Search results for: Danilo Christen

Authors: Camille Aouinaït, Danilo Christen, Christoph Carlen

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Introduction: Small and Medium-sized Enterprises (SMEs) are facing different challenges such as pressures on environmental resources, the rise of downstream power, and trade liberalization. Remaining competitive by implementing innovations and engaging in collaborations could be a strategic solution. In Switzerland, the Federal Institute for Research in Agriculture (Agroscope), the Federal schools of technology (EPFL and ETHZ), Cantonal universities and Universities of Applied Sciences (UAS) can provide substantial inputs. UAS were developed with specific missions to match the labor markets and society needs. Research projects produce patents, publications and improved networks of scientific expertise. The study’s goal is to measure the contribution of UAS and research organization to innovation and the impact of collaborations with partners in the non-academic environment in Swiss traditional fruit production. Materials and methods: The European projects Traditional Food Network to improve the transfer of knowledge for innovation (TRAFOON) and Social Impact Assessment of Productive Interactions between science and society (SIAMPI) frame the present study. The former aims to fill the gap between the needs of traditional food producing SMEs and innovations implemented following European projects. The latter developed a method to assess the impacts of scientific research. On one side, interviews with market players have been performed to make an inventory of needs of Swiss SMEs producing apricots and berries. The participative method allowed matching the current needs and the existing innovations coming from past European projects. Swiss stakeholders (e.g. producers, retailers, an inter-branch organization of fruits and vegetables) directly rated the needs on a five-Likert scale. To transfer the knowledge to SMEs, training workshops have been organized for apricot and berries actors separately, on specific topics. On the other hand, a mapping of a social network is drawn to characterize the links between actors, with a focus on the Swiss canton of Valais and UAS Valais Wallis. Type and frequency of interactions among actors have identified thanks to interviews. Preliminary results: A list of 369 SMEs needs grouped in 22 categories was produced with 37 fulfilled questionnaires. Swiss stakeholders rated 31 needs very important. Training workshops on apricot are focusing on varietal innovations, storage, disease (bacterial blight), pest (Drosophila suzukii), sorting and rootstocks. Entrepreneurship was targeted through trademark discussions in berry production. The UAS Valais Wallis collaborated on a few projects with Agroscope along with industries, at European and national levels. Political and public bodies interfere with the central area of agricultural vulgarization that induces close relationships between the research and the practical side. Conclusions: The needs identified by Swiss stakeholders are becoming part of training workshops to incentivize innovations. The UAS Valais Wallis takes part in collaboration projects with the research environment and market players that bring innovations helping SMEs in their contextual environment. Then, a Strategic Research and Innovation Agenda will be created in order to pursue research and answer the issues facing by SMEs.

Keywords: agriculture, innovation, knowledge transfer, university and research collaboration

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Authors: Danilo Habermann, Aleksander Medella, Carla Cremon, Yusef Caceres

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This paper aims to analyze the ability of 2d point clouds features to classify different models of helicopters using radars. This method does not need to estimate the blade length, the number of blades of helicopters, and the period of their micro-Doppler signatures. It is also not necessary to generate spectrograms (or any other image based on time and frequency domain). This work transforms a radar return signal into a 2D point cloud and extracts features of it. Three classifiers are used to distinguish 9 different helicopter models in order to analyze the performance of the features used in this work. The high accuracy obtained with each of the classifiers demonstrates that the 2D point clouds features are very useful for classifying helicopters from radar signal.

Keywords: helicopter classification, point clouds features, radar, supervised classifiers

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Authors: Danilo López, Nelson Vera, Luis Pedraza

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This paper analyzes fundamental ideas and concepts related to neural networks, which provide the reader a theoretical explanation of Long Short-Term Memory (LSTM) networks operation classified as Deep Learning Systems, and to explicitly present the mathematical development of Backward Pass equations of the LSTM network model. This mathematical modeling associated with software development will provide the necessary tools to develop an intelligent system capable of predicting the behavior of licensed users in wireless cognitive radio networks.

Keywords: neural networks, multilayer perceptron, long short-term memory, recurrent neuronal network, mathematical analysis

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Authors: Danilo López, Johana Hernández, Edwin Rivas

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The use of the Multilayer Perceptron Neural Networks (MLPNN) technique is presented to estimate the future state of use of a licensed channel by primary users (PUs); this will be useful at the spectral decision stage in cognitive radio networks (CRN) to determine approximately in which time instants of future may secondary users (SUs) opportunistically use the spectral bandwidth to send data through the primary wireless network. To validate the results, sequences of occupancy data of channel were generated by simulation. The results show that the prediction percentage is greater than 60% in some of the tests carried out.

Keywords: cognitive radio, neural network, prediction, primary user

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Authors: Sandra Cuenca, Danilo Santana, Anderson Reyes

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The following project is based on the manipulation of audio signals through the Matlab software, which has an audio signal that is modified, and its resultant obtained through the auxiliary port of the computer is passed through a signal amplifier whose amplified signal is connected to a tesla coil which has a behavior like a vumeter, the flashes at the output of the tesla coil increase and decrease its intensity depending on the audio signal in the computer and also the voltage source from which it is sent. The amplified signal then passes to the tesla coil being shown in the plasma sphere with the respective flashes; this activation is given through the specified parameters that we want to give in the MATLAB algorithm that contains the digital filters for the manipulation of our audio signal sent to the tesla coil to be displayed in a plasma sphere with flashes of the combination of colors commonly pink and purple that varies according to the tone of the song.

