatment of space. The research uses GIS to find significant models of representation. We used the program ArcGIS for the mapping, defining the databases starting from an adapted version of the methodology applied by Barbara Piatti and Lorenz Hurni’s team at the University of Zurich. First, we designed maps that emphasize the peripheral position of Compostela from a historical and institutional perspective using elements found in the texts of our corpus (novels and tourist guides). Second, other maps delve into the parallels between recurring techniques in the fictional texts and characteristic devices of the guidebooks (sketching itineraries and the selection of zones and indexicalization), like a foreigner’s visit guided by someone who knows the city or the description of one’s first entrance into the city’s premises. Last, we offer a cartography that demonstrates the connection between the best known of the novels in our corpus (Alejandro Pérez Lugín’s 1915 novel La casa de la Troya) and the first attempt to create package tourist tours with Galicia as a destination, in a joint venture of Galician and British business owners, in the years immediately preceding the Great War. Literary cartography becomes a crucial instrument for digging deeply into the methods of cultural production of places. Through maps, the interaction between discursive forms seemingly so far removed from each other as novels and tourist guides becomes obvious and suggests the need to go deeper into a complex process through which a city like Compostela becomes visible on the contemporary cultural horizon.

Keywords: compostela, literary geography, literary cartography, tourism

Procedia PDF Downloads 365
12 Methodology to Achieve Non-Cooperative Target Identification Using High Resolution Range Profiles

Authors: Olga Hernán-Vega, Patricia López-Rodríguez, David Escot-Bocanegra, Raúl Fernández-Recio, Ignacio Bravo

Abstract:

Non-Cooperative Target Identification has become a key research domain in the Defense industry since it provides the ability to recognize targets at long distance and under any weather condition. High Resolution Range Profiles, one-dimensional radar images where the reflectivity of a target is projected onto the radar line of sight, are widely used for identification of flying targets. According to that, to face this problem, an approach to Non-Cooperative Target Identification based on the exploitation of Singular Value Decomposition to a matrix of range profiles is presented. Target Identification based on one-dimensional radar images compares a collection of profiles of a given target, namely test set, with the profiles included in a pre-loaded database, namely training set. The classification is improved by using Singular Value Decomposition since it allows to model each aircraft as a subspace and to accomplish recognition in a transformed domain where the main features are easier to extract hence, reducing unwanted information such as noise. Singular Value Decomposition permits to define a signal subspace which contain the highest percentage of the energy, and a noise subspace which will be discarded. This way, only the valuable information of each target is used in the recognition process. The identification algorithm is based on finding the target that minimizes the angle between subspaces and takes place in a transformed domain. Two metrics, F1 and F2, based on Singular Value Decomposition are accomplished in the identification process. In the case of F2, the angle is weighted, since the top vectors set the importance in the contribution to the formation of a target signal, on the contrary F1 simply shows the evolution of the unweighted angle. In order to have a wide database or radar signatures and evaluate the performance, range profiles are obtained through numerical simulation of seven civil aircraft at defined trajectories taken from an actual measurement. Taking into account the nature of the datasets, the main drawback of using simulated profiles instead of actual measured profiles is that the former implies an ideal identification scenario, since measured profiles suffer from noise, clutter and other unwanted information and simulated profiles don't. In this case, the test and training samples have similar nature and usually a similar high signal-to-noise ratio, so as to assess the feasibility of the approach, the addition of noise has been considered before the creation of the test set. The identification results applying the unweighted and weighted metrics are analysed for demonstrating which algorithm provides the best robustness against noise in an actual possible scenario. So as to confirm the validity of the methodology, identification experiments of profiles coming from electromagnetic simulations are conducted, revealing promising results. Considering the dissimilarities between the test and training sets when noise is added, the recognition performance has been improved when weighting is applied. Future experiments with larger sets are expected to be conducted with the aim of finally using actual profiles as test sets in a real hostile situation.

