Search results for: Tania Vazquez

Authors: Ebrahim Shayesteh, Nikolaos Styliaras, Alin George Raducu, Ozan Sahin, Daniel Pombo VáZquez, Jonas Funkquist, Sotirios Thanopoulos

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One of the important requirements in many code generation projects is defining some of the model parameters tunable. This helps to update the model parameters without performing the code generation again. This paper studies the concept of embedded code generation by MATLAB/Simulink coder targeting the TwinCAT Simulink system. The generated runtime modules are then tested and deployed to the TwinCAT 3 engineering environment. However, defining the parameters tunable in MATLAB/Simulink code generation targeting TwinCAT is not very straightforward. This paper focuses on this subject and reviews some of the techniques tested here to make the parameters tunable in generated runtime modules. Three techniques are proposed for this purpose, including normal tunable parameters, callback functions, and mask subsystems. Moreover, some test Simulink models are developed and used to evaluate the results of proposed approaches. A brief summary of the study results is presented in the following. First of all, the parameters defined tunable and used in defining the values of other Simulink elements (e.g., gain value of a gain block) could be changed after the code generation and this value updating will affect the values of all elements defined based on the values of the tunable parameter. For instance, if parameter K=1 is defined as a tunable parameter in the code generation process and this parameter is used to gain a gain block in Simulink, the gain value for the gain block is equal to 1 in the gain block TwinCAT environment after the code generation. But, the value of K can be changed to a new value (e.g., K=2) in TwinCAT (without doing any new code generation in MATLAB). Then, the gain value of the gain block will change to 2. Secondly, adding a callback function in the form of “pre-load function,” “post-load function,” “start function,” and will not help to make the parameters tunable without performing a new code generation. This means that any MATLAB files should be run before performing the code generation. The parameters defined/calculated in this file will be used as fixed values in the generated code. Thus, adding these files as callback functions to the Simulink model will not make these parameters flexible since the MATLAB files will not be attached to the generated code. Therefore, to change the parameters defined/calculated in these files, the code generation should be done again. However, adding these files as callback functions forces MATLAB to run them before the code generation, and there is no need to define the parameters mentioned in these files separately. Finally, using a tunable parameter in defining/calculating the values of other parameters through the mask is an efficient method to change the value of the latter parameters after the code generation. For instance, if tunable parameter K is used in calculating the value of two other parameters K1 and K2 and, after the code generation, the value of K is updated in TwinCAT environment, the value of parameters K1 and K2 will also be updated (without any new code generation).

Keywords: code generation, MATLAB, tunable parameters, TwinCAT

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

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

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

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Authors: Alejandra G. Barbón, José Vila, Dania Vazquez

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This study seeks to contribute to the advancement of library and archival sciences in the areas of records management, knowledge organization, and information architecture, particularly focusing on the enhancement of bibliographical description through the incorporation of visual interactive designs aimed to enrich the library users’ experience. In an era of heightened awareness about the legacy of hiddenness across special and rare collections in libraries and archives, along with the need for inclusivity in academia, the University of Miami Libraries has embarked on an innovative project that intersects the realms of print culture, decolonization, and digital technology. This proposal presents an exciting initiative to revitalize the study of Artists’ Books collections by employing digital visual representations to decolonize bibliographic records of some of the most unique materials and foster a more holistic understanding of cultural heritage. Artists' Books, a dynamic and interdisciplinary art form, challenge conventional bibliographic classification systems, making them ripe for the exploration of alternative approaches. This project involves the creation of a digital platform that combines multimedia elements for digital representations, interactive information retrieval systems, innovative information architecture, trending bibliographic cataloging and metadata initiatives, and collaborative curation to transform how we engage with and understand these collections. By embracing the potential of technology, we aim to transcend traditional constraints and address the historical biases that have influenced bibliographic practices. In essence, this study showcases a groundbreaking endeavor at the University of Miami Libraries that seeks to not only enhance bibliographic practices but also confront the legacy of hiddenness across special and rare collections in libraries and archives while strengthening conventional bibliographic description. By embracing digital visualizations, we aim to provide new pathways for understanding Artists' Books collections in a manner that is more inclusive, dynamic, and forward-looking. This project exemplifies the University’s dedication to fostering critical engagement, embracing technological innovation, and promoting diverse and equitable classifications and representations of cultural heritage.

Keywords: decolonizing bibliographic cataloging frameworks, digital visualizations information architecture platforms, collaborative curation and inclusivity for records management, engagement and accessibility increasing interaction design and user experience

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Authors: Sandra Figueiredo, Alexandra Pereira, Ana Oliveira, Idia Brito, Ivaniltan Jones, Joana Moreira, Madalena Silva, Maria Paraíba, Milene Silva, Tânia Pinho

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In the present society, we are facing behaviours mainly in young individuals that might be considered trends of culture or psychopathology. Both contexts are challenges for Education, Psychology and Health. This paper examines nine case studies specifically in Educational Psychology with the main goal to identify and define phenomena contexts in school culture, the psychopathology involved and to present a psychological intervention for each case. The research was conducted by university students in the period of March 2017-June 2017, in Portugal, and the childhood was focused. The case studies explored the cyberbullying; the bullying - victims and bullies’ perspectives; the obsessive compulsive disorder; perception and inclusion of children from homoparental families; inclusion of foreign students in the higher education system; blindness and the inclusion in physical curricular activities; influence of doc-reality and media in attitudes and self-esteem; and the morningness and eveningness types learning in the same school timetables. The university students were supervised during their research analysis and two methods were available for the intervention research study: the meta-analysis and the empirical study. In the second phase, the pedagogical intervention was designed for the different educational contexts in analysis, especially concerning the school environments. The evidence of literature and the empirical studies showed new trends of school’ behaviours and educational disturbances that require further research and effective (and adequate to age, gender, nationality and culture) pedagogical instruments. Respecting the instruments, on the one hand, to identify behaviors, habits or pathologies and highlight the role and training of teachers, psychologists and health professionals, on the other hand, to promote the early intervention and to enhance healthy child development and orientation of the families. To respond to both milestones, this paper present nine pedagogical techniques and measures that will be discussed on their impact concerning advances for the psychological and educational intervention, centered in the individual and in the new generations of family’ cultures.