Keywords: auxiliary port, tesla coil, vumeter, plasma sphere

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Authors: Domingo Planes Meseguer, Raffaele Danilo Esposito, Maria Del Pilar Dorado Rodriguez

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Monthly quality control checks for a Radiation Therapy Linac may be performed is a simple and efficient way once they have been standardized and protocolized. On the other hand this checks, in spite of being imperatives, require a not negligible execution times in terms of machine time and operators time. Besides it must be taken into account the amount of disposable material which may be needed together with the use of commercial software for their performing. With the aim of optimizing and standardizing mechanical-geometric checks and multi leaves collimator checks, we decided to implement a protocol which makes use of the Electronic Portal Imaging Device (EPID) available on our Linacs. The user is step by step guided by the software during the whole procedure. Acquired images are automatically analyzed by our programs all of them written using only free software.

Keywords: quality control checks, linac, radiation oncology, medical physics, free software

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Authors: Danilo López, Edwin Rivas, Fernando Pedraza

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Currently, one of the major challenges in wireless networks is the optimal use of radio spectrum, which is managed inefficiently. One of the solutions to existing problem converges in the use of Cognitive Radio (CR), as an essential parameter so that the use of the available licensed spectrum is possible (by secondary users), well above the usage values that are currently detected; thus allowing the opportunistic use of the channel in the absence of primary users (PU). This article presents the results found when estimating or predicting the future use of a spectral transmission band (from the perspective of the PU) for a chaotic type channel arrival behavior. The time series prediction method (which the PU represents) used is ANFIS (Adaptive Neuro Fuzzy Inference System). The results obtained were compared to those delivered by the RNA (Artificial Neural Network) algorithm. The results show better performance in the characterization (modeling and prediction) with the ANFIS methodology.

Keywords: ANFIS, cognitive radio, prediction primary user, RNA

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Authors: Danilo López, Edwin Rivas, Leyla López

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This paper presents the development of an algorithm that predicts the arrival of a secondary user (SU) to a base station (BS) in a cognitive network based on infrastructure, requesting a Best Effort (BE) or Real Time (RT) type of service with a determined bandwidth (BW) implementing neural networks. The algorithm dynamically uses a neural network construction technique using the geometric pyramid topology and trains a Multilayer Perceptron Neural Networks (MLPNN) based on the historical arrival of an SU to estimate future applications. This will allow efficiently managing the information in the BS, since it precedes the arrival of the SUs in the stage of selection of the best channel in CRN. As a result, the software application determines the probability of arrival at a future time point and calculates the performance metrics to measure the effectiveness of the predictions made.

Keywords: cognitive radio, base station, best effort, MLPNN, prediction, real time

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Authors: Raffaele Danilo Esposito, Domingo Planes Meseguer, Maria Del Pilar Dorado Rodriguez

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As far as we know, it is still unavailable a commercial solution which would allow to manage, openly and configurable up to user needs, the huge amount of data generated in a modern Radiation Oncology Department. Currently, available information management systems are mainly focused on Record & Verify and clinical data, and only to a small extent on physical data. Thus, results in a partial and limited use of the actually available information. In the present work we describe the implementation at our department of a centralized information management system based on a web server. Our system manages both information generated during patient planning and treatment, and information of general interest for the whole department (i.e. treatment protocols, quality assurance protocols etc.). Our objective it to be able to analyze in a simple and efficient way all the available data and thus to obtain quantitative evaluations of our treatments. This would allow us to improve our work flow and protocols. To this end we have implemented a relational data base which would allow us to use in a practical and efficient way all the available information. As always we only use license free software.

Keywords: information management system, radiation oncology, medical physics, free software

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Authors: Farid Arya, Trevor Hyde, Andrea Trevisi, Paolo Basso, Danilo Bardaro

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Heat transfer through multiple pane windows can be reduced by creating a vacuum pressure less than 0.1 Pa between the glass panes, with low emittance coatings on one or more of the internal surfaces. Fabrication of vacuum glazing (VG) requires the formation of a hermetic seal around the periphery of the glass panes together with an array of support pillars between the panes to prevent them from touching under atmospheric pressure. Atmospheric pressure and temperature differentials induce stress which can affect the integrity of the glazing. Several parameters define the stresses in VG including the glass thickness, pillar specifications, glazing dimensions and edge seal configuration. Inherent stresses in VG can result in fractures in the glass panes and failure of the edge seal. In this study, stress in VG with different glass thicknesses is theoretically studied using Finite Element Modelling (FEM). Based on the finding in this study, suggestions are made to address problems resulting from the use of thinner glass panes in the fabrication of VG. This can lead to the development of high performance, light and thin VG.

Keywords: vacuum glazing, stress, vacuum insulation, support pillars

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Authors: César Huiliñir, Danilo Villanueva, Pedro Iván Alvarez, Francisco Cubillos

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Biodrying aims at removing water from biowastes and has been mostly studied for municipal solid wastes (MSW), while few studies have dealt with secondary sludge from the paper and pulp industry. The goal of this study was to investigate the effect of initial moisture content (MC) on the batch biodrying of pulp and paper secondary sludge, using rice husks as bulking agents. Three initial MCs were studied (54, 65, and 74% w.b.) in closed batch laboratory-scale reactors under adiabatic conditions and with a constant air-flow rate (0.65 l min-1 kg-1 wet solid). The initial MC of the mixture of secondary sludge and rice husks showed a significant effect on the biodrying process. Using initial moisture content between 54-65% w.b., the solid moisture content was reduce up to 37 % w.b. in ten days, getting calorific values between 8000-9000 kJ kg-1. It was concluded that a decreasing of initial MC improves the drying rate and decreases the solid volatile consumption, therefore, the optimization of biodrying should consider this parameter.