Keywords: HRRP, NCTI, simulated/synthetic database, SVD

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11 Assessment of Surface Water Quality near Landfill Sites Using a Water Pollution Index

Authors: Alejandro Cittadino, David Allende

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Landfilling of municipal solid waste is a common waste management practice in Argentina as in many parts of the world. There is extensive scientific literature on the potential negative effects of landfill leachates on the environment, so it’s necessary to be rigorous with the control and monitoring systems. Due to the specific municipal solid waste composition in Argentina, local landfill leachates contain large amounts of organic matter (biodegradable, but also refractory to biodegradation), as well as ammonia-nitrogen, small trace of some heavy metals, and inorganic salts. In order to investigate the surface water quality in the Reconquista river adjacent to the Norte III landfill, water samples both upstream and downstream the dumpsite are quarterly collected and analyzed for 43 parameters including organic matter, heavy metals, and inorganic salts, as required by the local standards. The objective of this study is to apply a water quality index that considers the leachate characteristics in order to determine the quality status of the watercourse through the landfill. The water pollution index method has been widely used in water quality assessments, particularly rivers, and it has played an increasingly important role in water resource management, since it provides a number simple enough for the public to understand, that states the overall water quality at a certain location and time. The chosen water quality index (ICA) is based on the values of six parameters: dissolved oxygen (in mg/l and percent saturation), temperature, biochemical oxygen demand (BOD5), ammonia-nitrogen and chloride (Cl-) concentration. The index 'ICA' was determined both upstream and downstream the Reconquista river, being the rating scale between 0 (very poor water quality) and 10 (excellent water quality). The monitoring results indicated that the water quality was unaffected by possible leachate runoff since the index scores upstream and downstream were ranked in the same category, although in general, most of the samples were classified as having poor water quality according to the index’s scale. The annual averaged ICA index scores (computed quarterly) were 4.9, 3.9, 4.4 and 5.0 upstream and 3.9, 5.0, 5.1 and 5.0 downstream the river during the study period between 2014 and 2017. Additionally, the water quality seemed to exhibit distinct seasonal variations, probably due to annual precipitation patterns in the study area. The ICA water quality index appears to be appropriate to evaluate landfill impacts since it accounts mainly for organic pollution and inorganic salts and the absence of heavy metals in the local leachate composition, however, the inclusion of other parameters could be more decisive in discerning the affected stream reaches from the landfill activities. A future work may consider adding to the index other parameters like total organic carbon (TOC) and total suspended solids (TSS) since they are present in the leachate in high concentrations.

Keywords: landfill, leachate, surface water, water quality index

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10 Use of Zikani’s Ribosome Modulating Agents for Treating Recessive Dystrophic & Junctional Epidermolysis Bullosa with Nonsense Mutations

Authors: Mei Chen, Yingping Hou, Michelle Hao, Soheil Aghamohammadzadeh, Esteban Terzo, Roger Clark, Vijay Modur

Abstract:

Background: Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a genetic skin condition characterized by skin tearing and unremitting blistering upon minimal trauma. Repeated blistering, fibrosis, and scarring lead to aggressive squamous cell carcinoma later in life. RDEB is caused by mutations in the COL7A1 gene encoding collagen type VII (C7), the major component of anchoring fibrils mediating epidermis-dermis adherence. Nonsense mutations in the COL7A1 gene of a subset of RDEB patients leads to premature termination codons (PTC). Similarly, most Junctional Epidermolysis Bullosa (JEB) cases are caused by nonsense mutations in the LAMB3 gene encoding the β3 subunit of laminin 332. Currently, there is an unmet need for the treatment of RDEB and JEB. Zikani Therapeutics has discovered an array of macrocyclic compounds with ring structures similar to macrolide antibiotics that can facilitate readthrough activity of nonsense mutations in the COL7A1 and LAMB3 genes by acting as Ribosome Modulating Agents (RMAs). The medicinal chemistry synthetic advancements of these macrocyclic compounds have allowed targeting the human ribosome while preserving the structural elements responsible for the safety and pharmacokinetic profile of clinically used macrolide antibiotics. Methods: C7 expression was used as a measure of readthrough activity by immunoblot assays in two primary human fibroblasts from RDEB patients (R578X/R578X and R163X/R1683X-COL7A1). Similarly, immunoblot assays in C325X/c.629-12T > A-LAMB3 keratinocytes were used to measure readthrough activity for JEB. The relative readthrough activity of each compound was measured relative to Gentamicin. An imaging-based fibroblast migration assay was used as an assessment of C7 functionality in RDEB-fibroblasts over 16-20 hrs. The incubation period for the above experiments was 48 hrs for RDEB fibroblasts and 72 hours for JEB keratinocytes. Results: 9 RMAs demonstrated increased protein expression in both patient RDEB fibroblasts. The highest readthrough activity at tested concentrations without cytotoxicities increased protein expression up to 179% of Gentamicin (400 µg/ml), with favored readthrough activity in R163X/R1683X-COL7A1 fibroblasts. Concurrent with protein expression, fibroblast hypermotility phenotype observed in RDEB was rescued by reducing motility by ~35% to WT levels (the same level as 690 µM Gentamicin treated cells). Laminin β3 expression was also shown to be increased by 6 RMAs in keratinocytes to 33-83% of (400 µg/ml) Gentamicin. Conclusions: To date, 9 RMAs have been identified that enhance the expression of functional C7 in a mutation-dependent manner in two different RDEB patient fibroblast backgrounds (R578X/R578X and R163X/R1683X-COL7A1). A further 6 RMAs have been identified that enhance the readthrough of C325X-LAMB3 in JEB patient keratinocytes. Based on the clinical trial conducted by us with topical gentamycin in 2017, Zikani’s RMAs achieve clinically significant levels of read-through for the treatment of recessive dystrophic and Junctional Epidermolysis Bullosa.