Keywords: behaviour, culture trends, educational intervention, psychopathology, obsessive compulsive disorder, cyberbullying, bullying, homoparental families, sleep influence, blindness and sports at school, inclusion of foreign students, media influence in behaviour

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Authors: Tania Lisboa, Angela Voyajolu, Adam Ockelford

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The positive impact of music on the development of children with autism is widely acknowledged: music offers a unique channel for communication, wellbeing and self-regulation, as well as access to culture and a means of creative engagement. Yet, no coherent program exists for parents, carers and teachers to follow with their children in the early years, when the need for interventions is often most acute. Hence, research and the development of resources is urgently required. Autism is a project with children on the autism spectrum. The project aims at promoting the participants’ engagement with music through involvement in specially-designed musical activities with parents and carers. The main goal of the research is to verify the effectiveness of newly designed resources and strategies, which are based on the Sounds of Intent in the Early Years (SoI-EY) framework of musical development. This is a pilot study, comprising case studies of five children with autism in the early years. The data comprises semi-structured interviews, observations of videos, and feedback from parents on resources. Interpretative Phenomenological Analysis was chosen to analyze the interviews. The video data was coded in relation to the SoI-EY framework. The feedback from parents was used to evaluate the resources (i.e. musical activity cards). The participants’ wider development was also assessed through selected elements of the Early Years Foundation Stage (EYFS), a national assessment framework used in England: specifically, communication, language and social-emotional development. Five families of children on the autism spectrum (aged between 4-8 years) participated in the pilot. The research team visited each family 4 times over a 3-month period, during which the children were observed, and musical activities were suggested based on the child’s assessed level of musical development. Parents then trialed the activities, providing feedback and gathering further video observations of their child’s musical engagement between visits. The results of one case study will be featured in this paper, in which the evidence suggests that specifically tailored musical activity may promote communication and social engagement for a child with language difficulties on the autism spectrum. The resources were appropriate for the children’s involvement in musical activities. Findings suggest that non-specialist musical engagement with family and carers can be a powerful means to foster communication. The case study featured in this paper illustrates this with a child of limited verbal ability. There is a need for further research and development of resources that can be made available to all those working with children on the autism spectrum.

Keywords: autism, development, music education, resources

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Authors: Tania Chalton

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In June 2013, an independent review of the Liverpool Care Pathway (LCP) identified a number of problems with the implementation of the LCP in the UK and recommended that it be replaced by individual care plans for each patient. As a result of the UK findings, in November 2013 the Ministry of Health (MOH) commissioned the Palliative Care Council to initiate a programme of work to investigate an appropriate approach for the care of people in their last days of life in New Zealand (NZ). The Last Days of Life Working Group commenced a process to develop national consensus on the care of people in their last days of life in April 2014. In order to develop its advice for the future provision of care to people in their last days of life, the Working Group (WG) established a comprehensive work programme and as a result has developed a series of working papers. Specific areas of focus included: An analysis of the UK Independent Review findings and an assessment of these findings to the NZ context. A stocktake of services providing care to people in their last days of life, including aged residential care (ARC); hospices; hospitals; and primary care. International and NZ literature reviews of evidence and best practice. Survey of family to understand the consumer perspective on the care of people in their last days of life. Key aspects of care that required further considerations for NZ were: Terminology: clarify terminology used in the last days of life and in relation to death and dying. Evidenced based: including specific review of evidence regarding, spiritual, culturally appropriate care as well as dementia care. Diagnosis of dying: need for both guidance around the diagnosis of dying and communication with family. Workforce issues: access to an appropriate workforce after hours. Nutrition and hydration: guidance around appropriate approaches to nutrition and hydration. Symptom and pain management: guidance around symptom management. Documentation: documentation of the person’s care which is robust enough for data collection and auditing requirements, not ‘tick box’ approach to care. Education and training: improved consistency and access to appropriate education and training. Leadership: A dedicated team or person to support and coordinate the introduction and implementation of any last days of life model of care. Quality indicators and data collection: model of care to enable auditing and regular reviews to ensure on-going quality improvement. Cultural and spiritual: address and incorporate any cultural and spiritual aspects. A final document was developed incorporating all the evidence which provides guidance to the health sector on best practice for people at end of life: “Principles and guidance for the last days of life: Te Ara Whakapiri”.

Keywords: end of life, guidelines, New Zealand, palliative care

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Authors: B. Gonzalez-Yebra, H. Barajas, P. Palomares, M. Hernandez, O. Torres, M. Ayala, A. L. González, G. Vazquez-Ortiz, M. L. Guzman

