Search results for: S. Munoz

Authors: Camila Lozano-Vivas, Camila Quevedo-Villamil, Ingrid Munoz-Quijano, Diego Loaiza

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Colombia's limited oil reserves make the finding of new fields of extraction or the potentiate of the existing ones a more important task to do every day; the exploration projects that allow to have a better knowledge of the oil basins are essential. The upper Magdalena Valley basin - VSM, whose reserves are limited, has been one of the first basins for the exploration and production of hydrocarbons in Colombia. The Hondita and Lomagorda formations were deposited in the Late Cretaceous Middle Albian to the Coniacian and are characterized by being the hydrocarbon-generating rocks in the VSM basin oil system along with the Shale de Bambucá; therefore multiple studies have been made. In the oil industry, geochemical properties are used to understand the origin, migration, accumulation, and alteration of hydrocarbons and, in general, the evolution of the basin containing them. One of the most important parameters to understand this evolution is the formation temperature of the oil system. For this reason, a microthermometric study of fluid inclusions was carried out to recognize formation temperatures and to determine certain basic physicochemical variables, homogenization temperature, pressure, density and salinity of the fluid at the time of entrapment, providing evidence on the history of different events in different geological environments in the evolution of a sedimentary basin. Prior to this study, macroscopic and microscopic petrographic analyses of the samples collected in the field were performed. The results of the mentioned properties of the fluid inclusions in the different samples analyzed have salinities ranging from 20.22% to 26.37% eq. by weight NaCl, similar densities found in the ranges of 1.05 to 1.16 g/cc and an average homogenization temperature at 142.92°C, indicating that, at the time of their entanglement, the rock was in the window of generation of medium hydrocarbons –light with fragile characteristics of the rock that would make it useful to treat them as naturally fractured reservoirs.

Keywords: homogenization temperature, fluid inclusions, microthermometry, salinity

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Authors: Y. Vásquez, F. Reyes, P. Oyola, M. Rubio, J. Muñoz, E. Lissi

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In several cities in Chile, there is significant urban pollution, particularly in Santiago and in cities in the south where biomass is used as fuel in heating and cooking in a large proportion of homes. This has generated interest in knowing what factors can be modulated to control the level of pollution. In this project was conditioned and set up a photochemical chamber (14m3) equipped with gas monitors e.g. CO, NOX, O3, others and PM monitors e.g. dustrack, DMPS, Harvard impactors, etc. This volume could be exposed to UVA lamps, producing a spectrum similar to that generated by the sun. In this chamber, PM and gas emissions associated with biomass burning were studied in the presence and absence of radiation. From the comparative analysis of wood stove (eucalyptus globulus) and pellet (radiata pine), it can be concluded that, in the first approximation, 9-nitroanthracene, 4-nitropyrene, levoglucosan, water soluble potassium and CO present characteristics of the tracers. However, some of them show properties that interfere with this possibility. For example, levoglucosan is decomposed by radiation. The 9-nitroanthracene, 4-nitropyrene are emitted and formed under radiation. The 9-nitroanthracene has a vapor pressure that involves a partition involving the gas phase and particulate matter. From this analysis, it can be concluded that K+ is compound that meets the properties known to be tracer. The PM2.5 emission measured in the automatic pellet stove that was used in this thesis project was two orders of magnitude smaller than that registered by the manual wood stove. This has led to encouraging the use of pellet stoves in indoor heating, particularly in south-central Chile. However, it should be considered, while the use of pellet is not without problems, due to pellet stove generate high concentrations of Nitro-HAP's (secondary organic contaminants). In particular, 4-nitropyrene, compound of high toxicity, also primary and secondary particulate matter, associated with pellet burning produce a decrease in the size distribution of the PM, which leads to a depth penetration of the particles and their toxic components in the respiratory system.

Keywords: biomass burning, photochemical chamber, particulate matter, tracers

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Authors: Viviana Letelier, Ester Tarela, Bianca Lopez, Pedro Muñoz, Giacomo Moriconi

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The environmental impact has become a global concern during the last decades. Several alternatives have been proposed and studied to minimize this impact in different areas. The reuse of aggregates from old concretes to manufacture new ones not only can reduce this impact but is also a way to optimize the resource management. The effect of the origin of the reused aggregates from two different origin materials in recycled concrete pavement is studied here. Using the dosing applied by a pavement company, coarse aggregates in the 6.3-25 mm fraction are replaced by recycled aggregates with two different origins: old concrete pavements with similar origin strength to the one of the control concrete, and precast concrete pipes with smaller strengths than the one of the control concrete. The replacement percentages tested are 30%, 40% and 50% in both cases. The compressive strength tests are performed after 7, 14, 28 and 90 curing days, the flexural strength tests and the elasticity modulus tests after 28 and 90 curing days. Results show that the influence of the quality of the origin concrete in the mechanical properties of recycled concretes is not despicable. Concretes with up to a 50% of recycled aggregates from the concrete pavement have similar compressive strengths to the ones of the control concrete and slightly smaller flexural strengths that, however, in all cases exceed the minimum of 5MPa after 28 curing days stablished by the Chilean regulation for pavement concretes. On the other hand, concretes with recycled aggregates from precast concrete pipes show significantly lower compressive strengths after 28 curing days. The differences with the compressive strength of the control concrete increase with the percentage of replacement, reaching a 13% reduction when 50% of the aggregates are replaced. The flexural strength also suffers significant reductions that increase with the percentage of replacement, only obeying the Chilean regulation when 30% of the aggregates are recycled after 28 curing days. Nevertheless, after 90 curing days, all series obey the regulation requirements. Results show, not only the importance of the quality of the origin concrete, but also the significance of the curing days, that may allow the use of less quality recycled material without important strength losses.

Keywords: flexural strength of recycled concrete., mechanical properties of recycled concrete, recycled aggregates, recycled concrete pavements

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Authors: Ana Maria Igual-Munoz, Juan Gilabert, Marta Garcia, Alfredo Quijano-Lopez

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Nowadays, several lithium-ion battery technologies are being commercialized. These chemistries present different properties that make them more suitable for different purposes. However, comparative studies showing the advantages and disadvantages of different chemistries are incomplete or scarce. Different non-destructive techniques are currently being employed to detect how ageing affects the active materials of lithium-ion batteries (LIBs). For instance, electrochemical impedance spectroscopy (EIS) is one of the most employed ones. This technique allows the user to identify the variations on the different resistances present in LIBs. On the other hand, differential voltage analysis (DVA) has shown to be a powerful technique to detect the processes affecting the different capacities present in LIBs. This technique shows variations in the state of health (SOH) and the capacities for one or both electrodes depending on their chemistry. Finally, incremental capacity analysis (ICA) is a widely known technique for being capable of detecting phase equilibria. It reminds of the commonly used cyclic voltamperometry, as it allows detecting some reactions taking place in the electrodes. In these studies, a set of ageing procedures have been applied to commercial batteries of different chemistries (NCA, NMC, and LFP). Afterwards, results of EIS, DVA, and ICA have been used to correlate them with the processes affecting each cell. Ciclability, overpotential, and temperature cycling studies envisage how the charge-discharge rates, cut-off voltage, and operation temperatures affect each chemistry. These studies will serve battery pack manufacturers, as for common battery users, as they will determine the different conditions affecting cells for each of the chemistry. Taking this into account, each cell could be adjusted to the final purpose of the battery application. Last but not least, all the degradation parameters observed are focused to be integrated into degradation models in the future. This fact will allow the implementation of the widely known digital twins to the degradation in LIBs.