Keywords: epidermolysis bullosa, nonsense mutation, readthrough, ribosome modulation

Procedia PDF Downloads 76
9 Argos-Linked Fastloc GPS Reveals the Resting Activity of Migrating Sea Turtles

Authors: Gail Schofield, Antoine M. Dujon, Nicole Esteban, Rebecca M. Lester, Graeme C. Hays

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Variation in diel movement patterns during migration provides information on the strategies used by animals to maximize energy efficiency and ensure the successful completion of migration. For instance, many flying and land-based terrestrial species stop to rest and refuel at regular intervals along the migratory route, or at transitory ‘stopover’ sites, depending on resource availability. However, in cases where stopping is not possible (such as over–or through deep–open oceans, or over deserts and mountains), non-stop travel is required, with animals needing to develop strategies to rest while actively traveling. Recent advances in biologging technologies have identified mid-flight micro sleeps by swifts in Africa during the 10-month non-breeding period, and the use of lateralized sleep behavior in orca and bottlenose dolphins during migration. Here, highly accurate locations obtained by Argos-linked Fastloc-GPS transmitters of adult green (n=8 turtles, 9487 locations) and loggerhead (n=46 turtles, 47,588 locations) sea turtles migrating around thousand kilometers (over several weeks) from breeding to foraging grounds across the Indian and Mediterranean oceans were used to identify potential resting strategies. Stopovers were only documented for seven turtles, lasting up to 6 days; thus, this strategy was not commonly used, possibly due to the lack of potential ‘shallow’ ( < 100 m seabed depth) sites along routes. However, observations of the day versus night speed of travel indicated that turtles might use other mechanisms to rest. For instance, turtles traveled an average 31% slower at night compared to day during oceanic crossings. Slower travel speeds at night might be explained by turtles swimming in a less direct line at night and/or deeper dives reducing their forward motion, as indicated through studies using Argos-linked transmitters and accelerometers. Furthermore, within the first 24 h of entering waters shallower than 100 m towards the end of migration (the depth at which sea turtles can swim and rest on the seabed), some individuals travelled 72% slower at night, repeating this behavior intermittently (each time for a one-night duration at 3–6-day intervals) until reaching the foraging grounds. If the turtles were, in fact, resting on the seabed at this point, they could be inactive for up to 8-hours, facilitating protracted periods of rest after several weeks of constant swimming. Turtles might not rest every night once within these shallower depths, due to the time constraints of reaching foraging grounds and restoring depleted energetic reserves (as sea turtles are capital breeders, they tend not to feed for several months during migration to and from the breeding grounds and while breeding). In conclusion, access to data-rich, highly accurate Argos-linked Fastloc-GPS provided information about differences in the day versus night activity at different stages of migration, allowing us, for the first time, to compare the strategies used by a marine vertebrate with terrestrial land-based and flying species. However, the question of what resting strategies are used by individuals that remain in oceanic waters to forage, with combinations of highly accurate Argos-linked Fastloc-GPS transmitters and accelerometry or time-depth recorders being required for sufficient numbers of individuals.