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Leukemia arises by molecular alterations of the normal hematopoietic stem cell (HSC) transforming it into a leukemic stem cell (LSC) with high cell proliferation, self-renewal, and cell differentiation. Chronic myeloid leukemia (CML) originates from an LSC-leading to elevated proliferation of myeloid cells and acute lymphoblastic leukemia (ALL) originates from an LSC development leading to elevated proliferation of lymphoid cells. In both cases, LSC can be identified by multicolor flow cytometry using several antibodies. However, to date, LSC levels in peripheral blood (PB) are not established well enough in ALL and CML patients. On the other hand, the detection of the minimal residue disease (MRD) in leukemia is mainly based on the identification of the mRNA bcr-abl gene in CML patients and some other genes in ALL patients. There is no a properly biomarker to detect MDR in both types of leukemia. The objective of this study was to determine mRNA bcr-abl and the percentage of LSC in peripheral blood of patients with CML and ALL and identify a possible association between the amount of LSC in PB and clinical data. We included in this study 19 patients with Leukemia. A PB sample was collected per patient and leukocytes were obtained by Ficoll gradient. The immunophenotype for LSC CD34+ was done by flow cytometry analysis with CD33, CD2, CD14, CD16, CD64, HLA-DR, CD13, CD15, CD19, CD10, CD20, CD34, CD38, CD71, CD90, CD117, CD123 monoclonal antibodies. In addition, to identify the presence of the mRNA bcr-abl by RT-PCR, the RNA was isolated using TRIZOL reagent. Molecular (presence of mRNA bcr-abl and LSC CD34+) and clinical results were analyzed with descriptive statistics and a multiple regression analysis was performed to determine statistically significant association. In total, 19 patients (8 patients with ALL and 11 patients with CML) were analyzed, 9 patients with de novo leukemia (ALL = 6 and CML = 3) and 10 under treatment (ALL = 5 and CML = 5). The overall frequency of mRNA bcr-abl was 31% (6/19), and it was negative in ALL patients and positive in 80% in CML patients. On the other hand, LSC was determined in 16/19 leukemia patients (%LSC= 0.02-17.3). The Novo patients had higher percentage of LSC (0.26 to 17.3%) than patients under treatment (0 to 5.93%). The amount of LSC was significantly associated with the amount of LSC were: absence of treatment, the absence of splenomegaly, and a lower number of leukocytes, negative association for the clinical variables age, sex, blasts, and mRNA bcr-abl. In conclusion, patients with de novo leukemia had a higher percentage of circulating LSC than patients under treatment, and it was associated with clinical parameters as lack of treatment, absence of splenomegaly and a lower number of leukocytes. The mRNA bcr-abl detection was only possible in the series of patients with CML, and molecular detection of LSC could be identified in the peripheral blood of all leukemia patients, we believe the identification of circulating LSC may be used as biomarker for the detection of the MRD in leukemia patients.

Keywords: stem cells, leukemia, biomarkers, flow cytometry

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Authors: Pauline Blanchet

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In a world where images are a prominent part of our daily lives and a way of absorbing information, the analysis of the representation of migration narratives is vital. This thesis raises questions concerning the power of illustrations, drawings and visual culture in order to represent the migration narratives in the age of Instagram. The rise of graphic novels and comics has come about in the last fifteen years, specifically regarding contemporary authors engaging with complex social issues such as migration and refugeehood. Due to this, refugee subjects are often in these narratives, whether they are autobiographical stories or whether the subject is included in the creative process. Growth in discourse around migration has been present in other art forms; in 2018, there has been dedicated exhibitions around migration such as Tania Bruguera at the TATE (2018-2019), ‘Journeys Drawn’ at the House of Illustration (2018-2019) and dedicated film festivals (2018; the Migration Film Festival), which have shown the recent considerations of using the arts as a medium of expression regarding themes of refugeehood and migration. Graphic visuals are fast becoming a key instrument when representing migration, and the central thesis of this paper is to show the strength and limitations of this form as well the methodology used by the actors in the production process. Recent works which have been released in the last ten years have not being analysed in the same context as previous graphic novels such as Palestine and Persepolis. While a lot of research has been done on the mass media portrayals of refugees in photography and journalism, there is a lack of literature on the representation with illustrations. There is little research about the accessibility of graphic novels such as where they can be found and what the intentions are when writing the novels. It is interesting to see why these authors, NGOs, and curators have decided to highlight these migrant narratives in a time when the mainstream media has done extensive coverage on the ‘refugee crisis’. Using primary data by doing one on one interviews with artists, curators, and NGOs, this paper investigates the efficiency of graphic novels for depicting refugee stories as a viable alternative to other mass medium forms. The paper has been divided into two distinct sections. The first part is concerned with the form of the comic itself and how it either limits or strengthens the representation of migrant narratives. This will involve analysing the layered and complex forms that comics allow such as multimedia pieces, use of photography and forms of symbolism. It will also show how the illustration allows for anonymity of refugees, the empathetic aspect of the form and how the history of the graphic novel form has allowed space for positive representations of women in the last decade. The second section will analyse the creative and methodological process which takes place by the actors and their involvement with the production of the works.

Keywords: graphic novel, refugee, communication, media, migration

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Authors: Estefanya Vazquez-Casaubon, Veroline Cauberghe, Dieneke Van de Sompel, Hayley Pearce