Keywords: lithium ion batteries, non-destructive analysis, different chemistries, ante-mortem studies, ICA, DVA, EIS

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Authors: Daniela N. Correa-Llantén, Sebastián A. Muñoz-Ibacache, Mathilde Maire, Jenny M. Blamey

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The biosynthesis of nanoparticles by microorganisms, on the contrary to chemical synthesis, is an environmentally-friendly process which has low energy requirements. In this investigation, we used the microorganism Geobacillus wiegelii, strain GWE1, an aerobic thermophile belonging to genus Geobacillus, isolated from a drying oven. This microorganism has the ability to reduce selenite evidenced by the change of color from colorless to red in the culture. Elemental analysis and composition of the particles were verified using transmission electron microscopy and energy-dispersive X-ray analysis. The nanoparticles have a defined spherical shape and a selenium elemental state. Previous experiments showed that the presence of the whole microorganism for the reduction of selenite was not necessary. The results strongly suggested that an intracellular NADPH/NADH-dependent reductase mediates selenium nanoparticles synthesis under aerobic conditions. The enzyme was purified and identified by mass spectroscopy MALDI-TOF TOF technique. The enzyme is a 1-pyrroline-5-carboxylate dehydrogenase. Histograms of nanoparticles sizes were obtained. Size distribution ranged from 40-160 nm, where 70% of nanoparticles have less than 100 nm in size. Spectroscopic analysis showed that the nanoparticles are composed of elemental selenium. To analyse the effect of pH in size and morphology of nanoparticles, the synthesis of them was carried out at different pHs (4.0, 5.0, 6.0, 7.0, 8.0). For thermostability studies samples were incubated at different temperatures (60, 80 and 100 ºC) for 1 h and 3 h. The size of all nanoparticles was less than 100 nm at pH 4.0; over 50% of nanoparticles have less than 100 nm at pH 5.0; at pH 6.0 and 8.0 over 90% of nanoparticles have less than 100 nm in size. At neutral pH (7.0) nanoparticles reach a size around 120 nm and only 20% of them were less than 100 nm. When looking at temperature effect, nanoparticles did not show a significant difference in size when they were incubated between 0 and 3 h at 60 ºC. Meanwhile at 80 °C the nanoparticles suspension lost its homogeneity. A change in size was observed from 0 h of incubation at 80ºC, observing a size range between 40-160 nm, with 20% of them over 100 nm. Meanwhile after 3 h of incubation at size range changed to 60-180 nm with 50% of them over 100 nm. At 100 °C the nanoparticles aggregate forming nanorod structures. In conclusion, these results indicate that is possible to modulate size and shape of biologically synthesized nanoparticles by modulating pH and temperature.

Keywords: genus Geobacillus, NADPH/NADH-dependent reductase, selenium nanoparticles, biosynthesis

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Authors: Laura Piedra-Muñoz, Ángeles Godoy-Durán, Emilio Galdeano-Gómez, Juan C. Pérez-Mesa

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The current agricultural system in some arid and semi-arid areas is not sustainable in the long term. In southeast Spain, groundwater is the main water source and is overexploited, while alternatives like desalination are still limited. The Water Plan for the Mediterranean Basins 2015-2020 indicates a global deficit of 73.42 hm3 and an overexploitation of the aquifers of 205.58hm3. In order to solve this serious problem, two major actions can be taken: increasing available water, and/or improving the efficiency of its use. This study focuses on the latter. The main aim of this study is to present the major factors related to water usage efficiency in farming. It focuses on Almería province, southeast Spain, one of the most arid areas of the country, and in particular on family farms as the main direct managers of water use in this zone. Many of these farms are among the most water efficient in Spanish agriculture, but this efficiency is not generalized throughout the sector. This work conducts a comprehensive assessment of water performance in this area, using on-farm water-use, structural, socio-economic and environmental information. Two statistical techniques are used: descriptive analysis and cluster analysis. Thus, two groups are identified: the least and the most efficient farms regarding water usage. By analyzing both the common characteristics within each group and the differences between the groups with a one-way ANOVA analysis, several conclusions can be reached. The main differences between the two clusters center on the extent to which innovation and new technologies are used in irrigation. The most water efficient farms are characterized by more educated farmers, a greater degree of innovation, new irrigation technology, specialized production and awareness of water issues and environmental sustainability. The research shows that better practices and policies can have a substantial impact on achieving a more sustainable and efficient use of water. The findings of this study can be extended to farms in similar arid and semi-arid areas and contribute to foster appropriate policies to improve the efficiency of water usage in the agricultural sector.

Keywords: cluster analysis, family farms, Spain, water-use efficiency

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Authors: Lohendy Munoz-Vargas, Stephen O. Opiyo, Rose Digianantonio, Michele L. Williams, Asela Wijeratne, Gregory Habing

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Non-typhoidal Salmonella enterica is a zoonotic pathogen with critical importance in animal and public health. The persistence of Salmonella on farms affects animal productivity and health, and represents a risk for food safety. The intestinal microbiota plays a fundamental role in the colonization and invasion of this ubiquitous microorganism. To overcome the colonization resistance imparted by the gut microbiome, Salmonella uses invasion strategies and the host inflammatory response to survive, proliferate, and establish infections with diverse clinical manifestations. Cattle serve as reservoirs of Salmonella, and periparturient cows have high prevalence of Salmonella shedding; however, to author`s best knowledge, little is known about the association between the gut microbiome and the onset of Salmonella shedding during the periparturient period. Thus, the objective of this study was to assess the association between changes in bacterial communities and the onset of Salmonella shedding in cattle approaching parturition. In a prospective cohort study, fecal samples from 98 dairy cows originating from four different farms were collected at four time points relative to calving (-3 wks, -1 wk, +1 wk, +3 wks). All 392 samples were cultured for Salmonella. Sequencing of the V4 region of the 16S rRNA gene using the Illumina platform was completed to evaluate the fecal microbiome in a selected sample subset. Analyses of microbial composition, diversity, and structure were performed according to time points, farm, and Salmonella onset status. Individual cow fecal microbiomes, predominated by Bacteroidetes, Firmicutes, Spirochaetes, and Proteobacteria phyla, significantly changed before and after parturition. Microbial communities from different farms were distinguishable based on multivariate analysis. Although there were significant differences in some bacterial taxa between Salmonella positive and negative samples, our results did not identify differences in the fecal microbial diversity or structure for cows with and without the onset of Salmonella shedding. These data suggest that determinants other than the significant changes in the fecal microbiome influence the periparturient onset of Salmonella shedding in dairy cattle.

Keywords: dairy cattle, microbiome, periparturient, Salmonella

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Authors: Renzo Munoz-Najar, Cristina Wong, Daniel De La Torre Ugarte

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The literature on price transmission usually deals with asymmetries related to different commodities and/or the short and long term. The role of domestic regional differences and the relationship with asymmetries within a country are usually left out. This paper looks at the asymmetry in the transmission of rice prices from the international price to the farm gate prices in four northern regions of Peru for the last period 2001-2016. These regions are San Martín, Piura, Lambayeque and La Libertad. The relevance of the study lies in its ability to assess the need for policies aimed at improving the competitiveness of the market and ensuring the benefit of producers. There are differences in planting and harvesting dates, as well as in geographic location that justify the hypothesis of the existence of differences in the price transition asymmetries between these regions. Those differences are due to at least three factors geography, infrastructure development, and distribution systems. For this, the Threshold Vector Error Correction Model and the Autoregressive Vector Model with Threshold are used. Both models, collect asymmetric effects in the price adjustments. In this way, it is sought to verify that farm prices react more to falls than increases in international prices due to the high bargaining power of intermediaries. The results of the investigation suggest that the transmission of prices is significant only for Lambayeque and La Libertad. Likewise, the asymmetry in the transmission of prices for these regions is checked. However, these results are not met for San Martin and Piura, the main rice producers nationwide. A significant price transmission is verified only in the Lambayeque and La Libertad regions. San Martin and Piura, in spite of being the main rice producing regions of Peru, do not present a significant transmission of international prices; a high degree of self-sufficient supply might be at the center of the logic for this result. An additional finding is the short-term adjustment with respect to international prices, it is higher in La Libertad compared to Lambayeque, which could be explained by the greater bargaining power of intermediaries in the last-mentioned region due to the greater technological development in the mills.