Keywords: argos-linked fastloc GPS, data loggers, migration, resting strategy, telemetry

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8 Nursery Treatments May Improve Restoration Outcomes by Reducing Seedling Transplant Shock

Authors: Douglas E. Mainhart, Alejandro Fierro-Cabo, Bradley Christoffersen, Charlotte Reemts

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Semi-arid ecosystems across the globe have faced land conversion for agriculture and resource extraction activities, posing a threat to the important ecosystem services they provide. Revegetation-centered restoration efforts in these regions face low success rates due to limited soil water availability and high temperatures leading to elevated seedling mortality after planting. Typical methods to alleviate these stresses require costly post-planting interventions aimed at improving soil moisture status. We set out to evaluate the efficacy of applying in-nursery treatments to address transplant shock. Four native Tamaulipan thornscrub species were compared. Three treatments were applied: elevated CO2, drought hardening (four-week exposure each), and antitranspirant foliar spray (the day prior to planting). Our goal was to answer two primary questions: (1) Do treatments improve survival and growth of seedlings in the early period post-planting? (2) If so, what underlying physiological changes are associated with this improved performance? To this end, we measured leaf gas exchange (stomatal conductance, light saturated photosynthetic rate, water use efficiency), leaf morphology (specific leaf area), and osmolality before and upon the conclusion of treatments. A subset of seedlings from all treatments have been planted, which will be monitored in coming months for in-field survival and growth.First month field survival for all treatment groups were high due to ample rainfall following planting (>85%). Growth data was unreliable due to high herbivory (68% of all sampled plants). While elevated CO2 had infrequent or no detectable influence on all aspects of leaf gas exchange, drought hardening reduced stomatal conductance in three of the four species measured without negatively impacting photosynthesis. Both CO2 and drought hardening elevated leaf osmolality in two species. Antitranspirant application significantly reduced conductance in all species for up to four days and reduced photosynthesis in two species. Antitranspirants also increased the variability of water use efficiency compared to controls. Collectively, these results suggest that antitranspirants and drought hardening are viable treatments for reducing short-term water loss during the transplant shock period. Elevated CO2, while not effective at reducing water loss, may be useful for promoting more favorable water status via osmotic adjustment. These practices could improve restoration outcomes in Tamaulipan thornscrub and other semi-arid systems. Further research should focus on evaluating combinations of these treatments and their species-specific viability.

Keywords: conservation, drought conditioning, semi-arid restoration, plant physiology

Procedia PDF Downloads 57
7 Numerical Analysis of the Computational Fluid Dynamics of Co-Digestion in a Large-Scale Continuous Stirred Tank Reactor

Authors: Sylvana A. Vega, Cesar E. Huilinir, Carlos J. Gonzalez

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Co-digestion in anaerobic biodigesters is a technology improving hydrolysis by increasing methane generation. In the present study, the dimensional computational fluid dynamics (CFD) is numerically analyzed using Ansys Fluent software for agitation in a full-scale Continuous Stirred Tank Reactor (CSTR) biodigester during the co-digestion process. For this, a rheological study of the substrate is carried out, establishing rotation speeds of the stirrers depending on the microbial activity and energy ranges. The substrate is organic waste from industrial sources of sanitary water, butcher, fishmonger, and dairy. Once the rheological behavior curves have been obtained, it is obtained that it is a non-Newtonian fluid of the pseudoplastic type, with a solids rate of 12%. In the simulation, the rheological results of the fluid are considered, and the full-scale CSTR biodigester is modeled. It was coupling the second-order continuity differential equations, the three-dimensional Navier Stokes, the power-law model for non-Newtonian fluids, and three turbulence models: k-ε RNG, k-ε Realizable, and RMS (Reynolds Stress Model), for a 45° tilt vane impeller. It is simulated for three minutes since it is desired to study an intermittent mixture with a saving benefit of energy consumed. The results show that the absolute errors of the power number associated with the k-ε RNG, k-ε Realizable, and RMS models were 7.62%, 1.85%, and 5.05%, respectively, the numbers of power obtained from the analytical-experimental equation of Nagata. The results of the generalized Reynolds number show that the fluid dynamics have a transition-turbulent flow regime. Concerning the Froude number, the result indicates there is no need to implement baffles in the biodigester design, and the power number provides a steady trend close to 1.5. It is observed that the levels of design speeds within the biodigester are approximately 0.1 m/s, which are speeds suitable for the microbial community, where they can coexist and feed on the substrate in co-digestion. It is concluded that the model that more accurately predicts the behavior of fluid dynamics within the reactor is the k-ε Realizable model. The flow paths obtained are consistent with what is stated in the referenced literature, where the 45° inclination PBT impeller is the right type of agitator to keep particles in suspension and, in turn, increase the dispersion of gas in the liquid phase. If a 24/7 complete mix is considered under stirred agitation, with a plant factor of 80%, 51,840 kWh/year are estimated. On the contrary, if intermittent agitations of 3 min every 15 min are used under the same design conditions, reduce almost 80% of energy costs. It is a feasible solution to predict the energy expenditure of an anaerobic biodigester CSTR. It is recommended to use high mixing intensities, at the beginning and end of the joint phase acetogenesis/methanogenesis. This high intensity of mixing, in the beginning, produces the activation of the bacteria, and once reaching the end of the Hydraulic Retention Time period, it produces another increase in the mixing agitations, favoring the final dispersion of the biogas that may be trapped in the biodigester bottom.