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Working together to reduce the anthropogenic impact should be a collective action, including effort within the household. In the matter, children are considered to have an important role in influencing the household to reduce the environmental impact through reversed socialization where children motivate and increase the concern of the parents towards environmental protection. Previous studies reveal that communication between parents and kids is key for effective reversed socialization. However, multiple barriers have been identified in the literature, such as the acceptance of the influence from the kids, the properties of the communication, among other factors. Based on the previous evidence, the present study aims to assess barriers and facilitators of communication at the household level that have an impact on reversed socialization. More precisely, the study examines how parents’ climate change coping strategy (problem-focused, meaning-focused, disregarding) influences the valence and the type of the communication related to climate change, and eventually the extent to which they report their beliefs and behaviours to be influenced by the pro-environmental perspectives of their children; i.e. reversed socialization. Via an online survey, 723 Belgian parents self-reported on communication about environmental protection and risk within their household (such as the frequency of exchange about topics related to climate change sourced from school, the household rules, imparting knowledge to the children, and outer factors like media or peer pressure, the emotional valence of the communication), their perceived socialization, and personal factors (coping mechanisms towards climate change). The results, using structural equation modelling, revealed that parents applying a problem-solving coping strategy related to climate change, appear to communicate more often in a positive than in a negative manner. Parents with a disregarding coping style towards climate change appear to communicate less often in a positive way within the household. Parents that cope via meaning-making of climate change showed to communicate less often in either a positive or negative way. Moreover, the perceived valence of the communication (positive or negative) influenced the frequency and type of household communication. Positive emotions increased the frequency of the communication overall. However, the direct effect of neither of the coping mechanisms on the reversed socialization was significant. High frequency of communication about the media, environmental views of the household members among other external topics had a positive impact on the perceived socialization, followed by discussions school-related; while parental instructing had a negative impact on the perceived socialization. Moreover, the frequency of communication was strongly affected by the perceived valence of the communication (positive or negative). The results go in line with previous evidence that a higher frequency of communication facilitates reversed socialization. Hence the results outstand how the coping mechanisms of the parents can be either a facilitator when they cope via problem-solving, while parents that disregard might avert frequent communication about climate change at the household.

Keywords: communication, parents’ coping mechanisms, environmental protection, household, perceived socialization

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Authors: Geraldine G. Granados Vazquez

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Every human being is exposed to the risk of dying; wherein some of them are more susceptible than others depending on the cause. Therefore, the cause could be the hazard to die that a group or individual has, making this irreversible damage the condition of vulnerability. Risk is a dynamic concept; which means that it depends on the environmental, social, economic and political conditions. Thus vulnerability may only be evaluated in terms of relative parameters. This research is focusing specifically on building a model that evaluate the risk or propensity of death in past urban societies in connection with the everyday life of individuals, considering that death can be a consequence of two coexisting issues: hazard and the deterioration of the resistance to destruction. One of the most important discussions in bioarchaeology refers to health and life conditions in ancient groups; the researchers are looking for more flexible models that evaluate these topics. In that way, this research proposes a theoretical-methodological model that assess the vulnerability of death in past urban groups. This model pretends to be useful to evaluate the risk of death, considering their sociohistorical context, and their intrinsic biological features. This theoretical and methodological model, propose four areas to assess vulnerability. The first three areas use statistical methods or quantitative analysis. While the last and fourth area, which corresponds to the embodiment, is based on qualitative analysis. The four areas and their techniques proposed are a) Demographic dynamics. From the distribution of age at the time of death, the analysis of mortality will be performed using life tables. From here, four aspects may be inferred: population structure, fertility, mortality-survival, and productivity-migration, b) Frailty. Selective mortality and heterogeneity in frailty can be assessed through the relationship between characteristics and the age at death. There are two indicators used in contemporary populations to evaluate stress: height and linear enamel hypoplasias. Height estimates may account for the individual’s nutrition and health history in specific groups; while enamel hypoplasias are an account of the individual’s first years of life, c) Inequality. Space reflects various sectors of society, also in ancient cities. In general terms, the spatial analysis uses measures of association to show the relationship between frail variables and space, d) Embodiment. The story of everyone leaves some evidence on the body, even in the bones. That led us to think about the dynamic individual's relations in terms of time and space; consequently, the micro analysis of persons will assess vulnerability from the everyday life, where the symbolic meaning also plays a major role. In sum, using some Mesoamerica examples, as study cases, this research demonstrates that not only the intrinsic characteristics related to the age and sex of individuals are conducive to vulnerability, but also the social and historical context that determines their state of frailty before death. An attenuating factor for past groups is that some basic aspects –such as the role they played in everyday life– escape our comprehension, and are still under discussion.

Keywords: bioarchaeology, frailty, Mesoamerica, vulnerability

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Authors: Karen L. Rincon-Granados, América R. Vázquez-Olmos, Adriana-Patricia Rodríguez-Hernández, Gina Prado-Prone, Margarita Rivera, Roberto Y. Sato-Berrú

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In this work, a hybrid film based on poly-3-hydroxybutyrate (P3HB) and nickel ferrite (NiFe₂O₄) nanoparticles (NPs) was obtained by a simple and reproducible methodology in order to study its antibacterial and cytotoxic properties. The motivation for this research is the current antimicrobial resistance (RAM). This is a threat to human health and development worldwide. RAM is caused by the emergence of bacterial strains resistant to traditional antibiotics that were used as treatment. Due to this, the need to investigate new alternatives for preventing and treating bacterial infections emerges. In this sense, metal oxide NPs have aroused great interest due to their unique physicochemical properties. However, their use is limited by the nanostructured nature, commonly obtained by chemical and physical synthesis methods, as powders or colloidal dispersions. Therefore, the incorporation of nanostructured materials in polymer matrices to obtain hybrid materials that allow disinfecting and preventing the spread of bacteria on various surfaces. Accordingly, this work presents the synthesis and study of the antibacterial properties of the P3HB@NiFe₂O₄ hybrid film as a potential material to inhibit bacterial growth. The NiFe₂O₄ NPs were previously synthesized by a mechanochemical method. The P3HB and P3HB@NiFe₂O₄ films were obtained by the solvent casting method. The films were characterized by X-ray diffraction (XRD), Raman scattering, and scanning electron microscopy (SEM). The XRD pattern showed that the NiFe₂O₄ NPs were incorporated into the P3HB polymer matrix and retained their nanometric sizes. By energy dispersive X-ray spectroscopy (EDS), it was observed that the NPs are homogeneously distributed in the film. The bactericidal effect of the films obtained was evaluated in vitro using the broth surface method against two opportunistic and nosocomial pathogens, Staphylococcus aureus and Pseudomonas aeruginosa. The bacterial growth results showed that the P3HB@NiFe₂O₄ hybrid film was inhibited by 97% and 96% for S. aureus and P. aeruginosa, respectively. Surprisingly, the P3HB film inhibited both bacterial strains by around 90%. The cytotoxicity of the NiFe₂O₄ NPs, P3HB@NiFe₂O₄ hybrid film, and the P3HB film was evaluated using human skin cells, keratinocytes, and fibroblasts, finding that the NPs are biocompatible. The P3HB film and hybrids are cytotoxic, which demonstrated that although P3HB is known and reported as a biocompatible polymer, under our work conditions, P3HB was cytotoxic. Its bactericidal effect could be related to this activity. Its films are bactericidal and cytotoxic to keratinocytes and fibroblasts, the first barrier of human skin. Despite this, the hybrid film of P3HB@NiFe₂O₄ presents synergy with the bactericidal effect between P3HB and NPs, increasing bacterial inhibition. In addition, NPs decrease the cytotoxicity of P3HB to keratinocytes. The methodology used in this work was successful in producing hybrid films with antibacterial activity. However, future challenges are generated to find relationships between NPs and P3HB that allow taking advantage of their bactericidal properties and do not compromise biocompatibility.