Keywords: asymmetric price transmission, rice prices, price transmission, regional economics

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Authors: Cristina Munoz-Shuguli, Francisco Rodriguez, Julio Bruna, M. Jose Galotto, Abel Guarda

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The microbial contamination in fruits due to the presence of fungal is the most important cause of their deterioration and loss. The development of active food packaging materials with antifungal properties has been proposed as an innovative strategy in order to prevent this problem. In this way, natural compounds as the essential oils or their derivatives, also called volatile compounds (VC), can be incorporated in the food packaging materials to control the fungal growth during fruit packaging. However, if the VC is incorporated directly in the packaging material, it is released very fast due to VC high volatility. For this reason, the formation of inclusion complexes through the encapsulation of VC into beta-cyclodextrin (β-CD) and their incorporation in package materials is an alternative to maintain an antifungal atmosphere around the packaged fruits for longer times. In this context, the aim of this work was to develop inclusion complexes based in β-CD and VC (β-CD:VC) for further application in the antifungal food packaging materials development. β-CD:VC inclusion complexes were obtained with two different molar ratios 2:1 and 1:1, through co-precipitation method. The entrapment efficiency of β-CD:VC as well the release of antifungal compound from inclusion complexes exposed to different relative humidity (25, 50, and 97 %) to headspace were determined by gaseous chromatography (GC). Also, thermal and antimicrobial properties of β-CD:VC were determined through thermogravimetric analysis (TGA) and antifungal assays against Botrytis cinerea, respectively. GC results showed that β-CD:VC 2:1 had a higher entrapment efficiency than β-CD:VC 1:1, with values of 75.5 ± 3.71 % and 59.6 ± 1.51 %, respectively. It was probably because during the synthesis of β-CD:VC 1:1, there was less molecular space to the movement of VC molecules. Furthermore, the release of VC from β-CD:VC was directly related with the relative humidity. High amount of VC was released when the inclusion complexes were exposed to high humidity, possibly due to the interactions between the water molecules and the β-CD hydrophilic wall. On the other hand, a better thermal stability of VC in inclusion complexes allowed to verify its effective encapsulation into β-CD. Finally, antimicrobial assays showed that the inclusion complexes had a high antifungal activity at very low concentrations. Therefore, the results obtained in this work allow suggesting the β-CD:VC inclusion complexes as potential candidates to the development of fruit antifungal packaging materials, which activity is relative humidity dependent.

Keywords: Botrytis cinerea, fruit packaging, headspace release, volatile compounds

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Authors: Gema Gómez-Casado, Alba Rodríguez-Muñoz, Virginia Mela-Rivas, Pallavi Kompella, Francisco José Tinahones-Madueña, Isabel Moreno-Indias, Almudena Ortega-Gómez

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Obesity is a high-priority health problem worldwide due to its high prevalence. The proportion of obese and overweight subjects in industrialized countries exceeds half of the population in most cases. Beyond the metabolic problem, obesity boosts inflammation levels in the organism. The gut microbiota, considered an organ by itself, controls a high variety of processes at a systemic level. In fact, the microbiota interacts closely with the immune system, being crucial in determining the maturation state of neutrophils, key effectors of the innate immune response. It is known that changes in the diet exert strong effects on the variety and activity of the gut microbiota. The effect that those changes have on the axis microbiota-immune response is an unexplored field. In this study, 10 patients with obesity (weight 114,3 ± 14,5Kg, BMI 40,47±3,66) followed a Mediterranean-hypocaloric diet for 3 months, reducing their initial weight by 12,71 ± 3%. A transplant of microbiota from these patients before and after the diet was performed into wild type “germ-free” mice (n=10/group), treated with antibiotics. Six weeks after the transplant, mice were euthanized, and the presence of cells from the innate immune system were analysed in different organs (bone marrow, blood, spleen, visceral adipose tissue, and intestine) by flow cytometry. No differences were observed in the number of myeloid cells in bone marrow, blood, spleen, or visceral adipose tissue of mice transplanted with patient’s microbiota before and after following the Mediterranean diet. However, the intestine of mice that received post-diet microbiota presented a marked decrease in the number of neutrophils (whose presence is associated with tissue inflammation), as well as macrophages. In line with these findings, intestine monocytes from mice with post-diet microbiota showed a less inflammatory profile (lower Ly6Gˡᵒʷ proportion of cells). These results point toward a decrease in the inflammatory state of the intestinal tissue, derived from changes in the gut microbiota, which occurred after a 3-month Mediterranean diet.

Keywords: obesity, nutrition, Mediterranean diet, gut microbiota, immune system

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Authors: Laura Rueda-Gensini, Jader Rodríguez, Juan C. Cruz, Carolina Munoz-Camargo

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There is an emerging interest in botanicals and plant extracts for medicinal practices due to their widely reported health benefits. A large variety of phytochemicals found in plants have been correlated with antioxidant, immunomodulatory, and analgesic properties, which makes plant-derived products promising candidates for modulating the progression and treatment of numerous diseases. Non-centrifugal cane sugar (NCS), in particular, has been known for its high antioxidant and nutritional value, but composition-wise variability due to changing environmental and processing conditions have considerably limited its use in the nutraceutical and biomedical fields. This work is therefore aimed at assessing the effect of thermal exposure during NCS production over its bioactive composition and, in turn, its therapeutic value. Accordingly, two modified dehydration methods are proposed that employ: (i) vacuum-aided evaporation, which reduces the necessary temperatures to dehydrate the sample, and (ii) window refractance evaporation, which reduces thermal exposure time. The biochemical composition of NCS produced under these two methods was compared to traditionally-produced NCS by estimating their total polyphenolic and protein content with Folin-Ciocalteu and Bradford assays, as well as identifying the major phenolic compounds in each sample via HPLC-coupled mass spectrometry. Their antioxidant activities were also compared as measured by their scavenging potential of ABTS and DPPH radicals. Results show that the two modified production methods enhance polyphenolic and protein yield in resulting NCS samples when compared to traditional production methods. In particular, reducing employed temperatures with vacuum-aided evaporation demonstrated to be superior at preserving polyphenolic compounds, as evidenced both in the total and individual polyphenol concentrations. However, antioxidant activities were not significantly different between these. Although additional studies should be performed to determine if the observed compositional differences affect other therapeutic activities (e.g., anti-inflammatory, analgesic, and immunoprotective), these results suggest that reducing thermal exposure holds great promise for the production of natural products with enhanced nutritional value.

Keywords: non-centrifugal cane sugar, polyphenolic compounds, thermal processing, antioxidant activity

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Authors: Emilio Galdeano-Gómez, Ángeles Godoy-Durán, Juan C. Pérez-Mesa, Laura Piedra-Muñoz

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Eco-efficiency is receiving ever-increasing interest as an indicator of sustainability, as it links environmental and economic performances in productive activities. In agriculture, these indicators and their determinants prove relevant due to the close relationships in this activity between the use of natural resources, which is generally limited, and the provision of basic goods to society. In this context, various analyses have focused on eco-efficiency by considering individual family farms as the basic production unit. However, not only must the measure of efficiency be taken into account, but also the existence of a series of factors which constitute socio-economic, political-institutional, and environmental determinants. Said factors have been studied to a lesser extent in the literature. The present work analyzes eco-efficiency at a micro level, focusing on small-scale family farms as the main decision-making units in horticulture in southeast Spain, a sector which represents about 30% of the fresh vegetables produced in the country and about 20% of those consumed in Europe. The objectives of this study are a) to obtain a series of eco-efficiency indicators by estimating several pressure ratios and economic value added in farming, b) to analyze the influence of specific social, economic and environmental variables on the aforementioned eco-efficiency indicators. The present work applies the method of Data Envelopment Analysis (DEA), which calculates different combinations of environmental pressures (water usage, phytosanitary contamination, waste management, etc.) and aggregate economic value. In a second stage, an analysis is conducted on the influence of the socio-economic and environmental characteristics of family farms on the eco-efficiency indicators, as endogeneous variables, through the use of truncated regression and bootstrapping techniques, following Simar-Wilson methodology. The results reveal considerable inefficiency in aspects such as waste management, while there is relatively little inefficiency in water usage and nitrogen balance. On the other hand, characteristics, such as product specialization, the adoption of quality certifications and belonging to a cooperative do have a positive impact on eco-efficiency. These results are deemed to be of interest to agri-food systems structured on small-scale producers, and they may prove useful to policy-makers as regards managing public environmental programs in agriculture.