Keywords: anaerobic co-digestion, computational fluid dynamics, CFD, net power, organic waste

Procedia PDF Downloads 82
6 Absolute Quantification of the Bexsero Vaccine Component Factor H Binding Protein (fHbp) by Selected Reaction Monitoring: The Contribution of Mass Spectrometry in Vaccinology

Authors: Massimiliano Biagini, Marco Spinsanti, Gabriella De Angelis, Sara Tomei, Ilaria Ferlenghi, Maria Scarselli, Alessia Biolchi, Alessandro Muzzi, Brunella Brunelli, Silvana Savino, Marzia M. Giuliani, Isabel Delany, Paolo Costantino, Rino Rappuoli, Vega Masignani, Nathalie Norais

Abstract:

The gram-negative bacterium Neisseria meningitidis serogroup B (MenB) is an exclusively human pathogen representing the major cause of meningitides and severe sepsis in infants and children but also in young adults. This pathogen is usually present in the 30% of healthy population that act as a reservoir, spreading it through saliva and respiratory fluids during coughing, sneezing, kissing. Among surface-exposed protein components of this diplococcus, factor H binding protein is a lipoprotein proved to be a protective antigen used as a component of the recently licensed Bexsero vaccine. fHbp is a highly variable meningococcal protein: to reflect its remarkable sequence variability, it has been classified in three variants (or two subfamilies), and with poor cross-protection among the different variants. Furthermore, the level of fHbp expression varies significantly among strains, and this has also been considered an important factor for predicting MenB strain susceptibility to anti-fHbp antisera. Different methods have been used to assess fHbp expression on meningococcal strains, however, all these methods use anti-fHbp antibodies, and for this reason, the results are affected by the different affinity that antibodies can have to different antigenic variants. To overcome the limitations of an antibody-based quantification, we developed a quantitative Mass Spectrometry (MS) approach. Selected Reaction Monitoring (SRM) recently emerged as a powerful MS tool for detecting and quantifying proteins in complex mixtures. SRM is based on the targeted detection of ProteoTypicPeptides (PTPs), which are unique signatures of a protein that can be easily detected and quantified by MS. This approach, proven to be highly sensitive, quantitatively accurate and highly reproducible, was used to quantify the absolute amount of fHbp antigen in total extracts derived from 105 clinical isolates, evenly distributed among the three main variant groups and selected to be representative of the fHbp circulating subvariants around the world. We extended the study at the genetic level investigating the correlation between the differential level of expression and polymorphisms present within the genes and their promoter sequences. The implications of fHbp expression on the susceptibility of the strain to killing by anti-fHbp antisera are also presented. To date this is the first comprehensive fHbp expression profiling in a large panel of Neisseria meningitidis clinical isolates driven by an antibody-independent MS-based methodology, opening the door to new applications in vaccine coverage prediction and reinforcing the molecular understanding of released vaccines.

Keywords: quantitative mass spectrometry, Neisseria meningitidis, vaccines, bexsero, molecular epidemiology

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5 Nanocomposite Effect Based on Silver Nanoparticles and Anemposis Californica Extract as Skin Restorer

Authors: Maria Zulema Morquecho Vega, Fabiola CarolinaMiranda Castro, Rafael Verdugo Miranda, Ignacio Yocupicio Villegas, Ana lidia Barron Raygoza, Martin enrique MArquez Cordova, Jose Alberto Duarte Moller

Abstract:

Background: Anemopsis californica, also called (tame grass) belongs to the Saururaceae family small, green plant. The blade is long and wide. Gives a white flower. The plant population is only found in humid, swampy habitats, it grows where there is water, along the banks of streams and water holes. In the winter, it dries up. The leaves, rhizomes, or roots of this plant have been used to treat a range of diseases. Some of its healing properties are used to treat wounds, cold and flu symptoms, spasmodic cough, infection, pain and inflammation, burns, swollen feet, as well as lung ailments, asthma, circulatory problems (varicose veins), rheumatoid arthritis, purifies blood, helps in urinary and digestive tract diseases, sores and healing, for headache, sore throat, diarrhea, kidney pain. The tea made from the leaves and roots is used to treat uterine cancer, womb cancer, relieves menstrual pain and stops excessive bleeding after childbirth. It is also used as a gynecological treatment for infections, hemorrhoids, candidiasis and vaginitis. Objective: To study the cytotoxicity of gels prepared with silver nanoparticles in AC extract combined with chitosan, collagen and hyaluronic acid as an alternative therapy for skin conditions. Methods: The Ag NPs were synthesized according to the following method. A 0.3 mg/mL solution is prepared in 10 ml of deionized water, adjust to pH 12 with NaOH, stirring is maintained constant magnetic and a temperature of 80 °C. Subsequently, 100 ul of a 0.1 M AgNO3 solution and kept stirring constantly for 15 min. Once the reaction is complete, measurements are performed by UV-Vis. A gel was prepared in a 5% solution of acetic acid with the respective nanoparticles and AC extract of silver in the extract of AC. Chitosan is added until the process begins to occur gel. At that time, collagen will be added in a ratio of 3 to 5 drops, and later, hyaluronic acid in 2% of the total compound formed. Finally, after resting for 24 hours, the cytotoxic effect of the gels was studied. in the presence of highly positive bacteria Staphylococcus aureus and highly negative for Escherichia coli. Cultures will be incubated for 24 hours in the presence of the compound and compared with the reference. Results: Silver nanoparticles obtained had a spherical shape and sizes among 20 and 30 nm. UV-Vis spectra confirm the presence of silver nanoparticles showing a surface plasmon around 420 nm. Finally, the test in presence of bacteria yield a good antibacterial property of this nanocompound and tests in people were successful. Conclusion: Gel prepared by biogenic synthesis shown beneficious effects in severe acne, acne vulgaris and wound healing with diabetic patients.

Keywords: anemopsis californica, nanomedicina, biotechnology, biomedicine

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4 Delicate Balance between Cardiac Stress and Protection: Role of Mitochondrial Proteins

Authors: Zuzana Tatarkova, Ivana Pilchova, Michal Cibulka, Martin Kolisek, Peter Racay, Peter Kaplan

Abstract:

Introduction: Normal functioning of mitochondria is crucial for cardiac performance. Mitochondria undergo mitophagy and biogenesis, and mitochondrial proteins are subject to extensive post-translational modifications. The state of mitochondrial homeostasis reflects overall cellular fitness and longevity. Perturbed mitochondria produce less ATP, release greater amounts of reactive molecules, and are more prone to apoptosis. Therefore mitochondrial turnover is an integral aspect of quality control in which dysfunctional mitochondria are selectively eliminated through mitophagy. Currently, the progressive deterioration of physiological functions is seen as accumulation of modified/damaged proteins with limiting regenerative ability and disturbance of such affected protein-protein communication throughout aging in myocardial cells. Methodologies: For our study was used immunohistochemistry, biochemical methods: spectrophotometry, western blotting, immunodetection as well as more sophisticated 2D electrophoresis and mass spectrometry for evaluation protein-protein interactions and specific post-translational modification. Results and Discussion: Mitochondrial stress response to reactive species was evaluated as electron transport chain (ETC) complexes, redox-active molecules, and their possible communication. Protein-protein interactions revealed a strong linkage between age and ETC protein subunits. Redox state was strongly affected in senescent mitochondria with shift in favor of more pro-oxidizing condition within cardiomyocytes. Acute myocardial ischemia and ischemia-reperfusion (IR) injury affected ETC complexes I, II and IV with no change in complex III. Ischemia induced decrease in total antioxidant capacity, MnSOD, GSH and catalase activity with recovery in some extent during reperfusion. While MnSOD protein content was higher in IR group, activity returned to 95% of control. Nitric oxide is one of the biological molecules that can out compete MnSOD for superoxide and produce peroxynitrite. This process is faster than dismutation and led to the 10-fold higher production of nitrotyrosine after IR injury in adult with higher protection in senescent ones. 2D protein profiling revealed 140 mitochondrial proteins, 12 of them with significant changes after IR injury and 36 individual nitrotyrosine-modified proteins further identified by mass spectrometry. Linking these two groups, 5 proteins were altered after IR as well as nitrated, but only one showed massive nitration per lowering content of protein after IR injury in adult. Conclusions: Senescent cells have greater proportion of protein content, which might be modulated by several post-translational modifications. If these protein modifications are connected to functional consequences and protein-protein interactions are revealed, link may lead to the solution. Assume all together, dysfunctional proteostasis can play a causative role and restoration of protein homeostasis machinery is protective against aging and possibly age-related disorders. This work was supported by the project VEGA 1/0018/18 and by project 'Competence Center for Research and Development in the field of Diagnostics and Therapy of Oncological diseases', ITMS: 26220220153, co-financed from EU sources.