Keywords: biodrying, secondary sludge, initial moisture content, pulp and paper industry, rice husk

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Authors: Snezana Lazic, Zarko Gacevic, Mark Holmes, Ekaterina Chernysheva, Marcus Müller, Peter Veit, Frank Bertram, Juergen Christen, Yasuhiko Arakawa, Enrique Calleja

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The study reports how the pencil-like morphology of a homoepitaxially grown GaN nanowire can be exploited for the fabrication of a thin conformal InGaN nanoshell, hosting nonpolar, semipolar and polar single photon sources (SPSs). All three SPS types exhibit narrow emission lines (FWHM~0.35 - 2 meV) and high degrees of linear optical polarization (P > 70%) in the low-temperature micro-photoluminescence (µ-PL) experiments and are characterized by a pronounced antibunching in the photon correlation measurements (gcorrected(2)(0) < 0.3). The quantum-dot-like exciton localization centers induced by compositional fluctuations within the InGaN nanoshell are identified as the driving mechanism for the single photon emission. As confirmed by the low-temperature transmission electron microscopy combined with cathodoluminescence (TEM-CL) study, the crystal region (i.e. non-polar m-, semi-polar r- and polar c-facets) hosting the single photon emitters strongly affects their emission wavelength, which ranges from ultra-violet for the non-polar to visible for the polar SPSs. The photon emission lifetime is also found to be facet-dependent and varies from sub-nanosecond time scales for the non- and semi-polar SPSs to a few nanoseconds for the polar ones. These differences are mainly attributed to facet-dependent indium content and electric field distribution across the hosting InGaN nanoshell. The hereby reported pencil-like InGaN nanoshell is the first single nanostructure able to host all three types of single photon emitters and is thus a promising building block for tunable quantum light devices integrated into future photonic and optoelectronic circuits.

Keywords: GaN nanowire, InGaN nanoshell, linear polarization, nonpolar, semipolar, polar quantum dots, single-photon sources

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Authors: Christen Kutz

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Objective: The aim of this study is to review a case series of 4 high-level Olympic and Division 1 wrestlers who experienced debilitating posterior circulation ischemic strokes during or after a competitive wrestling event and to identify risk factors, etiology and outcomes of stroke in young, healthy elite wrestlers. Background: Stroke occurs in one in 10,000 people under age 64. In young adults, the most common causes of stroke are cardiac embolism, hypercoagulable state, and vasculopathy. One-third of these strokes occur in young, fit individuals. There is little published literature about ischemic strokes that occur in wrestlers. Based on the nature of wrestling, the risk of injury or dissection to neurovascular structures may be a possible theory, but very few case reports exist. Methodology: 4 wrestlers under the age of 44 with a known history of ischemic stroke participated in individual interviews either in person or virtually. Each of the wrestlers provided their demographic information, wrestling background, clinical presentation at the time of stroke, imaging results, identification of potential risk factors, acute treatment and recovery. Results: 3 white male Division 1 wrestlers (2 Lehigh University, 1 Lock Haven University) and 1 black male 2008 Olympian experienced posterior circulation strokes. Case #1 felt a “pop” while wrestling (lateral medullary infarct, possible vertebral artery dissection); Case #2 awoke with severe vertigo, sweating, and vomiting after wrestling the previous day (left cerebellar infarct, (+) protein S deficiency); Case #3 severe vertigo, ataxia, and sensation of impending doom after wrestling earlier that week (left cerebellar infarct, hypoplastic left vertebral artery (+) anti-cardiolipin antibodies). Case #4 severe dizziness, confusion (left cerebellar stroke, vertebral artery dissection, small PFO). Conclusion: 3 wrestlers were started on anti-platelet therapy, risk factors were modified, and returned to their sport. 1 wrestler was placed on anti-coagulation and retired from competition.

Keywords: stroke, wrestling, Olympic, posterior circulation

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Authors: Hiba Dakroub, Danilo Russo, Luca Cistrone, Francesco Serra, Giovanna Fusco, Esterina De Carlo, Maria Grazia Amoroso

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The question of the origin of SARS-CoV-2 and the cycle of transmission between humans and animals is still unanswered. One serious concern associated with the SARS-CoV-2 pandemic is that the virus might spill back from humans to wildlife, which would render some animal species reservoirs of the human virus. The aim of the present study is to monitor the potential risk of SARS-CoV-2 reverse infection from humans to bats, by performing bat surveillance from different sites in Central-Southern Italy. We collected 240 droppings or saliva from 129 bats and tested them using specific and general primers of SARS-COV-2 and coronaviruses respectively. All samples, including 127 nasal swabs and 113 fecal droppings resulted negative for SARS-COV-2, and these results were confirmed by testing the samples with the Droplet Digital PCR. Also, an end-point RT-PCR was performed and no sample showed specific bands. The absence of SARS-CoV-2 in the bats we surveyed is a first step towards a better understanding of reverse transmission to bats of this virus. We hope our first contribution will encourage the establishment of systematic surveillance of wildlife, and specifically bats, to help prevent reverse zoonotic episodes that would jeopardize human health as well as biodiversity conservation and management.

Keywords: coronaviruses, bats, zoonotic viruses, spillback, SARS-CoV-2

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Authors: Ogbonna Anyanwu

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Language attrition describes the non-pathological decrease in language that had previously been acquired by an individual. It can affect some aspects of a language use or all aspects of a language use. The Igbo language, (despite its status as one of the major Nigerian languages) based on recent studies is fast losing its population of first generation speakers and therefore, increasingly becoming endangered and may be heading to extinction as warned by UNESCO if there are no conscious efforts to reverse the situation. The present paper, which contributes to the Igbo endangerment studies, examines the attrition of an aspect of the Igbo language use and practice: the indigenous Igbo wives’ pet names. It surveys the level of attrition of indigenous Igbo wives’ pet names; names which Igbo married men christen their wives upon marriage. The wives’ pet names under investigation here are specifically those which a husband traditionally christens his wife to reflect the intimate marital bond between them and also to extol his wife as an integral part of him. These pet names morphologically, are always suffixed with the compound morpheme diya which is translated as 'her husband' as in enyidiya 'her husband’s friend', obidiya 'her husband’s heart', ahudiya 'her husband’s body', ugwudiya 'her husband’s honour’, etc. The data for the study were collected through questionnaire, and oral interview from 300 male and 100 female respondents of different age groups who are married, indigenous Igbo speakers and are resident in the study areas (two Local Government Areas from two different Senatorial Zones in Abia and Imo States, south-eastern, Nigeria). Findings from the study show almost a total attrition of the Igbo indigenous wives’ pet names under study across the different age groups. For the respondents within the age group of 25-54 years, there is no more christening and bearing of the indigenous Igbo wives’ pet names by men and women respectively. This age group gives and bears pet names which the group members feel are contemporary and in line with modernity. This is a piece of evidence that the Igbo indigenous pet names’ use and practice are no longer part of the lifestyle of this group of respondents and therefore, they cannot transmit such names to their own children. For the respondents within the age group of 55-74 years, the indigenous Igbo wives’ pet names are also fading fast with less than 20% retention within the age group of 65-74 years with very few traces within the group of 55-64 years. These findings are further evidence that this aspect of Igbo language use and culture is severely threatened and may be on the verge of being lost. The loss of this aspect of the Igbo language or any aspect of the language has huge implications for the gradual and steady endangerment of the language as predicted by UNESCO.

