Search results for: Carmen Flores

Authors: Mylena McCoggle, Shyra Wilson, Andrea Rivera, Rocio Alba-Flores

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This work introduces the use of EMGs (electromyography) from muscle sensors to develop an Artificial Neural Network (ANN) for pattern recognition to control a small unmanned aerial vehicle. The objective of this endeavor exhibits interfacing drone applications beyond manual control directly. MyoWare Muscle sensor contains three EMG electrodes (dual and single type) used to collect signals from the posterior (extensor) and anterior (flexor) forearm and the bicep. Collection of raw voltages from each sensor were connected to an Arduino Uno and a data processing algorithm was developed with the purpose of interpreting the voltage signals given when performing flexing, resting, and motion of the arm. Each sensor collected eight values over a two-second period for the duration of one minute, per assessment. During each two-second interval, the movements were alternating between a resting reference class and an active motion class, resulting in controlling the motion of the drone with left and right movements. This paper further investigated adding up to three sensors to differentiate between hand gestures to control the principal motions of the drone (left, right, up, and land). The hand gestures chosen to execute these movements were: a resting position, a thumbs up, a hand swipe right motion, and a flexing position. The MATLAB software was utilized to collect, process, and analyze the signals from the sensors. The protocol (machine learning tool) was used to classify the hand gestures. To generate the input vector to the ANN, the mean, root means squared, and standard deviation was processed for every two-second interval of the hand gestures. The neuromuscular information was then trained using an artificial neural network with one hidden layer of 10 neurons to categorize the four targets, one for each hand gesture. Once the machine learning training was completed, the resulting network interpreted the processed inputs and returned the probabilities of each class. Based on the resultant probability of the application process, once an output was greater or equal to 80% of matching a specific target class, the drone would perform the motion expected. Afterward, each movement was sent from the computer to the drone through a Wi-Fi network connection. These procedures have been successfully tested and integrated into trial flights, where the drone has responded successfully in real-time to predefined command inputs with the machine learning algorithm through the MyoWare sensor interface. The full paper will describe in detail the database of the hand gestures, the details of the ANN architecture, and confusion matrices results.

Keywords: artificial neural network, biosensors, electromyography, machine learning, MyoWare muscle sensors, Arduino

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Authors: Henry Gonzalo Acurio Flores, Alvaro Nicolas Corral Naveda, Juan Francisco Fonseca Palacios

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In Ecuador, subventions on fossil fuels for the road transportation sector have always been part of its economy throughout time, mainly because of demagogy and populism from political leaders. It is clearly seen that the government cannot maintain the subsidies anymore due to its commercial balance and its general state budget; subsidies are a key barrier to implementing the use of cleaner technologies. However, during the last few months, the elimination of subsidies has been done gradually with the purpose of reaching international prices. It is expected that with this measure, the population will opt for other means of transportation, and in a certain way, it will promote the use of private electric vehicles and public, e.g., taxis and buses (urban transport). Considering the three main elements of sustainable development, an analysis of the social, economic, and environmental impacts of eliminating subsidies will be generated at the country level. To achieve this, four scenarios will be developed in order to determine how the subsidies will contribute to the promotion of electro-mobility. 1) A Business as Usual BAU scenario; 2) the introduction of 10 000 electric vehicles by 2025; 3) the introduction of 100 000 electric vehicles by 2030; 4) the introduction of 750 000 electric vehicles by 2040 (for all the scenarios buses, taxis, lightweight duty vehicles, and private vehicles will be introduced, as it is established in the National Electro Mobility Strategy for Ecuador). The Low Emissions Analysis Platform (LEAP) will be used, and it will be suitable to determine the cost for the government in terms of importing derivatives for fossil fuels and the cost of electricity to power the electric fleet that can be changed. The elimination of subventions generates fiscal resources for the state that can be used to develop other kinds of projects that will benefit Ecuadorian society. It will definitely change the energy matrix, and it will provide energy security for the country; it will be an opportunity for the government to incentivize a greater introduction of renewable energies, e.g., solar, wind, and geothermal. At the same time, it will also reduce greenhouse gas emissions (GHG) from the transportation sector, considering its mitigation potential, which as a result, will ameliorate the inhabitant quality of life by improving the quality of air, therefore reducing respiratory diseases associated with exhaust emissions, consequently, achieving sustainability, the Sustainable Development Goals (SDGs), and complying with the agreements established in the Paris Agreement COP 21 in 2015. Electro mobility in Latin America and the Caribbean can only be achieved by the implementation of the right policies at the central government, which need to be accompanied by a National Urban Mobility Policy (NUMP) and can encompass a greater vision to develop holistic, sustainable transport systems at local governments.

Keywords: electro mobility, energy, policy, sustainable transportation

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Authors: Domingos Viegas, Miguel Almeida, Carmen Rocha, Ilda Novo, Yolanda Luna

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Forest fires along the more than 1200km of the Spanish-Portuguese border are more and more frequent, currently achieving around 2000 fire events per year. Some of these events develop to large international wildfire requiring concerted operations based on shared information between the two countries. The fire event of Valencia de Alcantara (2003) causing several fatalities and more than 13000ha burnt, is a reference example of these international events. Currently, Portugal and Spain have a specific cross-border cooperation protocol on wildfires response for a strip of about 30km (15 km for each side). It is recognized by public authorities the successfulness of this collaboration however it is also assumed that this cooperation should include more functionalities such as the development of a common risk information system for transboundary fire events. Since Portuguese and Spanish authorities use different approaches to determine the fire risk indexes inputs and different methodologies to assess the fire risk, sometimes the conjoint firefighting operations are jeopardized since the information is not harmonized and the understanding of the situation by the civil protection agents from both countries is not unique. Thus, a methodology aiming the harmonization of the fire risk calculation and perception by Portuguese and Spanish Civil protection authorities is hereby presented. The final results are presented as well. The fire risk index used in this work is the Canadian Fire Weather Index (FWI), which is based on meteorological data. The FWI is limited on its application as it does not take into account other important factors with great effect on the fire appearance and development. The combination of these factors is very complex since, besides the meteorology, it addresses several parameters of different topics, namely: sociology, topography, vegetation and soil cover. Therefore, the meaning of FWI values is different from region to region, according the specific characteristics of each region. In this work, a methodology for FWI calibration based on the number of fire occurrences and on the burnt area in the transboundary regions of Portugal and Spain, in order to assess the fire risk based on calibrated FWI values, is proposed. As previously mentioned, the cooperative firefighting operations require a common perception of the information shared. Therefore, a common classification of the fire risk for the fire events occurred in the transboundary strip is proposed with the objective of harmonizing this type of information. This work is integrated in the ECHO project SpitFire - Spanish-Portuguese Meteorological Information System for Transboundary Operations in Forest Fires, which aims the development of a web platform for the sharing of information and supporting decision tools to be used in international fire events involving Portugal and Spain.