Keywords: aging heart, mitochondria, proteomics, redox state

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3 Towards Automatic Calibration of In-Line Machine Processes

Authors: David F. Nettleton, Elodie Bugnicourt, Christian Wasiak, Alejandro Rosales

Abstract:

In this presentation, preliminary results are given for the modeling and calibration of two different industrial winding MIMO (Multiple Input Multiple Output) processes using machine learning techniques. In contrast to previous approaches which have typically used ‘black-box’ linear statistical methods together with a definition of the mechanical behavior of the process, we use non-linear machine learning algorithms together with a ‘white-box’ rule induction technique to create a supervised model of the fitting error between the expected and real force measures. The final objective is to build a precise model of the winding process in order to control de-tension of the material being wound in the first case, and the friction of the material passing through the die, in the second case. Case 1, Tension Control of a Winding Process. A plastic web is unwound from a first reel, goes over a traction reel and is rewound on a third reel. The objectives are: (i) to train a model to predict the web tension and (ii) calibration to find the input values which result in a given tension. Case 2, Friction Force Control of a Micro-Pullwinding Process. A core+resin passes through a first die, then two winding units wind an outer layer around the core, and a final pass through a second die. The objectives are: (i) to train a model to predict the friction on die2; (ii) calibration to find the input values which result in a given friction on die2. Different machine learning approaches are tested to build models, Kernel Ridge Regression, Support Vector Regression (with a Radial Basis Function Kernel) and MPART (Rule Induction with continuous value as output). As a previous step, the MPART rule induction algorithm was used to build an explicative model of the error (the difference between expected and real friction on die2). The modeling of the error behavior using explicative rules is used to help improve the overall process model. Once the models are built, the inputs are calibrated by generating Gaussian random numbers for each input (taking into account its mean and standard deviation) and comparing the output to a target (desired) output until a closest fit is found. The results of empirical testing show that a high precision is obtained for the trained models and for the calibration process. The learning step is the slowest part of the process (max. 5 minutes for this data), but this can be done offline just once. The calibration step is much faster and in under one minute obtained a precision error of less than 1x10-3 for both outputs. To summarize, in the present work two processes have been modeled and calibrated. A fast processing time and high precision has been achieved, which can be further improved by using heuristics to guide the Gaussian calibration. Error behavior has been modeled to help improve the overall process understanding. This has relevance for the quick optimal set up of many different industrial processes which use a pull-winding type process to manufacture fibre reinforced plastic parts. Acknowledgements to the Openmind project which is funded by Horizon 2020 European Union funding for Research & Innovation, Grant Agreement number 680820

Keywords: data model, machine learning, industrial winding, calibration

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2 Experimental Study of the Behavior of Elongated Non-spherical Particles in Wall-Bounded Turbulent Flows

Authors: Manuel Alejandro Taborda Ceballos, Martin Sommerfeld

Abstract:

Transport phenomena and dispersion of non-spherical particle in turbulent flows are found everywhere in industrial application and processes. Powder handling, pollution control, pneumatic transport, particle separation are just some examples where the particle encountered are not only spherical. These types of multiphase flows are wall bounded and mostly highly turbulent. The particles found in these processes are rarely spherical but may have various shapes (e.g., fibers, and rods). Although research related to the behavior of regular non-spherical particles in turbulent flows has been carried out for many years, it is still necessary to refine models, especially near walls where the interaction fiber-wall changes completely its behavior. Imaging-based experimental studies on dispersed particle-laden flows have been applied for many decades for a detailed experimental analysis. These techniques have the advantages that they provide field information in two or three dimensions, but have a lower temporal resolution compared to point-wise techniques such as PDA (phase-Doppler anemometry) and derivations therefrom. The applied imaging techniques in dispersed two-phase flows are extensions from classical PIV (particle image velocimetry) and PTV (particle tracking velocimetry) and the main emphasis was simultaneous measurement of the velocity fields of both phases. In a similar way, such data should also provide adequate information for validating the proposed models. Available experimental studies on the behavior of non-spherical particles are uncommon and mostly based on planar light-sheet measurements. Especially for elongated non-spherical particles, however, three-dimensional measurements are needed to fully describe their motion and to provide sufficient information for validation of numerical computations. For further providing detailed experimental results allowing a validation of numerical calculations of non-spherical particle dispersion in turbulent flows, a water channel test facility was built around a horizontal closed water channel. Into this horizontal main flow, a small cross-jet laden with fiber-like particles was injected, which was also solely driven by gravity. The dispersion of the fibers was measured by applying imaging techniques based on a LED array for backlighting and high-speed cameras. For obtaining the fluid velocity fields, almost neutrally buoyant tracer was used. The discrimination between tracer and fibers was done based on image size which was also the basis to determine fiber orientation with respect to the inertial coordinate system. The synchronous measurement of fluid velocity and fiber properties also allow the collection of statistics of fiber orientation, velocity fields of tracer and fibers, the angular velocity of the fibers and the orientation between fiber and instantaneous relative velocity. Consequently, an experimental study the behavior of elongated non-spherical particles in wall bounded turbulent flows was achieved. The development of a comprehensive analysis was succeeded, especially near the wall region, where exists hydrodynamic wall interaction effects (e.g., collision or lubrication) and abrupt changes of particle rotational velocity. This allowed us to predict numerically afterwards the behavior of non-spherical particles within the frame of the Euler/Lagrange approach, where the particles are therein treated as “point-particles”.

Keywords: crossflow, non-spherical particles, particle tracking velocimetry, PIV

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1 The Role of a Biphasic Implant Based on a Bioactive Silk Fibroin for Osteochondral Tissue Regeneration

Authors: Lizeth Fuentes-Mera, Vanessa Perez-Silos, Nidia K. Moncada-Saucedo, Alejandro Garcia-Ruiz, Alberto Camacho, Jorge Lara-Arias, Ivan Marino-Martinez, Victor Romero-Diaz, Adolfo Soto-Dominguez, Humberto Rodriguez-Rocha, Hang Lin, Victor Pena-Martinez

Abstract:

Biphasic scaffolds in cartilage tissue engineering have been designed to influence not only the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone to promote the implant integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a biphasic scaffold based on the assembly of a cartilage phase constituted by fibroin biofunctionalized with bovine cartilage matrix; cellularized with differentiated pre-chondrocytes from adipose tissue stem cells (autologous) and well attached to a bone phase (bone bovine decellularized) to mimic the structure of the nature of native tissue and to promote the cartilage regeneration in a model of joint damage in pigs. Biphasic scaffolds were assembled by fibroin crystallization with methanol. The histological and ultrastructural architectures were evaluated by optical and scanning electron microscopy respectively. Mechanical tests were conducted to evaluate Young's modulus of the implant. For the biological evaluation, pre-chondrocytes were loaded onto the scaffolds and cellular adhesion, proliferation, and gene expression analysis of cartilage extracellular matrix components was performed. The scaffolds that were cellularized and matured for 10 days were implanted into critical 3 mm in diameter and 9-mm in depth osteochondral defects in a porcine model (n=4). Three treatments were applied per knee: Group 1: monophasic cellular scaffold (MS) (single chondral phase), group 2: biphasic scaffold, cellularized only in the chondral phase (BS1), group 3: BS cellularized in both bone and chondral phases (BS2). Simultaneously, a control without treatment was evaluated. After 4 weeks of surgery, integration and regeneration tissues were analyzed by x-rays, histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular biphasic composites exhibited Young's modulus of 805.01 kPa similar to native cartilage (400-800 kPa). In vitro biological studies revealed the chondroinductive ability of the biphasic implant, evidenced by an increase in sulfated glycosaminoglycan (GAGs) and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, In group 1, the defects were not reconstructed. In group 2 and 3 a good integration of the implant with the surrounding tissue was observed. Defects in group 2 were fulfilled by hyaline cartilage and normal bone. Group 3 defects showed fibrous repair tissue. In conclusion; our findings demonstrated the efficacy of biphasic and bioactive scaffold based on silk fibroin, which entwined chondroinductive features and biomechanical capability with appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.

Keywords: biphasic scaffold, extracellular cartilage matrix, silk fibroin, osteochondral tissue engineering

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