Keywords: poly-3-hydroxybutyrate, nanoparticles, hybrid film, antibacterial

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Authors: Britt Erni, Aldara Vazquez Fernandez, Roland Maurer

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Visuospatial perspective taking (VSPT) is a process that allows to integrate spatial information from different points of view, and to transform the mental images we have of the environment to properly orient our movements and anticipate the location of landmarks during navigation. VSPT is also related to egocentric perspective transformations (imagined rotations or translations of one's point of view) and to infer the visuospatial experiences of another person (e.g. if and how another person sees objects). This process is deeply related to a wide-ranging capacity called the theory of mind (ToM), an essential cognitive function that allows us to regulate our social behaviour by attributing mental representations to individuals in order to make behavioural predictions. VSPT is often considered in the literature as the starting point of the development of the theory of mind. VSPT and ToM include several levels of knowledge that have to be assessed by specific tasks. Unfortunately, the lack of tasks assessing these functions in clinical neuropsychology leads to underestimate, in brain-damaged patients, deficits of these functions which are essential, in everyday life, to regulate our social behaviour (ToM) and to navigate in known and unknown environments (VSPT). Therefore, this study aims to create and standardize a VSPT task in order to explore the cognitive requirements of VSPT and ToM, and to specify their relationship in healthy adults and thereafter in brain-damaged patients. Two versions of a computerized VSPT task were administered to healthy participants (M = 28.18, SD = 4.8 years). In both versions the environment was a 3D representation of 10 different geometric shapes placed on a circular base. Two sets of eight pictures were generated from this: of the environment with an avatar somewhere on its periphery (locations) and of what the avatar sees from that place (views). Two types of questions were asked: a) identify the location from the view, and b) identify the view from the location. Twenty participants completed version 1 of the task and 20 completed the second version, where the views were offset by ±15° (i.e., clockwise or counterclockwise) and participants were asked to choose the closest location or the closest view. The preliminary findings revealed that version 1 is significantly easier than version 2 for accuracy (with ceiling scores for version 1). In version 2, participants responded significantly slower when they had to infer the avatar's view from the latter's location, probably because they spent more time visually exploring the different views (responses). Furthermore, men significantly performed better than women in version 1 but not in version 2. Most importantly, a sensitive task (version 2) has been created for which the participants do not seem to easily and automatically compute what someone is looking at yet which does not involve more heavily other cognitive functions. This study is further completed by including analysis on non-clinical participants with low and high degrees of schizotypy, different socio-educational status, and with a range of older adults to examine age-related and other differences in VSPT processing.

Keywords: mental transformation, spatial cognition, theory of mind, visuospatial perspective taking

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Authors: Alberto Mora-Vazquez, Nelly Paulina Trejo Guzmán

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This study examines the learning trajectories of twelve language teachers who were former students of a BA in applied linguistics at a Mexican state university. In particular, the study compares the social, academic and professional trajectories of two groups of teachers, six locally raised and educated ones and six repatriated ones from the U.S. Our interest in undertaking this research lies in the wide variety of students’ backgrounds we as professors in the BA program have witnessed throughout the years it has been around. Ever since the academic program started back in 2006, the student population has been made up of students whose backgrounds are highly diverse in terms of English language proficiency level, professional orientations and degree of cross-cultural awareness. Such diversity is further evidenced by the ongoing incorporation of some transnational students who have lived and studied in the United States for a significant period of time before their enrolment in the BA program. This, however, is not an isolated event as other researchers have reported this phenomenon in other TESOL-related programs of Mexican universities in the literature. Therefore, this suggests that their social and educational experiences are quite different from those of their Mexican born and educated counterparts. In addition, an informal comparison of the participation in formal teaching activities of the two groups at the beginning of their careers also suggested that significant differences in teacher training and development needs could also be identified. This issue raised questions about the need to examine the life and learning trajectories of these two groups of student teachers so as to develop an intervention plan aimed at supporting and encouraging their academic and professional advancement based on their particular needs. To achieve this goal, the study makes use of a combination of retrospective life-history research and the analysis of academic documents. The first approach uses interviews for data-collection. Through the use of a narrative life-history interview protocol, teachers were asked about their childhood home context, their language learning and teaching experiences, their stories of studying applied linguistics, and self-description. For the analysis of participants’ educational outcomes, a wide range of academic records, including reports of language proficiency exams results and language teacher training certificates, were used. The analysis revealed marked differences between the two groups of teachers in terms of academic and professional orientations. The locally educated teachers tended to graduate first, to look for further educational opportunities after graduation, to enter the language teaching profession earlier, and to expand their professional development options more than their peers. It is argued that these differences can be explained by their identities, which are made up of the interplay of influences such as their home context, their previous educational experiences and their cultural background. Implications for language teacher trainers and applied linguistics academic program administrators are provided.