Keywords: attrition, endangerment, practice, Igbo

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Authors: Tais Silva Lopes, Danilo Caneppele, Elizabeth Romagosa

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Surubim-do-Paraíba, Steindachneridion parahybae is a species of South American fish in critical conditions of extinction. Researches have been developed with the objective of conserving the biological material of this species. We evaluated the cooling of mature oocytes in the cryoprotective solutions containing the following alcohols: methanol, Propylene glycol and DMSO, each at concentrations of 1M, 2M and 4M, totaling nine treatments. After being submitted to treatments, the oocytes were maintained for 120 minutes in cooling to -5.52±2.58⁰C. A sample of oocytes was submitted to negative control (NC), kept in 90% L-15 solution, and positive control (PC), fertilized and taken directly to the incubator. Fertilization and hatching rates were evaluated. In order to compare the sensitivity of oocytes to embryos of the same species, the embryos maintained as CP in the previous assay were used in the free-flow stage (about 22 hours post fertilization) and submitted to the same treatments (prepared in distilled water) and also cooled for 120 min. The evaluation was done by the hatch rate. There was no fertilization rate of the oocytes submitted to the cooling with propylene glycol; the other cryoprotectants presented values of at most 3.7% of fertilization (Methanol 1M), and no treatment completed development until hatching. The cooled embryos had a significant percentage of normal larvae in all treatments, but inversely proportional to the increase in the concentration of the alcohols. DMSO 1M was the most promising treatment for embryo cooling, with 41.7% ± 20.2 of normal larvae, while mature oocytes were highly sensitive to cold.

Keywords: cryoconservation, cooling, embryos, freezing, oocytes, south American fish

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Authors: Danilo Pesce, Emilio Paolucci, Mariolina Affatato

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Looking at the future and at the post-digital era, innovations commonly tend to dismiss the old and replace it with the new. The aim of this research is to study the role that digital innovation can play alongside the information chain within the traditional sectors and the subsequent value creation opportunities that actors and stakeholders can exploit. By drawing on a wide body of literature on innovation and strategic management and by conducting a case study on the cultural heritage industry, namely Google Arts & Culture, this study shows that technology augments complements, and amplifies the way people experience their cultural interests and experience. Furthermore, the study shows a process of democratization of art since museums can exploit new digital and virtual ways to distribute art globally. Moreover, new needs arose from the 2020 pandemic that hit and forced the world to a state of cultural fasting and caused a radical transformation of the paradigm online vs. onsite. Finally, the study highlights the capabilities that are emerging at different stages of the value chain, owing to the technological innovation available in the market. In essence, this research underlines the role of Google in allowing museums to reach users worldwide, thus unlocking new mechanisms of value creation in the cultural heritage industry. Likewise, this study points out how Google provides value to users by means of increasing the provision of artworks, improving the audience engagement and virtual experience, and providing new ways to access the online contents. The paper ends with a discussion of managerial and policy-making implications.

Keywords: big data, digital platforms, digital transformation, digitization, Google Arts and Culture, stakeholders’ interests

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Authors: Danilo G. De Quadros

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Biofuels are a priority in the renewable energy agenda. The utilization of tropical grasses to ethanol production is a real opportunity to Brazil reaches the world’s leadership in biofuels production because there are 100 million hectares of sown pastures, which represent 20% of all land and 80% of agricultural areas. Basically, nowadays tropical grasses are used to raise livestock. The results obtained in this research could bring tremendous advance not only to national technology and economy but also to improve social and environmental aspects. Thus, the objective of this work was to estimate, through well-established international models, the potential of biofuels production using sown tropical pastures as feedstocks and to compare the results with sugarcane ethanol, considering state-of-art of conversion technology, advantages and limitations factors. There were used data from national and international literature about forage yield and biochemical conversion yield. Some scenarios were studied to evaluate potential advantages and limitations for cellulosic ethanol production, since non-food feedstock appeal to conversion strategies, passing through harvest, densification, logistics, environmental impacts (carbon and water cycles, nutrient recycling and biodiversity), and social aspects. If Brazil used only 1% of sown pastures to ethanol production by biochemical pathway, with average dry matter yield of 15 metric tons per hectare per year (there are results of 40 tons), resulted annually in 721 billion liters, that represents 10 times more than sugarcane ethanol projected by the Government in 2030. However, more research is necessary to take the results to commercial scale with competitive costs, considering many strategies and methods applied in ethanol production using cellulosic feedstock.