Keywords: data harmonization, FWI, international collaboration, transboundary wildfires

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Authors: Florence Roselle Adalin, Misha Louise Delariarte, Fabbette Laire Lagas, Sarah Emanuelle Mejia, Lika Mizukoshi, Irish Paullen Palomeno, Gibrianne Alistaire Ramos, Danica Pauline Ramos, Josefina Tuazon, Jo Leah Flores

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Pain, often a missed and undertreated symptom, affects the quality of life of individuals. Nurses are key players in providing effective pain management to decrease morbidity and mortality of patients in pain. Nurses’ knowledge and attitude on pain greatly affect their ability on assessment and management. The Pain Society of the Philippines recognized the inadequacy and inaccessibility of data on the knowledge, skills, and attitude of nurses on pain management in the country. This study may be the first of its kind in the county, giving it the potential to contribute greatly to nursing education and practice through providing valuable baseline data. Objectives: This study aims to describe the level of knowledge and attitude, and current practices of nurses on pain assessment and management; and determine the relationship of nurses’ knowledge and attitude with years of experience, training on pain management and clinical area of practice. Methodology: A survey research design was employed. Four hospitals were selected through purposive sampling. A total of 235 Medical-Surgical Unit and Intensive Care Unit (ICU) nurses participated in the study. The tool used is a combination of demographic survey, Nurses’ Knowledge and Attitude Survey Regarding Pain (NKASRP), Acute Pain Evidence Based Practice Questionnaire (APEBPQ) with self-report questions on non-pharmacologic pain management. The data obtained was analysed using descriptive statistics, two sample T-tests for clinical areas and training; and Pearson product correlation to identify relationship of level of knowledge and attitude with years of experience. Results and Analysis: The mean knowledge and attitude score of the nurses was 47.14%. Majority answered ‘most of the time’ or ‘all the time’ on 84.12% of practice items on pain assessment, implementation of non-pharmacologic interventions, evaluation and documentation. Three of 19 practice items describing morphine and opioid administration in special populations were only done ‘a little of the time’. Most utilized non-pharmacologic interventions were deep breathing exercises (79.66%), massage therapy (27.54%), and ice therapy (26.69%). There was no significant relationship between knowledge scores and years of clinical experience (p = 0.05, r= -0.09). Moreover, there was not enough evidence to show difference in nurses’ knowledge and attitude scores in relation to presence of training (p = 0.41) or areas (Medical-Surgical or ICU) of clinical practice (p = 0.53). Conclusion and Recommendations: Findings of the study showed that the level of knowledge and attitude of nurses on pain assessment and management is suboptimal; and no relationship between nurses’ knowledge and attitude and years of experience. It is recommended that further studies look into the nursing curriculum on pain education, culture-specific pain management protocols and evidence-based practices in the country.

Keywords: knowledge and attitude, nurses, pain management, practices on pain management

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Authors: Carmen Noheda

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This text provides a reflection on the way of thinking about the study of the history of music by examining the production of historiography in Spain at the turn of the century. Based on concepts developed by the historical theorist Jörn Rüsen, the article focuses on the following aspects: the theoretical artifacts that structure the interpretation of the limits of writing the history of music, the narrative patterns used to give meaning to the discourse of history, and the orientation context that functions as a source of criteria of significance for both interpretation and representation. This analysis intends to show that historical music theory is not only a means to abstractly explore the complex questions connected to the production of historical knowledge, but also a tool for obtaining concrete images about the intellectual practice of professional musicologists. Writing about the historiography of contemporary Spanish music is a task that requires both a knowledge of the history that is being written and investigated, as well as a familiarity with current theoretical trends and methodologies that allow for the recognition and definition of the different tendencies that have arisen in recent decades. With the objective of carrying out these premises, this project takes as its point of departure the 'immediate historiography' in relation to Spanish music at the beginning of the 21st century. The hesitation that Spanish musicology has shown in opening itself to new anthropological and sociological approaches, along with its rigidity in the face of the multiple shifts in dynamic forms of thinking about history, have produced a standstill whose consequences can be seen in the delayed reception of the historiographical revolutions that have emerged in the last century. Methodologically, this essay is underpinned by Rüsen’s notion of the disciplinary matrix, which is an important contribution to the understanding of historiography. Combined with his parallel conception of differing paradigms of historiography, it is useful for analyzing the present-day forms of thinking about the history of music. Following these theories, the article will in the first place address the characteristics and identification of present historiographical currents in Spanish musicology to thereby carry out an analysis based on the theories of Rüsen. Finally, it will establish some considerations for the future of musical historiography, whose atrophy has not only fostered the maintenance of an ingrained positivist tradition, but has also implied, in the case of Spain, an absence of methodological schools and an insufficient participation in international theoretical debates. An update of fundamental concepts has become necessary in order to understand that thinking historically about music demands that we remember that subjects are always linked by reciprocal interdependencies that structure and define what it is possible to create. In this sense, the fundamental aim of this research departs from the recognition that the history of music is embedded in the conditions that make it conceivable, communicable and comprehensible within a society.