Keywords: beginning language teachers, life-history research, Mexican context, transnational students

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Authors: Francisco Vidal Vázquez, Pekka Simell, Christian Frilund, Matti Reinikainen, Ilkka Hiltunen, Tim Böltken, Benjamin Andris, Paolo Piermartini

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Carbon capture and utilization (CCU) are one of the key topics in mitigation of CO2 emissions. There are many different technologies that are applied for the production of diverse chemicals from CO2 such as synthetic natural gas, Fischer-Tropsch products, methanol and polymers. Power-to-Gas and Power-to-Liquids concepts arise as a synergetic solution for storing energy and producing value added products from the intermittent renewable energy sources and CCU. VTT is a research and technology development company having energy in transition as one of the key focus areas. VTT has extensive experience in piloting and upscaling of new energy and chemical processes. Recently, VTT has developed and commissioned a Mobile Synthesis Unit (MOBSU) in close collaboration with INERATEC, a spin-off company of Karlsruhe Institute of Technology (KIT, Germany). The MOBSU is a multipurpose synthesis unit for CO2 upgrading to energy carriers and chemicals, which can be transported on-site where CO2 emission and renewable energy are available. The MOBSU is initially used for production of fuel compounds and chemical intermediates by combination of two consecutive processes: reverse Water-Gas Shift (rWGS) and Fischer-Tropsch synthesis (FT). First, CO2 is converted to CO by high-pressure rWGS and then, the CO and H2 rich effluent is used as feed for FT using an intensified reactor technology developed and designed by INERATEC. Chemical equilibrium of rWGS reaction is not affected by pressure. Nevertheless, compression would be required in between rWGS and FT in the case when rWGS is operated at atmospheric pressure. This would also require cooling of rWGS effluent, water removal and reheating. For that reason, rWGS is operated using precious metal catalyst in the MOBSU at similar pressure as FT to simplify the process. However, operating rWGS at high pressures has also some disadvantages such as methane and carbon formation, and more demanding specifications for materials. The main parts of FT module are an intensified reactor, a hot trap to condense the FT wax products, and a cold trap to condense the FT liquid products. The FT synthesis is performed using cobalt catalyst in a novel compact reactor technology with integrated highly-efficient water evaporation cooling cycle. The MOBSU started operation in November 2016. First, the FT module is tested using as feedstock H2 and CO. Subsequently, rWGS and FT modules are operated together using CO2 and H2 as feedstock of ca. 5 Nm3/hr total flowrate. On spring 2017, The MOBSU unit will be integrated together with a direct air capture (DAC) of CO2 unit, and a PEM electrolyser unit at Lappeenranta University of Technology (LUT) premises for demonstration of the SoletAir concept. This would be the first time when synthetic fuels are produced by combination of DAC unit and electrolyser unit which uses solar power for H2 production.

Keywords: CO2 utilization, demonstration, Fischer-Tropsch synthesis, intensified reactors, reverse water-gas shift

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Authors: A. F. C. Nassar, D. K. Tessler, L. Okuda, C. Del Fava, D. P. Chiebao, A. H. C. N. Romaldini, A. P. Alvim, M. J. Sanchez-Vazquez, M. S. Rosa, J. C. Pompei, R. Harakava, M. C. S. Araujo, G. H. F. Marques, E. M. Pituco

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Glanders is a zoonotic disease of Equidae caused by the bacterium Burkholderia mallei presented in acute or chronic clinical forms with inflammatory nodules in the respiratory tract, lymphangitis and caseous lymph nodes. There is not a treatment with veterinary drugs to this life-threatening disease; thus, its occurrence must be notified to official animal health services and any infected animal must be eliminated. This study aims to detect B. mallei from horses euthanized in outbreaks of glanders in Brazil, providing a better understanding of the bacterial characteristics and determine a proper protocol for isolation. The work was carried out with the collaboration of the Ministry of Agriculture and the Sao Paulo State Animal Health Department, while its procedures were approved by the Committee of Ethics in Animal Experimentation from the Instituto Biologico (CETEA n°156/2017). To the present time, 16 horses from farms with outbreaks of glanders detected by complement fixation test (CFT) serology method were analyzed. During the necropsy, samples of possibly affected organs (lymph nodes, lungs, heart, liver, spleen, kidneys and trachea) were collected for bacterial isolation, molecular tests and pathology. Isolation was performed using two enriched mediums, a potato infusion agar with 5% sheep blood, 4% glycerol and antibiotics (penicilin100U/ mL), and another with the same ingredients except the antibiotic. A PCR protocol was modified for this study using primers design to identify a region of the Flip gen of B. mallei. Thru isolation, 12.5% (2/16) animals were confirmed positive using only the enriched medium with antibiotic and confirmed by PCR: from mediastinal and submandibular lymph nodes and lungs in one animal and from mediastinal lymph node in the other. The detection of the bacterium using PCR showed positivity of 100% (16/16) horses from 144 samples of organs. Pathology macroscopic lesions observed were catarrhal nasal discharge, fetlock ulcers, emaciation, lymphangitis in limbs, suppurative lymphangitis, lymph node enlargement, star shaped liver, and spleen scars, adherence of the renal capsule, pulmonary hemorrhage, and miliary nodules. Microscopic lesions were suppurative bronchopneumonia with microabscesses and Langhans giant cells in lungs; lymph nodes with abscesses and intense lymphoid reaction; hemosiderosis and abscesses in spleen. Positive samples on PCR will be sequenced later and analyzed comparing with previous records in the literature. A throughout description of the recent acute cases of glanders occurring in Brazil and characterization of the bacterium related will contribute to advances in the knowledge of the pathogenicity, clinical symptoms, and epidemiology of this zoonotic disease. Acknowledgment: This project is sponsored by FAPESP.