Keywords: data envelopment analysis, eco-efficiency, family farms, horticulture, socioeconomic features

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Authors: Paola Leon, Hugo Garcia, Adan Leon, Samuel Munoz

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The enhanced oil recovery by steam injection was considered a process that only generated physical recovery mechanisms. However, there is evidence of the occurrence of a series of chemical reactions, which are called aquathermolysis, which generates hydrogen sulfide, carbon dioxide, methane, and lower molecular weight hydrocarbons. These reactions can be favored by the addition of a catalyst during steam injection; in this way, it is possible to generate the original oil in situ upgrading through the production increase of molecules of lower molecular weight. This additional effect could increase the oil recovery factor and reduce costs in transport and refining stages. Therefore, this research has focused on the experimental evaluation of the catalytic aquathermolysis on a Colombian heavy oil with 12,8°API. The effects of three different catalysts, reaction time, and temperature were evaluated in a batch microreactor. The changes in the Colombian heavy oil were quantified through nuclear magnetic resonance 1H-NMR. The relaxation times interpretation and the absorption intensity allowed to identify the distribution of the functional groups in the base oil and upgraded oils. Additionally, the average number of aliphatic carbons in alkyl chains, the number of substituted rings, and the aromaticity factor were established as average structural parameters in order to simplify the samples' compositional analysis. The first experimental stage proved that each catalyst develops a different reaction mechanism. The aromaticity factor has an increasing order of the salts used: Mo > Fe > Ni. However, the upgraded oil obtained with iron naphthenate tends to form a higher content of mono-aromatic and lower content of poly-aromatic compounds. On the other hand, the results obtained from the second phase of experiments suggest that the upgraded oils have a smaller difference in the length of alkyl chains in the range of 240º to 270°C. This parameter has lower values at 300°C, which indicates that the alkylation or cleavage reactions of alkyl chains govern at higher reaction temperatures. The presence of condensation reactions is supported by the behavior of the aromaticity factor and the bridge carbons production between aromatic rings (RCH₂). Finally, it is observed that there is a greater dispersion in the aliphatic hydrogens, which indicates that the alkyl chains have a greater reactivity compared to the aromatic structures.

Keywords: catalyst, upgrading, aquathermolysis, steam

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Authors: Lisseth Rojas Barreto, Carlos Muñoz Hernández

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The COVID-19 pandemic led to many abrupt changes, including a prolonged lockdown, which brought about work and personal challenges to the population worldwide. Among the most affected populations are women who are workers and housewives at the same time, and especially those who are also parenting. These women were faced with the challenge to perform their usual varied roles during the lockdown from the same physical space, which inevitably had strong repercussions for each of them. This paper will present some results of a research study whose main objective was to examine the possible effects that the COVID-19 pandemic lockdown may have caused in the work, social, family, and personal environments of female English teachers who are also housewives and, by extension in the teaching and learning processes that they lead. Participants included five female English language teachers of a public foreign language school, they are all married, and two of them have children. Similarly, we examined some of the coping strategies these teachers used to tackle the pandemic-related challenges in their different roles, especially those used for their language teaching role; coping strategies are understood as a repertoire of behaviors in response to incidents that can be stressful for the subject, possible challenging events or situations that involve emotions with behaviors and decision-making of people which are used in order to find a meaning or positive result (Lazarus &Folkman, 1986) Following a qualitative-case study design, we gathered the data through a survey and a focus group interview with the participant teachers who work at a public language school in southern Colombia. Preliminary findings indicate that the circumstances that emerged as a result of the pandemic lockdown affected the participants in different ways, including financial, personal, family, health, and work-related issues. Among the strategies that participants found valuable to deal with the novel circumstances, we can highlight the reorganization of the household and work tasks and the increased awareness of time management for the household, work, and leisure. Additionally, we were able to evidence that the participants faced the circumstances with a positive view. Finally, in order to cope with their teaching duties, some participants acknowledged their lack of computer or technology literacy in order to deliver their classes online, which made them find support from their students or more knowledgeable peers to cope with it. Others indicated that they used strategies such as self-learning in order to get acquainted and be able to use the different technological tools and web-based platforms available.

Keywords: coping strategies, language teaching, female teachers, pandemic lockdown

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Authors: Mindaye Teshome, Evaldo Muñoz Braz, Carlos M. M. Eleto Torres, Patricia Mattos

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Sustainable forest management planning requires up-to-date information on the structure, standing volume, biomass, and growth rate of trees from a given forest. This kind of information is lacking in many forests in Ethiopia. The objective of this study was to quantify the population structure, diameter growth rate, and standing volume of wood from Juniperus procera trees in the Chilimo forest. A total of 163 sample plots were set up in the forest to collect the relevant vegetation data. Growth ring measurements were conducted on stem disc samples collected from 12 J. procera trees. Diameter and height measurements were recorded from a total of 1399 individual trees with dbh ≥ 2 cm. The growth rate, maximum current and mean annual increments, minimum logging diameter, and cutting cycle were estimated, and alternative cutting cycles were established. Using these data, the harvestable volume of wood was projected by alternating four minimum logging diameters and five cutting cycles following the stand table projection method. The results show that J. procera trees have an average density of 183 stems ha⁻¹, a total basal area of 12.1 m² ha⁻¹, and a standing volume of 98.9 m³ ha⁻¹. The mean annual diameter growth ranges between 0.50 and 0.65 cm year⁻¹ with an overall mean of 0.59 cm year⁻¹. The population of J. procera tree followed a reverse J-shape diameter distribution pattern. The maximum current annual increment in volume (CAI) occurred at around 49 years when trees reached 30 cm in diameter. Trees showed the maximum mean annual increment in volume (MAI) around 91 years, with a diameter size of 50 cm. The simulation analysis revealed that 40 cm MLD and a 15-year cutting cycle are the best minimum logging diameter and cutting cycle. This combination showed the largest harvestable volume of wood potential, volume increments, and a 35% recovery of the initially harvested volume. It is concluded that the forest is well stocked and has a large amount of harvestable volume of wood from J. procera trees. This will enable the country to partly meet the national wood demand through domestic wood production. The use of the current population structure and diameter growth data from tree ring analysis enables the exact prediction of the harvestable volume of wood. The developed model supplied an idea about the productivity of the J. procera tree population and enables policymakers to develop specific management criteria for wood harvesting.

Keywords: logging, growth model, cutting cycle, minimum logging diameter

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Authors: Henrry Vicente Rojas Asuero, Holger Manuel Benavides Muñoz

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Approximately 80% of the energy consumed worldwide is generated from fossil sources, which are responsible for the emission of a large volume of greenhouse gases. For this reason, the global trend, at present, is the widespread use of energy produced from renewable sources. This seeks safety and diversification of energy supply, based on social cohesion, economic feasibility and environmental protection. In this scenario, small hydropower systems (P ≤ 10MW) stand out due to their high efficiency, economic competitiveness and low environmental impact. Small hydropower systems, along with wind and solar energy, are expected to represent a significant percentage of the world's energy matrix in the near term. Among the various technologies present in the state of the art, relating to small hydropower systems, is the Gravitational Water Vortex Power Plant, a recent technology that excels because of its versatility of operation, since it can operate with jumps in the range of 0.70 m-2.00 m and flow rates from 1 m3/s to 20 m3/s. Its operating system is based on the utilization of the energy of rotation contained within a large water vortex artificially induced. This paper presents the study and experimental design of an optimal hydraulic structure with the capacity to induce the artificial formation of a gravitational water vortex trough a system of easy application and high efficiency, able to operate in conditions of very low head and minimum flow. The proposed structure consists of a channel, with variable base, vortex inductor, tangential flow generator, coupled to a circular tank with a conical transition bottom hole. In the laboratory test, the angular velocity of the water vortex was related to the geometric characteristics of the inductor channel, as well as the influence of the conical transition bottom hole on the physical characteristics of the water vortex. The results show angular velocity values of greater magnitude as a function of depth, in addition the presence of the conical transition in the bottom hole of the circular tank improves the water vortex formation conditions while increasing the angular velocity values. Thus, the proposed system is a sustainable solution for the energy supply of rural areas near to watercourses.