Keywords: historiography, Jörn Rüssen, Spanish musicology, theory of history of music

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Authors: Itzel Amaro-Estrada, Eduardo Vergara-Rivera, Virginia Juarez-Flores, Mayra Cobaxin-Cardenas, Rosa Estela Quiroz, Jesus F. Preciado, Sergio Rodriguez-Camarillo

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Bovine anaplasmosis is an infectious, tick-borne disease caused mainly by Anaplasma marginale; typical signs include anemia, fever, abortion, weight loss, decreased milk production, jaundice, and potentially death. Sick bovine can recover when antibiotics are administered; however, it usually remains as carrier for life, being a risk of infection for susceptible cattle. Anaplasma marginale is an obligate intracellular Gram-negative bacterium with genetic composition highly diverse among geographical isolates. There are currently no vaccines fully effective against bovine anaplasmosis; therefore, the economic losses due to disease are present. Vaccine formulation became a hard task for several pathogens as Anaplasma marginale, but peptide-based vaccines are an interesting proposal way to induce specific responses. Phage-displayed peptide libraries have been proved one of the most powerful technologies for identifying specific ligands. Screening of these peptides libraries is also a tool for studying interactions between proteins or peptides. Thus, it has allowed the identification of ligands recognized by polyclonal antiserums, and it has been successful for the identification of relevant epitopes in chronic diseases and toxicological conditions. Protective immune response to bovine anaplasmosis includes high levels of immunoglobulins subclass G2 (IgG2) but not subclass IgG1. Therefore, IgG2 from the serum of protected bovine can be useful to identify ligands, which can be part of an immunogen for cattle. In this work, phage display random peptide library Ph.D. ™ -12 was incubating with IgG2 or blood sera of immunized bovines against A. marginale as targets. After three rounds of biopanning, several candidates were selected for additional analysis. Subsequently, their reactivity with sera immunized against A. marginale, as well as with positive and negative sera to A. marginale was evaluated by immunoassays. A collection of recognized peptides tested by ELISA was generated. More than three hundred phage-peptides were separately evaluated against molecules which were used during panning. At least ten different peptides sequences were determined from their nucleotide composition. In this approach, three phage-peptides were selected by their binding and affinity properties. In the case of the development of vaccines or diagnostic reagents, it is important to evaluate the immunogenic and antigenic properties of the peptides. Immunogenic in vitro and in vivo behavior of peptides will be assayed as synthetic and as phage-peptide for to determinate their vaccine potential. Acknowledgment: This work was supported by grant SEP-CONACYT 252577 given to I. Amaro-Estrada.

Keywords: bovine anaplasmosis, peptides, phage display, veterinary vaccines

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Authors: Cristina Costescu, Carmen David, Adrian Roșan

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Executive functions (EF) and emotion regulation strategies are processes that allow individuals to function in an adaptative way and to be goal-oriented, which is essential for success in daily living activities, at school, or in social contexts. The Emotion Regulation and Executive Functioning Scale for Children and Adolescents (REMEX) represents an empirically based tool (based on the model of EF developed by Diamond) for evaluating significant dimensions of child and adolescent EFs and emotion regulation strategies, mainly in school contexts. The instrument measures the following dimensions: working memory, inhibition, cognitive flexibility, executive attention, planning, emotional control, and emotion regulation strategies. Building the instrument involved not only a top-down process, as we selected the content in accordance with prominent models of FE, but also a bottom-up one, as we were able to identify valid contexts in which FE and ER are put to use. For the construction of the instrument, we implemented three focus groups with teachers and other professionals since the aim was to develop an accurate, objective, and ecological instrument. We used the focus group method in order to address each dimension and to yield a bank of items to be further tested. Each dimension is addressed through a task that the examiner will apply and through several items derived from the main task. For the validation of the instrument, we plan to use item response theory (IRT), also known as the latent response theory, that attempts to explain the relationship between latent traits (unobservable cognitive processes) and their manifestations (i.e., observed outcomes, responses, or performance). REMEX represents an ecological scale that integrates a current scientific understanding of emotion regulation and EF and is directly applicable to school contexts, and it can be very useful for developing intervention protocols. We plan to test his convergent validity with the Childhood Executive Functioning Inventory (CHEXI) and Emotion Dysregulation Inventory (EDI) and divergent validity between a group of typically developing children and children with neurodevelopmental disorders, aged between 6 and 9 years old. In a previous pilot study, we enrolled a sample of 40 children with autism spectrum disorders and attention-deficit/hyperactivity disorder aged 6 to 12 years old, and we applied the above-mentioned scales (CHEXI and EDI). Our results showed that deficits in planning, bebavior regulation, inhibition, and working memory predict high levels of emotional reactivity, leading to emotional and behavioural problems. Considering previous results, we expect our findings to provide support for the validity and reliability of the REMEX version as an ecological instrument for assessing emotion regulation and EF in children and for key features of its uses in intervention protocols.

Keywords: executive functions, emotion regulation, children, item response theory, focus group

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Authors: Leticia Lengler, Vania Estivalete, Vivian Flores Costa, Tais De Andrade, Lisiane Fellini Faller

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Social entrepreneurship has emerged and driven a new development perspective, and as the literature mentions, it is based on innovation, and mainly, on the creation of social value, rather than personal wealth and shareholders. In this field of study, one of the focuses of discussion refers to the distinct characteristics of the individuals responsible for socially directed initiatives, named as social entrepreneurs. To contribute to this perspective, the present study aims to identify the values related to work that guide the performance of social entrepreneurs, members of enterprises that have developed themselves within a social incubator at a federal institution of higher education in Brazil. Each person's value system is present in different facets of his life, manifesting himself in his choices and in the way he conducts the relationship with other people in society. Especially the values of work, the focus of this research, play a significant role in organizational studies, since they are considered one of the important guiding principles of the behavior of individuals in the work environment. Regarding the method of the study, a descriptive and qualitative research was carried out. In the data collection, 24 entrepreneurs, members of five different enterprises belonging to the social incubator, were interviewed. The research instrument consisted of three open questions, which could be answered with the support of a "disc of values", an artifact organized to clearly demonstrate the values of the work to the respondents. The analysis of the interviews took into account the categories defined a priori, based on the model proposed by previous authors who validated these constructs within their research contexts, contemplating the following dimensions: Self-determination and stimulation; Safety; Conformity; Universalism and benevolence; Achievement; and Power. It should be noted that, in order to provide a better understanding of the interviewees, in the "disc of values" used in the research, these dimensions were represented by the objectives that define them, being respectively: Challenge; Financial independence; Commitment; Welfare of others; Personal success; And Power. Some preliminary results show that, as guiding principles of the investigation, priority is given to work values related to Self-determination and stimulation, Conformity and Universalism and benevolence. Such findings point to the importance given by these individuals to independent thinking and acting, as well as to novelty and constant challenge. Still, they demonstrate the appreciation of commitment to their enterprise, the people who make it and the quality of their work. They also point to the relevance of the possibility of contributing to the greater social good, that is, of the search for the well-being of close people and of society, as it is implied in models of social entrepreneurship coming from literature. With a lower degree of priority, the values denominated Safety and Realization, as the financial question at work and the search for satisfaction and personal success, through the use of socially recognized skills were mentioned aspects with little emphasis by social entrepreneurs. The Power value was not considered as guiding principle of the work for the respondents.