Keywords: biofuels, biochemical pathway, cellulosic ethanol, sustainability

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Authors: Danilo Di Donato

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The continuous new releases of free and open source software (FOSS) and the high costs of proprietary software -whose monopoly is characterized by closed codes and the low level of implementation and customization of software by end-users- impose a reflection on possible tools that can be chosen and adopted for the design and the representation of new steel constructions. The paper aims to show experimentation carried out to verify the actual potential and the effective applicability of FOSS supports to the BIM modeling of steel structures, particularly considering the goal of a possible workflow in order to achieve high level of development (LOD); allow effective interchange methods between different software. To this end, the examined software packages are those with open source or freeware licenses, in order to evaluate their use in architectural praxis. The test has primarily involved the experimentation of Freecad -the only Open Source software that allows a complete and integrated BIM workflow- and then the results have been compared with those of two proprietary software, Sketchup and TeklaBim Sight, which are released with a free version, but not usable for commercial purposes. The experiments carried out on Open Source, and freeware software was then compared with the outcomes that are obtained by two proprietary software, Sketchup Pro and Tekla Structure which has special modules particularly addressed to the design of steel structures. This evaluation has concerned different comparative criteria, that have been defined on the basis of categories related to the reliability, the efficiency, the potentiality, achievable LOD and user-friendliness of the analyzed software packages. In order to verify the actual outcomes of FOSS BIM for the steel structure projects, these results have been compared with a simulation related to a real case study and carried out with a proprietary software BIM modeling. Therefore, the same design theme, the project of a shelter of public space, has been developed using different software. Therefore the purpose of the contribution is to assess what are the developments and potentialities inherent in FOSS BIM, in order to estimate their effective applicability to professional practice, their limits and new fields of research they propose.

Keywords: BIM, steel buildings, FOSS, LOD

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Authors: Simone F. Medeiros, Jessica M. Fonseca, Gizelda M. Alves, Danilo M. Santos, Sérgio P. Campana-Filho, Amilton M. Santos

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Stimuli responsive and biocompatible hydrogel nanoparticles have gained special attention as systems for potential applications in controlled release of drugs to improve their therapeutic efficacy while minimizing side effects. In this work, novel solid dispersions based on thermo- and pH-responsive poly(N-vinylcaprolactam-co-itaconic acid-co-ethylene- glycol dimethacrylate) hydrogel nanoparticles embedded in chitosan matrices were prepared via spray drying for controlled release of ketoprofen. Firstly, the hydrogel nanoparticles containing ketoprofen were prepared via precipitation polymerization and their stimuli-responsive behavior, thermal properties, chemical composition, encapsulation efficiency and morphology were characterized. Then, hydrogel nanoparticles with different particles size were embedded into chitosan matrices via spray-drying. Scanning electron microscopy (SEM) analyses were performed to investigate the particles size, dispersity and morphology. Finally, ketoprofen release profiles were studied as a function of pH and temperature. Chitosan/poly(NVCL-co-IA-co-EGDMA)-ketoprofen microparticles presented spherical shape, rough surface and pronounced agglomeration, indicating that hydrogels nanoparticles loaded with ketoprofen modified the surface of chitosan matrix. The maximum encapsulation efficiency of ketoprofen into hydrogel nanoparticles was 57.8% and the electrostatic interactions between amino groups from chitosan and carboxylic groups from hydrogel nanoparticles were able to control ketoprofen release. The hydrogel nanoparticles themselves were capable to retard the release of ketoprofen-loaded until 48h of in vitro release tests, while their incorporation into chitosan matrix achieved a maximum percentage of drug release of 45%, using a mass ratio of chitosan: poly(NVCL-co-IA-co-EGDMA equal to 10:7, and 69%, using a mass ratio of chitosan: poly(NVCL-co-IA-co-EGDMA equal to 5:2.

Keywords: hydrogel nanoparticles, poly(N-vinylcaprolactam-co-itaconic acid-co-ethylene- glycol dimethacrylate), chitosan, ketoprofen, spray-drying

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Authors: Lukas J. Evans, Danilo D. Fernando

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Shrub willows (Salix spp.) have many characteristics which make them suitable for a variety of applications such as riparian zone buffers, environmental contaminant sequestration, living snow fences, and biofuel production. In some cases, these functions are limited due to physical or financial obstacles associated with the number of individuals needed to reasonably satisfy that purpose. One way to increase the efficiency of willows is to bioengineer them with the genetic improvements suitable for the desired use. To accomplish this goal, an optimized in vitro transformation protocol via Agrobacterium tumefaciens is necessary to reliably express genes of interest. Therefore, the aim of this study is to observe the influence of tissue culture with different willow cultivars, hormones, and explants on the percentage of calli expressing reporter gene green florescent protein (GFP) to find ideal transformation conditions. Each callus was produced from 1 month old open-pollinated seedlings of three Salix miyabeana cultivars (‘SX61’, ‘WT1’, and ‘WT2’) from three different explants (lamina, petiole, and internodes). Explants were cultured for 1 month on an MS media with different concentrations of 6-Benzylaminopurine (BAP) and 1-Naphthaleneacetic acid (NAA) (No hormones, 1 mg⁻¹L BAP only, 3 mg⁻¹L NAA only, 1 mg⁻¹L BAP and 3 mg⁻¹L NAA, and 3 mg⁻¹L BAP and 1 mg⁻¹L NAA) to produce a callus. Samples were then treated with Agrobacterium tumefaciens at an OD600 of 0.6-0.8 to insert the transgene GFP for 30 minutes, co-cultivated for 72 hours, and selected on the same media type they were cultured on with added 7.5 mg⁻¹L of Hygromycin for 1 week before GFP visualization under a UV dissecting scope. Percentage of GFP expressing calli as well as the average number of fluorescing GFP units per callus were recorded and results were evaluated through an ANOVA test (α = 0.05). The WT1 internode-derived calli on media with 3 mg-1L NAA+1 mg⁻¹L BAP and mg⁻¹L BAP alone produced a significantly higher percentage of GFP expressing calli than each other group (19.1% and 19.4%, respectively). Additionally, The WT1 internode group cultured with 3 mg⁻¹L NAA+1 mg⁻¹L BAP produced an average of 2.89 GFP units per callus while the group cultivated with 1 mg⁻¹L BAP produced an average of 0.84 GFP units per callus. In conclusion, genotype, explant choice, and hormones all play a significant role in increasing successful transformation in willows. Future studies to produce whole callus GFP expression and subsequent plantlet regeneration are necessary for a complete willow transformation protocol.