Keywords: experimental model, gravitational water vortex power plant, renewable energy, small hydropower

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Authors: Rocío González-Sánchez, Sara Alonso-Muñoz

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Food waste implies an inefficient management of the final stages in the food supply chain. Referring to Sustainable Development Goals (SDGs) by United Nations, the SDG 12.3 proposes to halve per capita food waste at the retail and consumer level and to reduce food losses. In the linear system, food waste is disposed and, to a lesser extent, recovery or reused after consumption. With the negative effect on stocks, the current food consumption system is based on ‘produce, take and dispose’ which put huge pressure on raw materials and energy resources. Therefore, greater focus on the circular management of food waste will mitigate the environmental, economic, and social impact, following a Triple Bottom Line (TBL) approach and consequently the SDGs fulfilment. A mixed methodology is used. A total sample of 311 publications from Web of Science database were retrieved. Firstly, it is performed a bibliometric analysis by SciMat and VOSviewer software to visualise scientific maps about co-occurrence analysis of keywords and co-citation analysis of journals. This allows for the understanding of the knowledge structure about this field, and to detect research issues. Secondly, a systematic literature review is conducted regarding the most influential articles in years 2020 and 2021, coinciding with the most representative period under study. Thirdly, to support the development of this field it is proposed an agenda according to the research gaps identified about circular economy and food waste management. Results reveal that the main topics are related to waste valorisation, the application of waste-to-energy circular model and the anaerobic digestion process towards fossil fuels replacement. It is underlined that the use of food as a source of clean energy is receiving greater attention in the literature. There is a lack of studies about stakeholders’ awareness and training. In addition, available data would facilitate the implementation of circular principles for food waste recovery, management, and valorisation. The research agenda suggests that circularity networks with suppliers and customers need to be deepened. Technological tools for the implementation of sustainable business models, and greater emphasis on social aspects through educational campaigns are also required. This paper contributes on the application of circularity to food waste management by abandoning inefficient linear models. Shedding light about trending topics in the field guiding to scholars for future research opportunities.

Keywords: bibliometric analysis, circular economy, food waste management, future research lines

Procedia PDF Downloads 74

Authors: Willian C. De Brito, Hernan D. C. Munoz, Erlan V. C. Carvalho, Helder L. C. De Oliveira

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In order to improve commute time for small distance trips and relieve large cities traffic, a new transport category has been the subject of research and new designs worldwide. The air taxi travel market promises to change the way people live and commute by using the concept of vehicles with the ability to take-off and land vertically and to provide passenger’s transport equivalent to a car, with mobility within large cities and between cities. Today’s civil air transport remains costly and accounts for 2% of the man-made CO₂ emissions. Taking advantage of this scenario, many companies have developed their own Vertical Take Off and Landing (VTOL) design, seeking to meet comfort, safety, low cost and flight time requirements in a sustainable way. Thus, the use of green power supplies, especially batteries, and fully electric power plants is the most common choice for these arising aircrafts. However, it is still a challenge finding a feasible way to handle with the use of batteries rather than conventional petroleum-based fuels. The batteries are heavy and have an energy density still below from those of gasoline, diesel or kerosene. Therefore, despite all the clear advantages, all electric aircrafts (AEA) still have low flight autonomy and high operational cost, since the batteries must be recharged or replaced. In this sense, this paper addresses a way to optimize the energy consumption in a typical mission of an aerial taxi aircraft. The approach and landing procedure was chosen to be the subject of an optimization genetic algorithm, while final programming can be adapted for take-off and flight level changes as well. A real tilt rotor aircraft with fully electric power plant data was used to fit the derived dynamic equations of motion. Although a tilt rotor design is used as a proof of concept, it is possible to change the optimization to be applied for other design concepts, even those with independent motors for hover and cruise flight phases. For a given trajectory, the best set of control variables are calculated to provide the time history response for aircraft´s attitude, rotors RPM and thrust direction (or vertical and horizontal thrust, for independent motors designs) that, if followed, results in the minimum electric power consumption through that landing path. Safety, comfort and design constraints are assumed to give representativeness to the solution. Results are highly dependent on these constraints. For the tested cases, performance improvement ranged from 5 to 10% changing initial airspeed, altitude, flight path angle, and attitude.

Keywords: air taxi travel, all electric aircraft, batteries, energy consumption, genetic algorithm, landing performance, optimization, performance improvement, tilt rotor, VTOL design

Procedia PDF Downloads 84

Authors: Álvaro Ramírez Vidal, Martín Muñoz Morales, Francisco Jesús Fernández Morales, Luis Rodríguez Romero, José Villaseñor Camacho, Javier Llanos López

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In recent times, environmental problems have led to the extensive use of biological systems to solve them. Among the different types of biological systems, the use of plants such as aquatic macrophytes in constructed wetlands and terrestrial plant species for treating polluted soils and sludge has gained importance. Though the use of constructed wetlands for wastewater treatment is a well-researched domain, the slowness of pollutant degradation and high biomass production pose some challenges. Plants used in CW participate in different mechanisms for the capture and degradation of pollutants that also can retain some pharmaceutical and personal care products (PPCPs) that are very persistent in the environment. Thus, these systems present advantages in line with the guidelines published for the transition towards friendly and ecological procedures as they are environmentally friendly systems, consume low energy, or capture atmospheric CO₂. However, the use of CW presents some drawbacks, as the slowness of pollutant degradation or the production of important amounts of plant biomass, which need to be harvested and managed periodically. Taking this opportunity in mind, it is important to highlight that this residual biomass (of lignocellulosic nature) could be used as the feedstock for the generation of carbonaceous materials using thermochemical transformations such as slow pyrolysis or hydrothermal carbonization to produce high-value biomass-derived carbons through sustainable processes as adsorbents, catalysts…, thereby improving the circular carbon economy. Thus, this work carried out the analysis of some PPCPs commonly found in urban wastewater, as salicylic acid or ibuprofen, to evaluate the remediation carried out for the Phragmites Australis. Then, after the harvesting, this biomass can be used to synthesize electrodes through hydrothermal carbonization (HTC) and produce high-value biomass-derived carbons with electrocatalytic activity to remove heavy metals and persistent pollutants, promoting circular economy concepts. To do this, it was chosen biomass derived from the natural environment in high environmental risk as the Daimiel Wetlands National Park in the center of Spain, and the rest of the biomass developed in a CW specifically designed to remove pollutants. The research emphasizes the impact of the composition of the biomass waste and the synthetic parameters applied during HTC on the electrocatalytic activity. Additionally, this parameter can be related to the physicochemical properties, as porosity, surface functionalization, conductivity, and mass transfer of the electrodes lytic inks. Data revealed that carbon materials synthesized have good surface properties (good conductivities and high specific surface area) that enhance the electro-oxidants generated and promote the removal of PPCPs and the chemical oxygen demand of polluted waters.