Keywords: qualitative study, social entrepreneur, social incubator, values of work

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Authors: António J. Gano, Carmen Rangel

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Currently, electric energy storage systems for stationary applications have known an increasing interest, namely with the integration of local renewable energy power sources into energy communities. Li-ion batteries are considered the leading electric storage devices to achieve this integration, and Battery Management Systems (BMS) are decisive for their control and optimum performance. In this work, the advancement of a smart BMS (sBMS) prototype with a modular distributed topology is described. The system, still under development, has a distributed architecture with modular characteristics to operate with different battery pack topologies and charge capacities, integrating adaptive algorithms for functional state real-time monitoring and management of multicellular Li-ion batteries, and is intended for application in the context of a local energy community fed by renewable energy sources. This sBMS system includes different developed hardware units: (1) Cell monitoring units (CMUs) for interfacing with each individual cell or module monitoring within the battery pack; (2) Battery monitoring and switching unit (BMU) for global battery pack monitoring, thermal control and functional operating state switching; (3) Main management and local control unit (MCU) for local sBMS’s management and control, also serving as a communications gateway to external systems and devices. This architecture is fully expandable to battery packs with a large number of cells, or modules, interconnected in series, as the several units have local data acquisition and processing capabilities, communicating over a standard CAN bus and will be able to operate almost autonomously. The CMU units are intended to be used with Li-ion cells but can be used with other cell chemistries, with output voltages within the 2.5 to 5 V range. The different unit’s characteristics and specifications are described, including the different implemented hardware solutions. The developed hardware supports both passive and active methods for charge equalization, considered fundamental functionalities for optimizing the performance and the useful lifetime of a Li-ion battery package. The functional characteristics of the different units of this sBMS system, including different process variables data acquisition using a flexible set of sensors, can support the development of custom algorithms for estimating the parameters defining the functional states of the battery pack (State-of-Charge, State-of-Health, etc.) as well as different charge equalizing strategies and algorithms. This sBMS system is intended to interface with other systems and devices using standard communication protocols, like those used by the Internet of Things. In the future, this sBMS architecture can evolve to a fully decentralized topology, with all the units using Wi-Fi protocols and integrating a mesh network, making unnecessary the MCU unit. The status of the work in progress is reported, leading to conclusions on the system already executed, considering the implemented hardware solution, not only as fully functional advanced and configurable battery management system but also as a platform for developing custom algorithms and optimizing strategies to achieve better performance of electric energy stationary storage devices.

Keywords: Li-ion battery, smart BMS, stationary electric storage, distributed BMS

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Authors: Marta Malo, Carlota Soto, Carmen García-Rosales, Teresa Hernández

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SiC based composites are candidates for possible use as structural and functional materials in the future fusion reactors, the main role is intended for the blanket modules. In the blanket, the neutrons produced in the fusion reaction slow down and their energy is transformed into heat in order to finally generate electrical power. In the blanket design named Dual Coolant Lead Lithium (DCLL), a PbLi alloy for power conversion and tritium breeding circulates inside hollow channels called Flow Channel Inserts (FCIs). These FCI must protect the steel structures against the highly corrosive PbLi liquid and the high temperatures, but also provide electrical insulation in order to minimize magnetohydrodynamic interactions of the flowing liquid metal with the high magnetic field present in a magnetically confined fusion environment. Due to their nominally high temperature and radiation stability as well as corrosion resistance, SiC is the main choice for the flow channel inserts. The significantly lower manufacturing cost presents porous SiC (dense coating is required in order to assure protection against corrosion and as a tritium barrier) as a firm alternative to SiC/SiC composites for this purpose. This application requires the materials to be exposed to high radiation levels and extreme temperatures, conditions for which previous studies have shown noticeable changes in both the microstructure and the electrical properties of different types of silicon carbide. Both initial properties and radiation/temperature induced damage strongly depend on the crystal structure, polytype, impurities/additives that are determined by the fabrication process, so the development of a suitable material requires full control of these variables. For this work, several SiC samples with different percentage of porosity and sintering additives have been manufactured by the so-called sacrificial template method at the Ceit-IK4 Technology Center (San Sebastián, Spain), and characterized at Ciemat (Madrid, Spain). Electrical conductivity was measured as a function of temperature before and after irradiation with 1.8 MeV electrons in the Ciemat HVEC Van de Graaff accelerator up to 140 MGy (~ 2·10 -5 dpa). Radiation-induced conductivity (RIC) was also examined during irradiation at 550 ºC for different dose rates (from 0.5 to 5 kGy/s). Although no significant RIC was found in general for any of the samples, electrical conductivity increase with irradiation dose was observed to occur for some compositions with a linear tendency. However, first results indicate enhanced radiation resistance for coated samples. Preliminary thermogravimetric tests of selected samples, together with posterior XRD analysis allowed interpret radiation-induced modification of the electrical conductivity in terms of changes in the SiC crystalline structure. Further analysis is needed in order to confirm this.