Keywords: equines, bacterial isolation, zoonosis, PCR, pathology

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Authors: Andres Nieto-Leal, Victor N. Kaliakin, Tania P. Molina

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The nature of most engineering materials is quite complex. It is, therefore, difficult to devise a general mathematical model that will cover all possible ranges and types of excitation and behavior of a given material. As a result, the development of mathematical models is based upon simplifying assumptions regarding material behavior. Such simplifications result in some material idealization; for example, one of the simplest material idealization is to assume that the material behavior obeys the elasticity. However, soils are nonhomogeneous, anisotropic, path-dependent materials that exhibit nonlinear stress-strain relationships, changes in volume under shear, dilatancy, as well as time-, rate- and temperature-dependent behavior. Over the years, many constitutive models, possessing different levels of sophistication, have been developed to simulate the behavior geomaterials, particularly cohesive soils. Early in the development of constitutive models, it became evident that elastic or standard elastoplastic formulations, employing purely isotropic hardening and predicated in the existence of a yield surface surrounding a purely elastic domain, were incapable of realistically simulating the behavior of geomaterials. Accordingly, more sophisticated constitutive models have been developed; for example, the bounding surface elastoplasticity. The essence of the bounding surface concept is the hypothesis that plastic deformations can occur for stress states either within or on the bounding surface. Thus, unlike classical yield surface elastoplasticity, the plastic states are not restricted only to those lying on a surface. Elastoplastic bounding surface models have been improved; however, there is still need to improve their capabilities in simulating the response of anisotropically consolidated cohesive soils, especially the response in extension tests. Thus, in this work an improved constitutive model that can more accurately predict diverse stress-strain phenomena exhibited by cohesive soils was developed. Particularly, an improved rotational hardening rule that better simulate the response of cohesive soils in extension. The generalized definition of the bounding surface model provides a convenient and elegant framework for unifying various previous versions of the model for anisotropically consolidated cohesive soils. The Generalized Bounding Surface Model for cohesive soils is a fully three-dimensional, time-dependent model that accounts for both inherent and stress induced anisotropy employing a non-associative flow rule. The model numerical implementation in a computer code followed an adaptive multistep integration scheme in conjunction with local iteration and radial return. The one-step trapezoidal rule was used to get the stiffness matrix that defines the relationship between the stress increment and the strain increment. After testing the model in simulating the response of cohesive soils through extensive comparisons of model simulations to experimental data, it has been shown to give quite good simulations. The new model successfully simulates the response of different cohesive soils; for example, Cardiff Kaolin, Spestone Kaolin, and Lower Cromer Till. The simulated undrained stress paths, stress-strain response, and excess pore pressures are in very good agreement with the experimental values, especially in extension.

Keywords: bounding surface elastoplasticity, cohesive soils, constitutive model, modeling of geomaterials

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Authors: Beste Ozcan, Valerio Sperati, Laura Romano, Tania Moretta, Simone Scaffaro, Noemi Faedda, Federica Giovannone, Carla Sogos, Vincenzo Guidetti, Gianluca Baldassarre

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+me is an experimental interactive toy with the appearance of a soft, pillow-like, panda. Shape and consistency are designed to arise emotional attachment in young children: a child can wear it around his/her neck and treat it as a companion (i.e. a transitional object). When caressed on paws or head, the panda emits appealing, interesting outputs like colored lights or amusing sounds, thanks to embedded electronics. Such sensory patterns can be modified through a wirelessly connected tablet: by this, an adult caregiver can adapt +me responses to a child's reactions or requests, for example, changing the light hue or the type of sound. The toy control is therefore shared, as it depends on both the child (who handles the panda) and the adult (who manages the tablet and mediates the sensory input-output contingencies). These features make +me a potential tool for therapy with children with Neurodevelopmental Disorders (ND), characterized by impairments in the social area, like Autism Spectrum Disorders (ASD) and Language Disorders (LD): as a proposal, the toy could be used together with a therapist, in rehabilitative play activities aimed at encouraging simple social interactions and reinforcing basic relational and communication skills. +me was tested in two pilot experiments, the first one involving 15 Typically Developed (TD) children aged in 8-34 months, the second one involving 7 children with ASD, and 7 with LD, aged in 30-48 months. In both studies a researcher/caregiver, during a one-to-one, ten-minute activity plays with the panda and encourages the child to do the same. The purpose of both studies was to ascertain the general acceptability of the device as an interesting toy that is an object able to capture the child's attention and to maintain a high motivation to interact with it and with the adult. Behavioral indexes for estimating the interplay between the child, +me and caregiver were rated from the video recording of the experimental sessions. Preliminary results show how -on average- participants from 3 groups exhibit a good engagement: they touch, caress, explore the panda and show enjoyment when they manage to trigger luminous and sound responses. During the experiments, children tend to imitate the caregiver's actions on +me, often looking (and smiling) at him/her. Interesting behavioral differences between TD, ASD, and LD groups are scored: for example, ASD participants produce a fewer number of smiles both to panda and to a caregiver with respect to TD group, while LD scores stand between ASD and TD subjects. These preliminary observations suggest that the interactive toy +me is able to raise and maintain the interest of toddlers and therefore it can be reasonably used as a supporting tool during therapy, to stimulate pivotal social skills as imitation, turn-taking, eye contact, and social smiles. Interestingly, the young age of participants, along with the behavioral differences between groups, seem to suggest a further potential use of the device: a tool for early differential diagnosis (the average age of a child