Keywords: constructed wetlands, carbon materials, heavy metals, pharmaceutical and personal care products, hydrothermal carbonization

Procedia PDF Downloads 49

Authors: Javier E. García-Gallo, Nelson J. Fonseca-Ruiz, John F. Duitama-Munoz

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Sepsis is a syndrome that occurs with physiological and biochemical abnormalities induced by severe infection and carries a high mortality and morbidity, therefore the severity of its condition must be interpreted quickly. After patient admission in an intensive care unit (ICU), it is necessary to synthesize the large volume of information that is collected from patients in a value that represents the severity of their condition. Traditional severity of illness scores seeks to be applicable to all patient populations, and usually assess in-hospital mortality. However, the use of machine learning techniques and the data of a population that shares a common characteristic could lead to the development of customized mortality prediction scores with better performance. This study presents the development of a score for the one-year mortality prediction of the patients that are admitted to an ICU with a sepsis diagnosis. 5650 ICU admissions extracted from the MIMICIII database were evaluated, divided into two groups: 70% to develop the score and 30% to validate it. Comorbidities, demographics and clinical information of the first 24 hours after the ICU admission were used to develop a mortality prediction score. LASSO (least absolute shrinkage and selection operator) and SGB (Stochastic Gradient Boosting) variable importance methodologies were used to select the set of variables that make up the developed score; each of this variables was dichotomized and a cut-off point that divides the population into two groups with different mean mortalities was found; if the patient is in the group that presents a higher mortality a one is assigned to the particular variable, otherwise a zero is assigned. These binary variables are used in a logistic regression (LR) model, and its coefficients were rounded to the nearest integer. The resulting integers are the point values that make up the score when multiplied with each binary variables and summed. The one-year mortality probability was estimated using the score as the only variable in a LR model. Predictive power of the score, was evaluated using the 1695 admissions of the validation subset obtaining an area under the receiver operating characteristic curve of 0.7528, which outperforms the results obtained with Sequential Organ Failure Assessment (SOFA), Oxford Acute Severity of Illness Score (OASIS) and Simplified Acute Physiology Score II (SAPSII) scores on the same validation subset. Observed and predicted mortality rates within estimated probabilities deciles were compared graphically and found to be similar, indicating that the risk estimate obtained with the score is close to the observed mortality, it is also observed that the number of events (deaths) is indeed increasing as the outcome go from the decile with the lowest probabilities to the decile with the highest probabilities. Sepsis is a syndrome that carries a high mortality, 43.3% for the patients included in this study; therefore, tools that help clinicians to quickly and accurately predict a worse prognosis are needed. This work demonstrates the importance of customization of mortality prediction scores since the developed score provides better performance than traditional scoring systems.

Keywords: intensive care, logistic regression model, mortality prediction, sepsis, severity of illness, stochastic gradient boosting

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Authors: Óscar Muñoz Garrigós, José Manuel Bernabeu Aubán

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Time-sensitive services are the base of the cloud services industry. Keeping low service saturation is essential for controlling response time. All auto-scalable services make use of reactive auto-scaling. However, reactive auto-scaling has few in-depth studies. This presentation shows a model for reactive auto-scaling methodologies with a MAPE-k architecture. Queuing theory can compute different properties of static services but lacks some parameters related to the transition between models. Our model uses queuing theory parameters to relate the transition between models. It associates MAPE-k related times, the sampling frequency, the cooldown period, the number of requests that an instance can handle per unit of time, the number of incoming requests at a time instant, and a function that describes the acceleration in the service's ability to handle more requests. This model is later used as a solution to horizontally auto-scale time-sensitive services composed of microservices, reevaluating the model’s parameters periodically to allocate resources. The solution requires limiting the acceleration of the growth in the number of incoming requests to keep a constrained response time. Business benefits determine such limits. The solution can add a dynamic number of instances and remains valid under different system sizes. The study includes performance recommendations to improve results according to the incoming load shape and business benefits. The exposed methodology is tested in a simulation. The simulator contains a load generator and a service composed of two microservices, where the frontend microservice depends on a backend microservice with a 1:1 request relation ratio. A common request takes 2.3 seconds to be computed by the service and is discarded if it takes more than 7 seconds. Both microservices contain a load balancer that assigns requests to the less loaded instance and preemptively discards requests if they are not finished in time to prevent resource saturation. When load decreases, instances with lower load are kept in the backlog where no more requests are assigned. If the load grows and an instance in the backlog is required, it returns to the running state, but if it finishes the computation of all requests and is no longer required, it is permanently deallocated. A few load patterns are required to represent the worst-case scenario for reactive systems: the following scenarios test response times, resource consumption and business costs. The first scenario is a burst-load scenario. All methodologies will discard requests if the rapidness of the burst is high enough. This scenario focuses on the number of discarded requests and the variance of the response time. The second scenario contains sudden load drops followed by bursts to observe how the methodology behaves when releasing resources that are lately required. The third scenario contains diverse growth accelerations in the number of incoming requests to observe how approaches that add a different number of instances can handle the load with less business cost. The exposed methodology is compared against a multiple threshold CPU methodology allocating/deallocating 10 or 20 instances, outperforming the competitor in all studied metrics.

Keywords: reactive auto-scaling, auto-scaling, microservices, cloud computing

Procedia PDF Downloads 66

Authors: Damien Mouratille, Antonio R. Hidalgo-Muñoz, Nadine Matton, Yves Rouillard, Mickael Causse, Radouane El Yagoubi

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Background: The monitoring of the physiological activity related to mental workload (MW) on pilots will be useful to improve aviation safety by anticipating human performance degradation. The electrocardiogram (ECG) can reveal MW fluctuations due to either cognitive workload or/and emotional state since this measure exhibits autonomic nervous system modulations. Arguably, heart rate (HR) is one of its most intuitive and reliable parameters. It would be particularly interesting to analyze the interaction between cognitive requirements and emotion in ecologic sets such as a flight simulator. This study aims to explore by means of HR the relation between cognitive demands and emotional activation. Presumably, the effects of cognition and emotion overloads are not necessarily cumulative. Methodology: Eight healthy volunteers in possession of the Private Pilot License were recruited (male; 20.8±3.2 years). ECG signal was recorded along the whole experiment by placing two electrodes on the clavicle and left pectoral of the participants. The HR was computed within 4 minutes segments. NASA-TLX and Big Five inventories were used to assess subjective workload and to consider the influence of individual personality differences. The experiment consisted in completing two dual-tasks of approximately 30 minutes of duration into a flight simulator AL50. Each dual-task required the simultaneous accomplishment of both a pre-established flight plan and an additional task based on target stimulus discrimination inserted between Air Traffic Control instructions. This secondary task allowed us to vary the cognitive workload from low (LC) to high (HC) levels, by combining auditory and visual numerical stimuli to respond to meeting specific criteria. Regarding emotional condition, the two dual-tasks were designed to assure analogous difficulty in terms of solicited cognitive demands. The former was realized by the pilot alone, i.e. Low Arousal (LA) condition. In contrast, the latter generates a high arousal (HA), since the pilot was supervised by two evaluators, filmed and involved into a mock competition with the rest of the participants. Results: Performance for the secondary task showed significant faster reaction times (RT) for HA compared to LA condition (p=.003). Moreover, faster RT was found for LC compared to HC (p < .001) condition. No interaction was found. Concerning HR measure, despite the lack of main effects an interaction between emotion and cognition is evidenced (p=.028). Post hoc analysis showed smaller HR for HA compared to LA condition only for LC (p=.049). Conclusion. The control of an aircraft is a very complex task including strong cognitive demands and depends on the emotional state of pilots. According to the behavioral data, the experimental set has permitted to generate satisfactorily different emotional and cognitive levels. As suggested by the interaction found in HR measure, these two factors do not seem to have a cumulative impact on the sympathetic nervous system. Apparently, low cognitive workload makes pilots more sensitive to emotional variations. These results hint the independency between data processing and emotional regulation. Further physiological data are necessary to confirm and disentangle this relation. This procedure may be useful for monitoring objectively pilot’s mental workload.