Keywords: DCLL blanket, electrical conductivity, flow channel insert, porous SiC, radiation damage, thermal stability

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Authors: Andrey A. Tyuftin, David Clarke, Malco C. Cruz-Romero, Declan Bolton, Seamus Fanning, Shashi K. Pankaj, Carmen Bueno-Ferrer, Patrick J. Cullen, Joe P. Kerry

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Prolonging of shelf-life is essential in order to address issues such as; supplier demands across continents, economical profit, customer satisfaction, and reduction of food wastage. Smart packaging solutions presented in the form of naturally occurred antimicrobially-active packaging may be a solution to these and other issues. Gelatin film forming solution with adding of natural sourced antimicrobials is a promising tool for the active smart packaging. The objective of this study was to coat conventional plastic hydrophobic packaging material with hydrophilic antimicrobial active beef gelatin coating and conduct shelf life trials on beef sub-primal cuts. Minimal inhibition concentration (MIC) of Caprylic acid sodium salt (SO) and commercially available Auranta FV (AFV) (bitter oranges extract with mixture of nutritive organic acids) were found of 1 and 1.5 % respectively against bacterial strains Bacillus cereus, Pseudomonas fluorescens, Escherichia coli, Staphylococcus aureus and aerobic and anaerobic beef microflora. Therefore SO or AFV were incorporated in beef gelatin film forming solution in concentration of two times of MIC which was coated on a conventional plastic LDPE/PA film on the inner cold plasma treated polyethylene surface. Beef samples were vacuum packed in this material and stored under chilling conditions, sampled at weekly intervals during 42 days shelf life study. No significant differences (p < 0.05) in the cook loss was observed among the different treatments compared to control samples until the day 29. Only for AFV coated beef sample it was 3% higher (37.3%) than the control (34.4 %) on the day 36. It was found antimicrobial films did not protect beef against discoloration. SO containing packages significantly (p < 0.05) reduced Total viable bacterial counts (TVC) compared to the control and AFV samples until the day 35. No significant reduction in TVC was observed between SO and AFV films on the day 42 but a significant difference was observed compared to control samples with a 1.40 log of bacteria reduction on the day 42. AFV films significantly (p < 0.05) reduced TVC compared to control samples from the day 14 until the day 42. Control samples reached the set value of 7 log CFU/g on day 27 of testing, AFV films did not reach this set limit until day 35 and SO films until day 42 of testing. The antimicrobial AFV and SO coated films significantly prolonged the shelf-life of beef steaks by 33 or 55% (on 7 and 14 days respectively) compared to control film samples. It is concluded antimicrobial coated films were successfully developed by coating the inner polyethylene layer of conventional LDPE/PA laminated films after plasma surface treatment. The results indicated that the use of antimicrobial active packaging coated with SO or AFV increased significantly (p < 0.05) the shelf life of the beef sub-primal. Overall, AFV or SO containing gelatin coatings have the potential of being used as effective antimicrobials for active packaging applications for muscle-based food products.

Keywords: active packaging, antimicrobials, edible coatings, food packaging, gelatin films, meat science

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Authors: Katherine Del Carmen Camacho Zorogastua, Victor Hugo Gallo Ramos, Jhon Walter Gomez Lora

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Marine hydrocarbon exploration (oil and natural gas) activities are developed using 2D, 3D and 4D seismic prospecting techniques where sound waves are directed from a seismic vessel emitted every few seconds depending on the variety of air compressors, which cross the layers of rock at the bottom of the sea and are reflected to the surface of the water. Hydrophones receive and record the reflected energy signals for cross-sectional mapping of the lithological profile in order to identify possible areas where hydrocarbon deposits can be formed. However, they produce several significant negative environmental impacts on the marine ecosystem and in the social and economic sectors. Therefore, the objective of the research is to publicize the negative impacts and environmental measures that must be carried out during the development of these activities to prevent and mitigate water quality, the population involved (fishermen) and the marine biota (e.g., Cetaceans, fish) that are the most vulnerable. The research contains technical environmental aspects based on bibliographic sources of environmental studies approved by the Peruvian authority, research articles, undergraduate and postgraduate theses, books, guides, and manuals from Spain, Australia, Canada, Brazil, and Mexico. It describes the negative impacts on the environment and population (fishing sector), environmental prevention, mitigation, recovery and compensation measures that must be properly implemented and the cases of global sea species stranding, for which international experiences from Spain, Madagascar, Mexico, Ecuador, Uruguay, and Peru were referenced. Negative impacts on marine fauna, seawater quality, and the socioeconomic sector (fishermen) were identified. Omission or inadequate biological monitoring in mammals could alter their ability to communicate, feed, and displacement resulting in their stranding and death. In fish, they cause deadly damage to physical-physiological type and in their behavior. Inadequate wastewater treatment and waste management could increase the organic load and oily waste on seawater quality in violation of marine flora and fauna. The possible estrangement of marine resources (fish) affects the economic sector as they carry out their fishing activity for consumption or sale. Finally, it is concluded from the experiences gathered from Spain, Madagascar, Mexico, Ecuador, Uruguay, and Peru that there is a cause and effect relationship between the inadequate development of seismic exploration activities (cause) and marine species strandings (effect) since over the years, stranded or dead marine mammals have been detected on the shores of the sea in areas of seismic acquisition of hydrocarbons. In this regard, it is recommended to establish technical procedures, guidelines, and protocols for the monitoring of marine species in order to contribute to the conservation of hydrobiological resources.

Keywords: 3D seismic prospecting, cetaceans, significant environmental impacts, prevention, mitigation, recovery, environmental compensation

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Authors: Carmen W. H. Chan, Helen Y. L. Chan, Kai Chow Choi, Ka Ming Chow, Cecilia W. M. Kwan, Nancy H. Y. Ng, Jackie Robinson