Keywords: agrobacterium, callus, Salix, tissue culture

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Authors: Danilo A. F. Fontes, Magaly A. M.Lyra, Maria L. C. Moura, Leslie R. M. Ferraz, Salvana P. M. Costa, Amanda C. Q. M. Vieira, Larissa A. Rolim, Giovanna C. R. M. Schver, Ping I. Lee, Severino Alves-Júnior, José L. Soares-Sobrinho, Pedro J. Rolim-Neto

Abstract:

Efavirenz (EFV) is a first-choice drug in antiretroviral therapy with high efficacy in the treatment of infection by Human Immunodeficiency Virus, which causes Acquired Immune Deficiency Syndrome (AIDS). EFV has low solubility in water resulting in a decrease in the dissolution rate and, consequently, in its bioavailability. Among the technological alternatives to increase solubility, the Lamellar Double Hydroxides (LDH) have been applied in the development of systems with poorly water-soluble drugs. The use of analytical techniques such as X-Ray Diffraction (XRD), Infrared Spectroscopy (IR) and Differential Scanning Calorimetry (DSC) allowed the elucidation of drug interaction with the lamellar compounds. The objective of this work was to characterize and develop the binary systems with EFV and LDH in order to increase the solubility of the drug. The LDH-CaAl was synthesized by the method of co-precipitation from salt solutions of calcium nitrate and aluminum nitrate in basic medium. The systems EFV-LDH and their physical mixtures (PM) were obtained at different concentrations (5-60% of EFV) using the solvent technique described by Takahashi & Yamaguchi (1991). The characterization of the systems and the PM’s was performed by XRD techniques, IR, DSC and dissolution test under non-sink conditions. The results showed improvements in the solubility of EFV when associated with LDH, due to a possible change in its crystal structure and formation of an amorphous material. From the DSC results, one could see that the endothermic peak at 173°C, temperature that correspond to the melting process of EFZ in the crystal form, was present in the PM results. For the EFZ-LDH systems (with 5, 10 and 30% of drug loading), this peak was not observed. XRD profiles of the PM showed well-defined peaks for EFV. Analyzing the XRD patterns of the systems, it was found that the XRD profiles of all the systems showed complete attenuation of the characteristic peaks of the crystalline form of EFZ. The IR technique showed that, in the results of the PM, there was the appearance of one band and overlap of other bands, while the IR results of the systems with 5, 10 and 30% drug loading showed the disappearance of bands and a few others with reduced intensity. The dissolution test under non-sink conditions showed that systems with 5, 10 and 30% drug loading promoted a great increase in the solubility of EFV, but the system with 10% of drug loading was the only one that could keep substantial amount of drug in solution at different pHs.

Keywords: Efavirenz, Lamellar Double Hydroxides, Pharmaceutical Techonology, Solubility

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Authors: Salvana P. M. Costa, Tarcyla A. Gomes, Giovanna C. R. M. Schver, Leslie R. M. Ferraz, Cristovão R. Silva, Magaly A. M. Lyra, Danilo A. F. Fonte, Larissa A. Rolim, Amanda C. Q. M. Vieira, Miracy M. Albuquerque, Pedro J. Rolim-neto

Abstract:

Efavirenz (EFV) is considered one of the most widely used anti-HIV drugs. However, it is classified as a drug class II (poorly soluble, highly permeable) according to the biopharmaceutical classification system, presenting problems of absorption in the gastrointestinal tract and thereby inadequate bioavailability for its therapeutic action. This study aimed to overcome these barriers by developing and obtaining solid dispersions (SD) in order to increase the EFZ bioavailability. For the development of SD with EFV, theoretical and practical studies were initially performed. Thus, there was a choice of a carrier to be used. For this, it was analyzed the various criteria such as glass transition temperature of the polymer, intra- and intermolecular interactions of hydrogen bonds between drug and polymer, the miscibility between the polymer and EFV. The choice of the obtainment method of the SD came from the analysis of which method is the most consolidated in both industry and literature. Subsequently, the choice of drug and carrier concentrations in the dispersions was carried out. In order to obtain DS to present the drug in its amorphous form, as the DS were obtained, they were analyzed by X-ray diffraction (XRD). SD are more stable the higher the amount of polymer present in the formulation. With this assumption, a SD containing 10% of drug was initially prepared and then this proportion was increased until the XRD showed the presence of EFV in its crystalline form. From this point, it was not produced SD with a higher concentration of drug. Thus, it was allowed to select PVP-K30, PVPVA 64 and the SOLUPLUS formulation as carriers, once it was possible the formation of hydrogen bond between EFV and polymers since these have hydrogen acceptor groups capable of interacting with the donor group of the drug hydrogen. It is worth mentioning also that the films obtained, independent of concentration used, were presented homogeneous and transparent. Thus, it can be said that the EFV is miscible in the three polymers used in the study. The SD and Physical Mixtures (PM) with these polymers were prepared by the solvent method. The EFV diffraction profile showed main peaks at around 2θ of 6,24°, in addition to other minor peaks at 14,34°, 17,08°, 20,3°, 21,36° and 25,06°, evidencing its crystalline character. Furthermore, the polymers showed amorphous nature, as evidenced by the absence of peaks in their XRD patterns. The XRD patterns showed the PM overlapping profile of the drug with the polymer, indicating the presence of EFV in its crystalline form. Regardless the proportion of drug used in SD, all the samples showed the same characteristics with no diffraction peaks EFV, demonstrating the behavior amorphous products. Thus, the polymers enabled, effectively, the formation of amorphous SD, probably due to the potential hydrogen bonds between them and the drug. Moreover, the XRD analysis showed that the polymers were able to maintain its amorphous form in a concentration of up to 80% drug.