Keywords: cognitive demands, emotion, flight simulator, heart rate, mental workload

Procedia PDF Downloads 247

Authors: Emilio Galdeano-Gómez, Juan C. Pérez-Mesa, Laura Piedra-Muñoz, María C. García-Barranco, Jesús Hernández-Rubio

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The situation in which firms engage in simultaneous cooperation and competition with each other is a phenomenon known as co-opetition. This scenario has received increasing attention in business economics and management analyses. In the domain of supply chain networks and for small and medium-sized enterprises, SMEs, these strategies are of greater relevance given the complex environment of globalization and competition in open markets. These firms face greater challenges regarding technology and access to specific resources due to their limited capabilities and limited market presence. Consequently, alliances and collaborations with both buyers and suppliers prove to be key elements in overcoming these constraints. However, rivalry and competition are also regarded as major factors in successful internationalization processes, as they are drivers for firms to attain a greater degree of specialization and to improve efficiency, for example enabling them to allocate scarce resources optimally and providing incentives for innovation and entrepreneurship. The present work aims to contribute to the literature on SMEs’ internationalization strategies. The sample is constituted by a panel data of marketing firms from the Andalusian food sector and a multivariate regression analysis is developed, measuring variables of co-opetition and international activity. The hierarchical regression equations method has been followed, thus resulting in three estimated models: the first one excluding the variables indicative of channel type, while the latter two include the international retailer chain and wholesaler variable. The findings show that the combination of several factors leads to a complex scenario of inter-organizational relationships of cooperation and competition. In supply chain management analyses, these relationships tend to be classified as either buyer-supplier (vertical level) or supplier-supplier relationships (horizontal level). Several buyers and suppliers tend to participate in supply chain networks, and in which the form of governance (hierarchical and non-hierarchical) influences cooperation and competition strategies. For instance, due to their market power and/or their closeness to the end consumer, some buyers (e.g. large retailers in food markets) can exert an influence on the selection and interaction of several of their intermediate suppliers, thus endowing certain networks in the supply chain with greater stability. This hierarchical influence may in turn allow these suppliers to develop their capabilities (e.g. specialization) to a greater extent. On the other hand, for those suppliers that are outside these networks, this environment of hierarchy, characterized by a “hub firm” or “channel master”, may provide an incentive for developing their co-opetition relationships. These results prove that the analyzed firms have experienced considerable growth in sales to new foreign markets, mainly in Europe, dealing with large retail chains and wholesalers as main buyers. This supply industry is predominantly made up of numerous SMEs, which has implied a certain disadvantage when dealing with the buyers, as negotiations have traditionally been held on an individual basis and in the face of high competition among suppliers. Over recent years, however, cooperation among these marketing firms has become more common, for example regarding R&D, promotion, scheduling of production and sales.

Keywords: co-petition networks, international supply chain, maketing agrifood firms, SMEs strategies

Procedia PDF Downloads 50

Authors: Adriana Muñoz-Talavera, Ana Rosa Rincón-Sánchez, Abraham Escobedo-Moratilla, María Cristina Islas-Carbajal, Miguel Ángel Gómez-Lim

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The highest rates of diabetes incidence and prevalence worldwide will increase the number of diabetic patients requiring insulin or insulin analogues. Then, current production systems would not be sufficient to meet the future market demands. Therefore, developing efficient expression systems for insulin and insulin analogues are needed. In addition, insulin analogues with better pharmacokinetics and pharmacodynamics properties and without mitogenic potential will be required. SCI-57 (single chain insulin-57) is an insulin analogue having 10 times greater affinity to the insulin receptor, higher resistance to thermal degradation than insulin, native mitogenicity and biological effect. Plants as expression platforms have been used to produce recombinant proteins because of their advantages such as cost-effectiveness, posttranslational modifications, absence of human pathogens and high quality. Immunoglobulin production with a yield of 50% has been achieved by transient expression in Nicotiana benthamiana (Nb). The aim of this study is to produce SCI-57 by transient expression in Nb. Methodology: DNA sequence encoding SCI-57 was cloned in pICH31070. This construction was introduced into Agrobacterium tumefaciens by electroporation. The resulting strain was used to infiltrate leaves of Nb. In order to isolate SCI-57, leaves from transformed plants were incubated 3 hours with the extraction buffer therefore filtrated to remove solid material. The resultant protein solution was subjected to anion exchange chromatography on an FPLC system and ultrafiltration to purify SCI-57. Detection of SCI-57 was made by electrophoresis pattern (SDS-PAGE). Protein band was digested with trypsin and the peptides were analyzed by Liquid chromatography tandem-mass spectrometry (LC-MS/MS). A purified protein sample (20µM) was analyzed by ESI-Q-TOF-MS to obtain the ionization pattern and the exact molecular weight determination. Chromatography pattern and impurities detection were performed using RP-HPLC using recombinant insulin as standard. The identity of the SCI-57 was confirmed by anti-insulin ELISA. The total soluble protein concentration was quantified by Bradford assay. Results: The expression cassette was verified by restriction mapping (5393 bp fragment). The SDS-PAGE of crude leaf extract (CLE) of transformed plants, revealed a protein of about 6.4 kDa, non-present in CLE of untransformed plants. The LC-MS/MS results displayed one peptide with a high score that matches SCI-57 amino acid sequence in the sample, confirming the identity of SCI-57. From the purified SCI-57 sample (PSCI-57) the most intense charge state was 1069 m/z (+6) on the displayed ionization pattern corresponding to the molecular weight of SCI-57 (6412.6554 Da). The RP-HPLC of the PSCI-57 shows the presence of a peak with similar retention time (rt) and UV spectroscopic profile to the insulin standard (SCI-57 rt=12.96 and insulin rt=12.70 min). The collected SCI-57 peak had ELISA signal. The total protein amount in CLE from transformed plants was higher compared to untransformed plants. Conclusions: Our results suggest the feasibility to produce insulin analogue SCI-57 by transient expression in Nicotiana benthamiana. Further work is being undertaken to evaluate the biological activity by glucose uptake by insulin-sensitive and insulin-resistant murine and human cultured adipocytes.

Keywords: insulin analogue, mass spectrometry, Nicotiana benthamiana, transient expression

Procedia PDF Downloads 319

Authors: Kalaumari Mayoral-Peña, Ana I. Montejano-Montelongo, Josué Reyes-Muñoz, Gonzalo A. Ortiz-Mancilla, Mayrin Rodríguez-Cruz, Víctor Hernández-Villalobos, Jesús A. Guzmán-López, Santiago García-Jacobo, Iván Licona-Vázquez, Grisel Fierros-Romero, Rosario Flores-Vallejo

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The group B-lactamic antibiotics include some of the most frequently used small drug molecules against bacterial infections. Nevertheless, an alarming decrease in their efficacy has been reported due to the emergence of antibiotic-resistant bacteria. Infections caused by bacteria expressing extended Spectrum B-lactamases (ESBLs) are difficult to treat and account for higher morbidity and mortality rates, delayed recovery, and high economic burden. According to the Global Report on Antimicrobial Resistance Surveillance, it is estimated that mortality due to resistant bacteria will ascend to 10 million cases per year worldwide. These facts highlight the importance of developing low-cost and readily accessible detection methods of drug-resistant ESBLs bacteria to prevent their spread and promote accurate and fast diagnosis. Bacterial detection is commonly done using molecular diagnostic techniques, where PCR stands out for its high performance. However, this technique requires specialized equipment not available everywhere, is time-consuming, and has a high cost. Loop-Mediated Isothermal Amplification (LAMP) is an alternative technique that works at a constant temperature, significantly decreasing the equipment cost. It yields double-stranded DNA of several lengths with repetitions of the target DNA sequence as a product. Although positive and negative results from LAMP can be discriminated by colorimetry, fluorescence, and turbidity, there is still a large room for improvement in the point-of-care implementation. DNA origami is a technique that allows the formation of 3D nanometric structures by folding a large single-stranded DNA (scaffold) into a determined shape with the help of short DNA sequences (staples), which hybridize with the scaffold. This research aimed to generate DNA origami structures using LAMP products as scaffolds to improve the sensitivity to detect ESBLs in point-of-care diagnosis. For this study, the coding sequence of the CTM-X-15 ESBL of E. coli was used to generate the LAMP products. The set of LAMP primers were designed using PrimerExplorerV5. As a result, a target sequence of 200 nucleotides from CTM-X-15 ESBL was obtained. Afterward, eight different DNA origami structures were designed using the target sequence in the SDCadnano and analyzed with CanDo to evaluate the stability of the 3D structures. The designs were constructed minimizing the total number of staples to reduce costs and complexity for point-of-care applications. After analyzing the DNA origami designs, two structures were selected. The first one was a zig-zag flat structure, while the second one was a wall-like shape. Given the sequence repetitions in the scaffold sequence, both were able to be assembled with only 6 different staples each one, ranging between 18 to 80 nucleotides. Simulations of both structures were performed using scaffolds of different sizes yielding stable structures in all the cases. The generation of the LAMP products were tested by colorimetry and electrophoresis. The formation of the DNA structures was analyzed using electrophoresis and colorimetry. The modeling of novel detection methods through bioinformatics tools allows reliable control and prediction of results. To our knowledge, this is the first study that uses LAMP products and DNA-origami in combination to delect ESBL-producing bacterial strains, which represent a promising methodology for diagnosis in the point-of-care.