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Introduction: In Hong Kong, a significant number of deaths occur in acute care wards, which requires nurses in these settings to provide end-of-life care and lead ACP implementation. However, nurses in these settings, in fact, have very low-level involvement in ACP discussions because of limited training in ACP conversations. Objective: This study aims to assess the impact of a multi-media experiential ACP (MEACP) training program, which is guided by the experiential learning model and theory of planned behaviour, on nurses' knowledge and confidence in assisting patients with ACP. Methodology: The study utilizes a cluster randomized controlled trial with a 12-week follow-up. Eligible nurses working in acute care hospital wards are randomly assigned at the ward level, in a 1:1 ratio, to either the control group (no ACP education) or the intervention group (4-week MEACP training program). The training programme includes training through a webpage and mobile application, as well as a face-to-face training workshop with enhanced lectures and role play, which is based on the Theory of Planned Behavior and Kolb's Experiential Learning Model. Questionnaires were distributed to assess nurses' knowledge (a 10-item true/false questionnaire) and level of confidence (five-point Likert scale) in ACP at baseline (T0), four weeks after the baseline assessment (T1), and 12 weeks after T1 (T2). In this interim report, data analysis was mainly descriptive in nature. Result: The interim report focuses on the preliminary results of 165 nurses at T0 (Control: 74, Intervention: 91) over a 5-month period, 69 nurses from the control group who completed the 4-week follow-up and 65 nurses from the intervention group who completed the 4-week MEACP training program at T1. The preliminary attrition rate is 6.8% and 28.6% for the control and intervention groups, respectively, as some nurses did not complete the whole set of online modules. At baseline, the two groups were generally homogeneous in terms of their years of nursing practice, weekly working hours, working title, and level of education, as well as ACP knowledge and confidence levels. The proportion of nurses who answered all ten knowledge questions correctly increased from 13.8% (T0) to 66.2% (T1) for the intervention group and from 13% (T0) to 20.3% (T1) for the control group. The nurses in the intervention group answered an average of 7.57 and 9.43 questions correctly at T0 and T1, respectively. They showed a greater improvement in the knowledge assessment at T1 with respect to T0 when compared with their counterparts in the control group (mean difference of change score, Δ=1.22). They also exhibited a greater gain in level of confidence at T1 compared to their colleagues in the control group (Δ=0.91). T2 data is yet available. Conclusion: The prevalence of nurses engaging in ACP and their level of knowledge about ACP in Hong Kong is low. The MEACP training program can enrich nurses by providing them with more knowledge about ACP and increasing their confidence in conducting ACP.

Keywords: advance directive, advance care planning, confidence, knowledge, multi-media experiential, randomised control trial

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Authors: Beatriz González-rodrigo, Diego Hidalgo-leiva, Omar Flores, Claudia Germoso, Maribel Jiménez-martínez, Laura Navas-sánchez, Belén Orta, Nicola Tarque, Orlando Hernández- Rubio, Miguel Marchamalo, Juan Gregorio Rejas, Belén Benito-oterino

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Cities facing seismic hazard necessitate detailed risk assessments for effective urban planning and vulnerability identification, ensuring the safety and sustainability of urban infrastructure. Comprehensive studies involving seismic hazard, vulnerability, and exposure evaluations are pivotal for estimating potential losses and guiding proactive measures against seismic events. However, broad-scale traditional risk studies limit consideration of specific local threats and identify vulnerable housing within a structural typology. Achieving precise results at neighbourhood levels demands higher resolution seismic hazard exposure, and vulnerability studies. This research aims to bolster sustainability and safety against seismic disasters in three Central American and Caribbean capitals. It integrates geospatial techniques and artificial intelligence into seismic risk studies, proposing cost-effective methods for exposure data collection and damage prediction. The methodology relies on prior seismic threat studies in pilot zones, utilizing existing exposure and vulnerability data in the region. Emphasizing detailed building attributes enables the consideration of behaviour modifiers affecting seismic response. The approach aims to generate detailed risk scenarios, facilitating prioritization of preventive actions pre-, during, and post-seismic events, enhancing decision-making certainty. Detailed risk scenarios necessitate substantial investment in fieldwork, training, research, and methodology development. Regional cooperation becomes crucial given similar seismic threats, urban planning, and construction systems among involved countries. The outcomes hold significance for emergency planning and national and regional construction regulations. The success of this methodology depends on cooperation, investment, and innovative approaches, offering insights and lessons applicable to regions facing moderate seismic threats with vulnerable constructions. Thus, this framework aims to fortify resilience in seismic-prone areas and serves as a reference for global urban planning and disaster management strategies. In conclusion, this research proposes a comprehensive framework for seismic risk assessment in high-risk urban areas, emphasizing detailed studies at finer resolutions for precise vulnerability evaluations. The approach integrates regional cooperation, geospatial technologies, and adaptive fragility curve adjustments to enhance risk assessment accuracy, guiding effective mitigation strategies and emergency management plans.

Keywords: assessment, behaviour modifiers, emergency management, mitigation strategies, resilience, vulnerability

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Authors: Elena Ionescu, Elena Iacob, Marie-Louise Ionescu, Carmen Elena Tebrencu, Oana Teodora Ciuperca

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Echinacea species (Asteraceae) has received a global attention because it is widely used for treatment of cold, flu and upper respiratory tract infections. Echinacea species contain a great variety of chemical components that contribute to their activity. The most important components responsible for the biological activity are those with high molecular-weight such as polysaccharides, polyacetylenes, highly unsaturated alkamides and caffeic acid derivatives. The principal factors that may influence the chemical composition of Echinacea include the species and the part of plant used (aerial parts or roots ). In recent years the market for Echinacea has grown rapidly and also the cases of adultery/replacement especially for Echinacea root. The identification of presence or absence of same biomarkers provide information for safe use of Echinacea species in food supplements industry. The aim of the study was the preliminary evaluation and fingerprinting by UV-VISIBLE spectroscopy of biomarkers in terms of content in phenolic derivatives of some Echinacea species (E. purpurea, E. angustifolia and E. pallida) for identification and authentication of the species. The steps of the study were: (1) samples (extracts) preparation from Echinacea species (non-hydrolyzed and hydrolyzed ethanol extracts); (2) samples preparation of reference substances (polyphenol acids: caftaric acid, caffeic acid, chlorogenic acid, ferulic acid; flavonoids: rutoside, hyperoside, isoquercitrin and their aglycones: quercitri, quercetol, luteolin, kaempferol and apigenin); (3) identification of specific absorption at wavelengths between 700-200 nm; (4) identify the phenolic compounds from Echinacea species based on spectral characteristics and the specific absorption; each class of compounds corresponds to a maximum absorption in the UV spectrum. The phytochemical compounds were identified at specific wavelengths between 700-200 nm. The absorption intensities were measured. The obtained results proved that ethanolic extract showed absorption peaks attributed to: phenolic compounds (free phenolic acids and phenolic acids derivatives) registrated between 220-280 nm, unsymmetrical chemical structure compounds (caffeic acid, chlorogenic acid, ferulic acid) with maximum absorption peak and absorption "shoulder" that may be due to substitution of hydroxyl or methoxy group, flavonoid compounds (in free form or glycosides) between 330-360 nm, due to the double bond in position 2,3 and carbonyl group in position 4 flavonols. UV spectra showed two major peaks of absorption (quercetin glycoside, rutin, etc.). The results obtained by UV-VIS spectroscopy has revealed the presence of phenolic derivatives such as cicoric acid (240 nm), caftaric acid (329 nm), caffeic acid (240 nm), rutoside (205 nm), quercetin (255 nm), luteolin (235 nm) in all three species of Echinacea. The echinacoside is absent. This profile mentioned above and the absence of phenolic compound echinacoside leads to the conclusion that species harvested as Echinacea angustifolia and Echinacea pallida are Echinacea purpurea also; It can be said that preliminary fingerprinting of Echinacea species through correspondence with the phenolic derivatives profile can be achieved by UV-VIS spectroscopic investigation, which is an adequate technique for preliminary identification and authentication of Echinacea in medicinal herbs.