Keywords: amorphous form, Efavirenz, solid dispersions, solubility

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Authors: Danilo F. Rodrigues, Hérida Regina N. Salgado

Abstract:

Introduction: The emergence of resistant microorganisms to a large number of clinically approved antimicrobials has been increasing, which restrict the options for the treatment of bacterial infections. As a strategy, drugs with high antimicrobial activities are in evidence. Stands out a class of antimicrobial, the cephalosporins, having as fourth generation cefepime (CEF) a semi-synthetic product which has activity against various Gram-positive bacteria (e.g. oxacillin resistant Staphylococcus aureus) and Gram-negative (e.g. Pseudomonas aeruginosa) aerobic. There are few studies in the literature regarding the development of microbiological methodologies for the analysis of this antimicrobial, so researches in this area are highly relevant to optimize the analysis of this drug in the industry and ensure the quality of the marketed product. The development of microbiological methods for the analysis of antimicrobials has gained strength in recent years and has been highlighted in relation to physicochemical methods, especially because they make possible to determine the bioactivity of the drug against a microorganism. In this context, the aim of this work was the development and validation of a microbiological method for quantitative analysis of CEF in powder lyophilized for injectable solution by turbidimetric assay. Method: For performing the method, Staphylococcus aureus ATCC 6538 IAL 2082 was used as the test microorganism and the culture medium chosen was the Casoy broth. The test was performed using temperature control (35.0 °C ± 2.0 °C) and incubated for 4 hours in shaker. The readings of the results were made at a wavelength of 530 nm through a spectrophotometer. The turbidimetric microbiological method was validated by determining the following parameters: linearity, precision (repeatability and intermediate precision), accuracy and robustness, according to ICH guidelines. Results and discussion: Among the parameters evaluated for method validation, the linearity showed results suitable for both statistical analyses as the correlation coefficients (r) that went 0.9990 for CEF reference standard and 0.9997 for CEF sample. The precision presented the following values 1.86% (intraday), 0.84% (interday) and 0.71% (between analyst). The accuracy of the method has been proven through the recovery test where the mean value obtained was 99.92%. The robustness was verified by the parameters changing volume of culture medium, brand of culture medium, incubation time in shaker and wavelength. The potency of CEF present in the samples of lyophilized powder for injectable solution was 102.46%. Conclusion: The turbidimetric microbiological method proposed for quantification of CEF in lyophilized powder for solution for injectable showed being fast, linear, precise, accurate and robust, being in accordance with all the requirements, which can be used in routine analysis of quality control in the pharmaceutical industry as an option for microbiological analysis.

Keywords: cefepime hydrochloride, quality control, turbidimetric assay, validation

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Authors: Pedro J. Rolim-Neto, Leslie R. M. Ferraz, Fabiana L. A. Santos, Pablo A. Ferreira, Ricardo T. L. Maia-Jr., Magaly A. M. Lyra, Danilo A F. Fonte, Salvana P. M. Costa, Amanda C. Q. M. Vieira, Larissa A. Rolim

Abstract:

Initially developed as an antimalarial agent, hydroxychloroquine (HCQ) sulfate is often used as a slow-acting antirheumatic drug in the treatment of disorders of connective tissue. The United States Pharmacopeia (USP) 37 provides a reversed-phase HPLC method for quantification of HCQ. However, this method was not reproducible, producing asymmetric peaks in a long analysis time. The asymmetry of the peak may cause an incorrect calculation of the concentration of the sample. Furthermore, the analysis time is unacceptable, especially regarding the routine of a pharmaceutical industry. The aiming of this study was to develop a fast, easy and efficient method for quantification of HCQ sulfate by High Performance Liquid Chromatography (HPLC) based on the Quality by Design (QbD) methodology. This method was optimized in terms of peak symmetry using the surface area graphic as the Design of Experiments (DoE) and the tailing factor (TF) as an indicator to the Design Space (DS). The reference method used was that described at USP 37 to the quantification of the drug. For the optimized method, was proposed a 33 factorial design, based on the QbD concepts. The DS was created with the TF (in a range between 0.98 and 1.2) in order to demonstrate the ideal analytical conditions. Changes were made in the composition of the USP mobile-phase (USP-MP): USP-MP: Methanol (90:10 v/v, 80:20 v/v and 70:30 v/v), in the flow (0.8, 1.0 and 1.2 mL) and in the oven temperature (30, 35, and 40ºC). The USP method allowed the quantification of drug in a long time (40-50 minutes). In addition, the method uses a high flow rate (1,5 mL.min-1) which increases the consumption of expensive solvents HPLC grade. The main problem observed was the TF value (1,8) that would be accepted if the drug was not a racemic mixture, since the co-elution of the isomers can become an unreliable peak integration. Therefore, the optimization was suggested in order to reduce the analysis time, aiming a better peak resolution and TF. For the optimization method, by the analysis of the surface-response plot it was possible to confirm the ideal setting analytical condition: 45 °C, 0,8 mL.min-1 and 80:20 USP-MP: Methanol. The optimized HPLC method enabled the quantification of HCQ sulfate, with a peak of high resolution, showing a TF value of 1,17. This promotes good co-elution of isomers of the HCQ, ensuring an accurate quantification of the raw material as racemic mixture. This method also proved to be 18 times faster, approximately, compared to the reference method, using a lower flow rate, reducing even more the consumption of the solvents and, consequently, the analysis cost. Thus, an analytical method for the quantification of HCQ sulfate was optimized using QbD methodology. This method proved to be faster and more efficient than the USP method, regarding the retention time and, especially, the peak resolution. The higher resolution in the chromatogram peaks supports the implementation of the method for quantification of the drug as racemic mixture, not requiring the separation of isomers.