Keywords: beta-lactamases, antibiotic resistance, DNA origami, isothermal amplification, LAMP technique, molecular diagnosis

Procedia PDF Downloads 185

Authors: B.L. España-Sánchez, E. Luna-Hernández, R.A. Mauricio-Sánchez, M.E. Cruz-Soto, F. Padilla-Vaca, R. Muñoz, L. Granados-López, L.R. Ovalle-Flores, J.L. Menchaca-Arredondo, G. Luna-Bárcenas

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Treatment of burn injured has been considered an important clinical problem due to the fluid control and the presence of microorganisms during the healing process. Conventional treatment includes antiseptic techniques, topical medication and surgical removal of damaged skin, to avoid bacterial growth. In order to accelerate this process, different alternatives for tissue regeneration have been explored, including artificial skin, polymers, hydrogels and hybrid materials. Some requirements consider a nonreactive organic polymer with high biocompatibility and skin adherence, avoiding bacterial infections. Chitin-derivative biopolymer such as chitosan (CS) has been used in skin regeneration following third-degree burns. The biological interest of CS is associated with the improvement of tissue cell stimulation, biocompatibility and antibacterial properties. In particular, antimicrobial properties of CS can be significantly increased when is blended with nanostructured materials. Silver-based nanocomposites have gained attention in medicine due to their high antibacterial properties against pathogens, related to their high surface area/volume ratio at nanomolar concentrations. Silver nanocomposites can be blended or synthesized with chitin-derivative biopolymers in order to obtain a biodegradable/antimicrobial hybrid with improved physic-mechanical properties. In this study, nanocomposites based on chitosan/silver nanoparticles (CS/nAg) were synthesized by the in situ chemical reduction method, improving their antibacterial properties against pathogenic bacteria and enhancing the healing process in thermal burn injuries produced in an animal model. CS/nAg was prepared in solution by the chemical reduction method, using AgNO₃ as precursor. CS was dissolved in acetic acid and mixed with different molar concentrations of AgNO₃: 0.01, 0.025, 0.05 and 0.1 M. Solutions were stirred at 95°C during 20 hours, in order to promote the nAg formation. CS/nAg solutions were placed in Petri dishes and dried, to obtain films. Structural analyses confirm the synthesis of silver nanoparticles (nAg) by means of UV-Vis and TEM, with an average size of 7.5 nm and spherical morphology. FTIR analyses showed the complex formation by the interaction of hydroxyl and amine groups with metallic nanoparticles, and surface chemical analysis (XPS) shows low concentration of Ag⁰/Ag⁺ species. Topography surface analyses by means of AFM shown that hydrated CS form a mesh with an average diameter of 10 µm. Antibacterial activity against S. aureus and P. aeruginosa was improved in all evaluated conditions, such as nAg loading and interaction time. CS/nAg nanocomposites films did not show Ag⁰/Ag⁺ release in saline buffer and rat serum after exposition during 7 days. Healing process was significantly enhanced by the presence of CS/nAg nanocomposites, inducing the production of myofibloblasts, collagen remodelation, blood vessels neoformation and epidermis regeneration after 7 days of injury treatment, by means of histological and immunohistochemistry assays. The present work suggests that hydrated CS/nAg nanocomposites can be formed a mesh, improving the bacterial penetration and the contact with embedded nAg, producing complete growth inhibition after 1.5 hours. Furthermore, CS/nAg nanocomposites improve the cell tissue regeneration in thermal burn injuries induced in rats. Synthesis of antibacterial, non-toxic, and biocompatible nanocomposites can be an important issue in tissue engineering and health care applications.

Keywords: antibacterial, chitosan, healing process, nanocomposites, silver

Procedia PDF Downloads 258

Authors: Claire Harrison, Raajit K. Rampal, Vikas Gupta, Srdan Verstovsek, Moshe Talpaz, Jean-Jacques Kiladjian, Ruben Mesa, Andrew Kuykendall, Alessandro Vannucchi, Francesca Palandri, Sebastian Grosicki, Timothy Devos, Eric Jourdan, Marielle J. Wondergem, Haifa Kathrin Al-Ali, Veronika Buxhofer-Ausch, Alberto Alvarez-Larrán, Sanjay Akhani, Rafael Muñoz-Carerras, Yury Sheykin, Gozde Colak, Morgan Harris, John Mascarenhas

Abstract:

Myelofibrosis (MF) is characterized by bone marrow fibrosis, anemia, splenomegaly and constitutional symptoms. Progressive bone marrow fibrosis results from aberrant megakaryopoeisis and expression of proinflammatory cytokines, both of which are heavily influenced by bromodomain and extraterminal domain (BET)-mediated gene regulation and lead to myeloproliferation and cytopenias. Pelabresib (CPI-0610) is an oral small-molecule investigational inhibitor of BET protein bromodomains currently being developed for the treatment of patients with MF. It is designed to downregulate BET target genes and modify nuclear factor kappa B (NF-κB) signaling. MANIFEST-2 was initiated based on data from Arm 3 of the ongoing Phase 2 MANIFEST study (NCT02158858), which is evaluating the combination of pelabresib and ruxolitinib in Janus kinase inhibitor (JAKi) treatment-naïve patients with MF. Primary endpoint analyses showed splenic and symptom responses in 68% and 56% of 84 enrolled patients, respectively. MANIFEST-2 (NCT04603495) is a global, Phase 3, randomized, double-blind, active-control study of pelabresib and ruxolitinib versus placebo and ruxolitinib in JAKi treatment-naïve patients with primary MF, post-polycythemia vera MF or post-essential thrombocythemia MF. The aim of this study is to evaluate the efficacy and safety of pelabresib in combination with ruxolitinib. Here we report updates from a recent protocol amendment. The MANIFEST-2 study schema is shown in Figure 1. Key eligibility criteria include a Dynamic International Prognostic Scoring System (DIPSS) score of Intermediate-1 or higher, platelet count ≥100 × 10^9/L, spleen volume ≥450 cc by computerized tomography or magnetic resonance imaging, ≥2 symptoms with an average score ≥3 or a Total Symptom Score (TSS) of ≥10 using the Myelofibrosis Symptom Assessment Form v4.0, peripheral blast count <5% and Eastern Cooperative Oncology Group performance status ≤2. Patient randomization will be stratified by DIPSS risk category (Intermediate-1 vs Intermediate-2 vs High), platelet count (>200 × 10^9/L vs 100–200 × 10^9/L) and spleen volume (≥1800 cm^3 vs <1800 cm^3). Double-blind treatment (pelabresib or matching placebo) will be administered once daily for 14 consecutive days, followed by a 7 day break, which is considered one cycle of treatment. Ruxolitinib will be administered twice daily for all 21 days of the cycle. The primary endpoint is SVR35 response (≥35% reduction in spleen volume from baseline) at Week 24, and the key secondary endpoint is TSS50 response (≥50% reduction in TSS from baseline) at Week 24. Other secondary endpoints include safety, pharmacokinetics, changes in bone marrow fibrosis, duration of SVR35 response, duration of TSS50 response, progression-free survival, overall survival, conversion from transfusion dependence to independence and rate of red blood cell transfusion for the first 24 weeks. Study recruitment is ongoing; 400 patients (200 per arm) from North America, Europe, Asia and Australia will be enrolled. The study opened for enrollment in November 2020. MANIFEST-2 was initiated based on data from the ongoing Phase 2 MANIFEST study with the aim of assessing the efficacy and safety of pelabresib and ruxolitinib in JAKi treatment-naïve patients with MF. MANIFEST-2 is currently open for enrollment.

Keywords: CPI-0610, JAKi treatment-naïve, MANIFEST-2, myelofibrosis, pelabresib

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