Keywords: Echinacea species, Fingerprinting, Phenolic compounds, UV-VIS spectroscopy

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Authors: Carmen Axisa, Louise Nash, Patrick Kelly, Simon Willcock

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Introduction: Doctors experience high levels of burnout, stress and psychiatric morbidity. This can affect the health of the doctor and impact patient care. Study Aims: To evaluate the effectiveness of a workshop intervention to promote wellbeing for Australian Physician Trainees. Methods: A workshop was developed in consultation with specialist clinicians to promote health and wellbeing for physician trainees. The workshop objectives were to improve participant understanding about factors affecting their health and wellbeing, to outline strategies on how to improve health and wellbeing and to encourage participants to apply these strategies in their own lives. There was a focus on building resilience and developing long term healthy behaviours as part of the physician trainee daily lifestyle. Trainees had the opportunity to learn practical strategies for stress management, gain insight into their behaviour and take steps to improve their health and wellbeing. The workshop also identified resources and support systems available to trainees. The workshop duration was four and a half hours including a thirty- minute meal break where a catered meal was provided for the trainees. Workshop evaluations were conducted at the end of the workshop. Sixty-seven physician trainees from Adult Medicine and Paediatric training programs in Sydney Australia were randomised into intervention and control groups. The intervention group attended a workshop facilitated by specialist clinicians and the control group did not. Baseline and post intervention measurements were taken for both groups to evaluate the impact and effectiveness of the workshop. Forty-six participants completed all three measurements (69%). Demographic, personal and self-reported data regarding work/life patterns was collected. Outcome measures include Depression Anxiety Stress Scale (DASS), Professional Quality of Life Scale (ProQOL) and Alcohol Use Disorders Identification Test (AUDIT). Results: The workshop was well received by the physician trainees and workshop evaluations showed that the majority of trainees strongly agree or agree that the training was relevant to their needs (96%) and met their expectations (92%). All trainees strongly agree or agree that they would recommend the workshop to their medical colleagues. In comparison to the control group we observed a reduction in alcohol use, depression and burnout but an increase in stress, anxiety and secondary traumatic stress in the intervention group, at the primary endpoint measured at 6 months. However, none of these differences reached statistical significance (p > 0.05). Discussion: Although the study did not reach statistical significance, the workshop may be beneficial to physician trainees. Trainees had the opportunity to share ideas, gain insight into their own behaviour, learn practical strategies for stress management and discuss approach to work, life and self-care. The workshop discussions enabled trainees to share their experiences in a supported environment where they learned that other trainees experienced stress and burnout and they were not alone in needing to acquire successful coping mechanisms and stress management strategies. Conclusion: These findings suggest that physician trainees are a vulnerable group who may benefit from initiatives that promote wellbeing and from a more supportive work environment.

Keywords: doctors' health, physician burnout, physician resilience, wellbeing workshop

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Authors: Richard McGregor, Carmen Reaiche, Stephen Boyle

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Cyber threats are a relatively recent phenomenon and offer cyber insurers a dynamic and intelligent peril. As individuals en mass become increasingly digitally dependent, Personal Cyber Insurance (PCI) offers an attractive option to mitigate cyber risk at a personal level. This abstract proposes a literature review that conceptualises a framework for siting Personal Cyber Insurance (PCI) within the context of cyberspace. The lack of empirical research within this domain demonstrates an immediate need to define the scope of PCI to allow cyber insurers to understand personal cyber risk threats and vectors, customer awareness, capabilities, and their associated needs. Additionally, this will allow cyber insurers to conceptualise appropriate frameworks allowing effective management and distribution of PCI products and services within a landscape often in-congruent with risk attributes commonly associated with traditional personal line insurance products. Cyberspace has provided significant improvement to the quality of social connectivity and productivity during past decades and allowed enormous capability uplift of information sharing and communication between people and communities. Conversely, personal digital dependency furnish ample opportunities for adverse cyber events such as data breaches and cyber-attacksthus introducing a continuous and insidious threat of omnipresent cyber risk–particularly since the advent of the COVID-19 pandemic and wide-spread adoption of ‘work-from-home’ practices. Recognition of escalating inter-dependencies, vulnerabilities and inadequate personal cyber behaviours have prompted efforts by businesses and individuals alike to investigate strategies and tactics to mitigate cyber risk – of which cyber insurance is a viable, cost-effective option. It is argued that, ceteris parabus, the nature of cyberspace intrinsically provides characteristic peculiarities that pose significant and bespoke challenges to cyber insurers, often in-congruent with risk attributes commonly associated with traditional personal line insurance products. These challenges include (inter alia) a paucity of historical claim/loss data for underwriting and pricing purposes, interdependencies of cyber architecture promoting high correlation of cyber risk, difficulties in evaluating cyber risk, intangibility of risk assets (such as data, reputation), lack of standardisation across the industry, high and undetermined tail risks, and moral hazard among others. This study proposes a thematic overview of the literature deemed necessary to conceptualise the challenges to issuing personal cyber coverage. There is an evident absence of empirical research appertaining to PCI and the design of operational business models for this business domain, especially qualitative initiatives that (1) attempt to define the scope of the peril, (2) secure an understanding of the needs of both cyber insurer and customer, and (3) to identify elements pivotal to effective management and profitable distribution of PCI - leading to an argument proposed by the author that postulates that the traditional general insurance customer journey and business model are ill-suited for the lineaments of cyberspace. The findings of the review confirm significant gaps in contemporary research within the domain of personal cyber insurance.