Keywords: analytical method, hydroxychloroquine sulfate, quality by design, surface area graphic

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Authors: Guilherme M. Pereira, Kimmo Teinïla, Danilo Custódio, Risto Hillamo, Célia Alves, Pérola de C. Vasconcellos

Abstract:

South American large cities often present serious air pollution problems and their atmosphere composition is influenced by a variety of emissions sources. The South American Emissions Megacities, and Climate project (SAEMC) has focused on the study of emissions and its influence on climate in the South American largest cities and it also included Lima (Peru) and Medellin (Colombia), sites where few studies of the genre were done. Lima is a coastal city with more than 8 million inhabitants and the second largest city in South America. Medellin is a 2.5 million inhabitants city and second largest city in Colombia; it is situated in a valley. The samples were collected in quartz fiber filters in high volume samplers (Hi-Vol), in 24 hours of sampling. The samples were collected in intensive campaigns in both sites, in July, 2010. Several species were determined in the aerosol samples of Lima and Medellin. Organic and elemental carbon (OC and EC) in thermal-optical analysis; biomass burning tracers (levoglucosan - Lev, mannosan - Man and galactosan - Gal) in high-performance anion exchange ion chromatography with mass spectrometer detection; water soluble ions in ion chromatography. The average particulate matter was similar for both campaigns, the PM10 concentrations were above the recommended by World Health Organization (50 µg m⁻³ – daily limit) in 40% of the samples in Medellin, while in Lima it was above that value in 15% of the samples. The average total ions concentration was higher in Lima (17450 ng m⁻³ in Lima and 3816 ng m⁻³ in Medellin) and the average concentrations of sodium and chloride were higher in this site, these species also had better correlations (Pearson’s coefficient = 0,63); suggesting a higher influence of marine aerosol in the site due its location in the coast. Sulphate concentrations were also much higher at Lima site; which may be explained by a higher influence of marine originated sulphate. However, the OC, EC and monosaccharides average concentrations were higher at Medellin site; this may be due to the lower dispersion of pollutants due to the site’s location and a larger influence of biomass burning sources. The levoglucosan average concentration was 95 ng m⁻³ for Medellin and 16 ng m⁻³ and OC was well correlated with levoglucosan (Pearson’s coefficient = 0,86) in Medellin; suggesting a higher influence of biomass burning over the organic aerosol in this site. The Lev/Man ratio is often related to the type of biomass burned and was close to 18, similar to the observed in previous studies done at biomass burning impacted sites in the Amazon region; backward trajectories also suggested the transport of aerosol from that region. Biomass burning appears to have a larger influence on the air quality in Medellin, in addition the vehicular emissions; while Lima showed a larger influence of marine aerosol during the study period.

Keywords: aerosol transport, atmospheric particulate matter, biomass burning, SAEMC project

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Authors: Leslie Raphael de M. Ferraz, Salvana Priscylla M. Costa, Tarcyla de A. Gomes, Giovanna Christinne R. M. Schver, Cristóvão R. da Silva, Magaly Andreza M. de Lyra, Danilo Augusto F. Fontes, Larissa A. Rolim, Amanda Carla Q. M. Vieira, Miracy M. de Albuquerque, Pedro J. Rolim-Neto

Abstract:

Efavirenz (EFV) is a drug used as first-line treatment of AIDS. However, it has poor aqueous solubility and wettability, presenting problems in the gastrointestinal tract absorption and bioavailability. One of the most promising strategies to improve the solubility is the use of solid dispersions (SD). Therefore, this study aimed to characterize SD EFZ with the polymers: PVP-K30, PVPVA 64 and SOLUPLUS in order to find an optimal formulation to compose a future pharmaceutical product for AIDS therapy. Initially, Physical Mixtures (PM) and SD with the polymers were obtained containing 10, 20, 50 and 80% of drug (w/w) by the solvent method. The best formulation obtained between the SD was selected by in vitro dissolution test. Finally, the drug-carrier system chosen, in all ratios obtained, were analyzed by the following techniques: Differential Scanning Calorimetry (DSC), polarization microscopy, Scanning Electron Microscopy (SEM) and spectrophotometry of absorption in the region of infrared (IR). From the dissolution profiles of EFV, PM and SD, the values of area Under The Curve (AUC) were calculated. The data showed that the AUC of all PM is greater than the isolated EFV, this result is derived from the hydrophilic properties of the polymers thus favoring a decrease in surface tension between the drug and the dissolution medium. In adittion, this ensures an increasing of wettability of the drug. In parallel, it was found that SD whom had higher AUC values, were those who have the greatest amount of polymer (with only 10% drug). As the amount of drug increases, it was noticed that these results either decrease or are statistically similar. The AUC values of the SD using the three different polymers, followed this decreasing order: SD PVPVA 64-EFV 10% > SD PVP-K30-EFV 10% > SD Soluplus®-EFV 10%. The DSC curves of SD’s did not show the characteristic endothermic event of drug melt process, suggesting that the EFV was converted to its amorphous state. The analysis of polarized light microscopy showed significant birefringence of the PM’s, but this was not observed in films of SD’s, thus suggesting the conversion of the drug from the crystalline to the amorphous state. In electron micrographs of all PM, independently of the percentage of the drug, the crystal structure of EFV was clearly detectable. Moreover, electron micrographs of the SD with the two polymers in different ratios investigated, we observed the presence of particles with irregular size and morphology, also occurring an extensive change in the appearance of the polymer, not being possible to differentiate the two components. IR spectra of PM corresponds to the overlapping of polymer and EFV bands indicating thereby that there is no interaction between them, unlike the spectra of all SD that showed complete disappearance of the band related to the axial deformation of the NH group of EFV. Therefore, this study was able to obtain a suitable formulation to overcome the solubility limitations of the EFV, since SD PVPVA 64-EFZ 10% was chosen as the best system in delay crystallization of the prototype, reaching higher levels of super saturation.

Keywords: characterization, dissolution, Efavirenz, solid dispersions

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