Keywords: cyberspace, personal cyber risk, personal cyber insurance, customer journey, business model

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Authors: Carmen García Sánchez

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The UN's 17 Sustainable Development Goals – specifically the nº 3 and nº 11- urgently call for new architectural design solutions at different design scales to increase human contact with nature in the health and wellbeing promotion of primarily urban communities. The discipline of Interior Design offers an important alternative to large-scale nature-inclusive actions which are not always possible due to space limitations. These circumstances provide an immense opportunity to integrate biophilic design, a complex emerging and under-developed approach that pursues sustainable design strategies for increasing the human-nature connection through the experience of the built environment. Biophilic design explores the diverse ways humans are inherently inclined to affiliate with nature, attach meaning to and derive benefit from the natural world. It represents a biological understanding of architecture which categorization is still in progress. The internationally renowned Danish domestic architecture built in the 1950´s and early 1960´s - a golden age of Danish modern architecture - left a leading legacy that has greatly influenced the domestic sphere and has further led the world in terms of good design and welfare. This study examines how four existing post-war domestic buildings establish a dialogue with nature and her variations over time. The case-studies unveil both memorable and unique biophilic resources through sophisticated and original design expressions, where transformative processes connect the users to the natural setting and reflect fundamental ways in which they attach meaning to the place. In addition, fascinating analogies in terms of this nature interaction with particular traditional Japanese architecture inform the research. They embody prevailing lessons for our time today. The research methodology is based on a thorough literature review combined with a phenomenological analysis into how these case-studies contribute to the connection between humans and nature, after conducting fieldwork throughout varying seasons to document understanding in nature transformations multi-sensory perception (via sight, touch, sound, smell, time and movement) as a core research strategy. The cases´ most outstanding features have been studied attending the following key parameters: 1. Space: 1.1. Relationships (itineraries); 1.2. Measures/scale; 2. Context: Context: Landscape reading in different weather/seasonal conditions; 3. Tectonic: 3.1. Constructive joints, elements assembly; 3.2. Structural order; 4. Materiality: 4.1. Finishes, 4.2. Colors; 4.3. Tactile qualities; 5. Daylight interplay. Departing from an artistic-scientific exploration this groundbreaking study provides sustainable practical design strategies, perspectives, and inspiration to boost humans´ contact with nature through the experience of the interior built environment. Some strategies are associated with access to outdoor space or require ample space, while others can thrive in a dense urban context without direct access to the natural environment. The objective is not only to produce knowledge, but to phase in biophilic design in the built environment, expanding its theory and practice into a new dimension. Its long-term vision is to efficiently enhance the health and well-being of urban communities through daily interaction with Nature.

Keywords: sustainability, biophilic design, architectural design, interior design, nature, Danish architecture, Japanese architecture

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Authors: Elena Iacob, Marie-Louise Ionescu, Elena Ionescu, Carmen Elena Tebrencu, Oana Teodora Ciuperca

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Infrared spectroscopy (FT-IR) is an advanced technique frequently used to authenticate both raw materials and final products using their specific fingerprints and to determine plant extracts biomarkers based on their functional groups. In recent years the market for Hypericum has grown rapidly and also has grown the cases of adultery/replacement, especially for Hypericum perforatum L.specie. Presence/absence of same biomarkers provides preliminary identification of Hypericum species in safe use in the manufacture of food supplements. The main objective of the work was to characterize the main biomarkers of Hypericum perforatum L. (St. John's wort) and identify this species in herbal food supplements after specific FT-IR fingerprint. An experimental program has been designed in order to test: (1) raw material (St. John's wort); (2)intermediate raw materials (St. John's wort dry extract ); (3) the finished products: tablets based on powders, on extracts, on powder and extract, hydroalcoholic solution from herbal mixture based on St. John's wort. The analyze using FTIR infrared spectroscopy were obtained raw materials, intermediates and finished products spectra, respectively absorption bands corresponding and similar with aliphatic and aromatic structures; examination was done individually and through comparison between Hypericum perforatum L. plant species and finished product The tests were done in correlation with phytochemical markers for authenticating the specie Hypericum perforatum L.: hyperoside, rutin, quercetin, isoquercetin, luteolin, apigenin, hypericin, hyperforin, chlorogenic acid. Samples were analyzed using a Shimatzu FTIR spectrometer and the infrared spectrum of each sample was recorded in the MIR region, from 4000 to 1000 cm-1 and then the fingerprint region was selected for data analysis. The following functional groups were identified -stretching vibrations suggests existing groups in the compounds of interest (flavones–rutin, hyperoside, polyphenolcarboxilic acids - chlorogenic acid, naphtodianthrones- hypericin): oxidril groups (OH) free alcohol type: rutin, hyperoside, chlorogenic acid; C = O bond from structures with free carbonyl groups of aldehyde, ketone, carboxylic, ester: hypericin; C = O structure with the free carbonyl of the aldehyde groups, ketone, carboxylic acid, esteric/C = O free bonds present in chlorogenic acid; C = C bonds of the aromatic ring (condensed aromatic hydrocarbons, heterocyclic compounds) present in all compounds of interest; OH phenolic groups: present in all compounds of interest, C-O-C groups from glycoside structures: rutin, hyperoside, chlorogenic acid. The experimental results show that: (I)The six fingerprint region analysis indicated the presence of specific functional groups: (1) 1000 - 1130 cm-1 (C-O–C of glycoside structures); (2) 1200-1380 cm-1 (carbonyl C-O or O-H phenolic); (3) 1400-1450 cm-1 (C=C aromatic); (4) 1600- 1730 cm-1 (C=O carbonyl); (5) 2850 - 2930 cm-1 (–CH3, -CH2-, =CH-); (6) 338-3920 cm-1 (OH free alcohol type); (II)Comparative FT-IR spectral analysis indicate the authenticity of the finished products ( tablets) in terms of Hypericum perforatum L. content; (III)The infrared spectroscopy is an adequate technique for identification and authentication of the medicinal herbs , intermediate raw material and in the food supplements less in the form of solutions where the results are not conclusive.

Keywords: Authentication, FT-IR fingerprint, Herbal supplements, Hypericum perforatum L.

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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

Abstract:

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

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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

Abstract:

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

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