Keywords: autism spectrum disorders, interactive toy, social interaction, therapy, transitional wearable companion

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Authors: Velitchko G. Tzatchkov, Petronilo E. Cortes-Mejia, J. Manuel Rodriguez-Varela, Jesus Figueroa-Vazquez

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Hydraulic headloss, expressed by the values of friction factor f and Manning’s coefficient n, is an important parameter in designing drainage pipes. Their values normally are taken from manufacturer recommendations, many times without sufficient experimental support. To our knowledge, currently there is no standard procedure for hydraulically testing such pipes. As a result of research carried out at the Mexican Institute of Water Technology, a laboratory testing procedure was proposed and applied on 6 and 12 inches diameter polyvinyl chloride (PVC) and high-density dual wall polyethylene pipe (HDPE) drainage pipes. While the PVC pipe is characterized by naturally smooth interior and exterior walls, the dual wall HDPE pipe has corrugated exterior wall and, although considered smooth, a slightly wavy interior wall. The pipes were tested at full and partially full pipe flow conditions. The tests for full pipe flow were carried out on a 31.47 m long pipe at flow velocities between 0.11 and 4.61 m/s. Water was supplied by gravity from a 10 m-high tank in some of the tests, and from a 3.20 m-high tank in the rest of the tests. Pressure was measured independently with piezometer readings and pressure transducers. The flow rate was measured by an ultrasonic meter. For the partially full pipe flow the pipe was placed inside an existing 49.63 m long zero slope (horizontal) channel. The flow depth was measured by piezometers located along the pipe, for flow rates between 2.84 and 35.65 L/s, measured by a rectangular weir. The observed flow profiles were then compared to computer generated theoretical gradually varied flow profiles for different Manning’s n values. It was found that Manning’s n, that normally is assumed constant for a given pipe material, is in fact dependent on flow velocity and pipe diameter for full pipe flow, and on flow depth for partially full pipe flow. Contrary to the expected higher values of n and f for the HDPE pipe, virtually the same values were obtained for the smooth interior wall PVC pipe and the slightly wavy interior wall HDPE pipe. The explanation of this fact was found in Henry Morris’ theory for smooth turbulent conduit flow over isolated roughness elements. Following Morris, three categories of the flow regimes are possible in a rough conduit: isolated roughness (or semi smooth turbulent) flow, wake interference (or hyper turbulent) flow, and skimming (or quasi-smooth) flow. Isolated roughness flow is characterized by friction drag turbulence over the wall between the roughness elements, independent vortex generation, and dissipation around each roughness element. In this regime, the wake and vortex generation zones at each element develop and dissipate before attaining the next element. The longitudinal spacing of the roughness elements and their height are important influencing agents. Given the slightly wavy form of the HDPE pipe interior wall, the flow for this type of pipe belongs to this category. Based on that theory, an equation for the hydraulic friction factor was obtained. The obtained coefficient values are going to be used in the Mexican design standards.

Keywords: drainage plastic pipes, hydraulic headloss, hydraulic friction factor, Manning’s n

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Authors: Iñaki Iturria, Pasquale Russo, Montserrat Nacher-Vázquez, Giuseppe Spano, Paloma López, Miguel Angel Pardo

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Introduction: It is known that some Lactic Acid Bacteria (LAB) present an interesting probiotic effect. Probiotic bacteria stimulate host resistance to microbial pathogens and thereby aid in immune response, and modulate the host's immune responses to antigens with a potential to down-regulate hypersensitivity reactions. Therefore, probiotic therapy is valuable against intestinal infections and may be beneficial in the treatment of Inflammatory Bowel Disease (IBD). Several in vitro tests are available to evaluate the probiotic potential of a LAB strain. However, an in vivo model is required to understand the interaction between the host immune system and the bacteria. During the last few years, zebrafish (Danio rerio) has gained interest as a promising vertebrate model in this field. This organism has been extensively used to study the interaction between the host and the microbiota, as well as the host immune response under several microbial infections. In this work, we report on the use of the zebrafish model to investigate in vivo the colonizing ability and the immunomodulatory effect of probiotic LAB. Methods: Lactobacillus strains belonging to different LAB species were fluorescently tagged and used to colonize germ-free zebrafish larvae gastrointestinal tract (GIT). Some of the strains had a well-documented probiotic effect (L. acidophilus LA5); while others presented an exopolysaccharide (EPS) producing phenotype, thus allowing evaluating the influence of EPS in the colonization and immunomodulatory effect. Bacteria colonization was monitored for 72 h by direct observation in real time using fluorescent microscopy. CFU count per larva was also evaluated at different times. The immunomodulatory effect was assessed analysing the differential expression of several innate immune system genes (MyD88, NF-κB, Tlr4, Il1β and Il10) by qRT- PCR. The anti-inflammatory effect was evaluated using a chemical enterocolitis zebrafish model. The protective effect against a pathogen was also studied. To that end, a challenge test was developed using a fluorescently tagged pathogen (Vibrio anguillarum-GFP+). The progression of the infection was monitored up to 3 days using a fluorescent stereomicroscope. Mortality rates and CFU counts were also registered. Results and conclusions: Larvae exposed to EPS-producing bacteria showed a higher fluorescence and CFU count than those colonized with no-EPS phenotype LAB. In the same way, qRT-PCR results revealed an immunomodulatory effect on the host after the administration of the strains with probiotic activity. A downregulation of proinflammatory cytoquines as well as other cellular mediators of inflammation was observed. The anti-inflammatory effect was found to be particularly marked following exposure to LA% strain, as well as EPS producing strains. Furthermore, the challenge test revealed a protective effect of probiotic administration. As a matter of fact, larvae fed with probiotics showed a decrease in the mortality rate ranging from 20 to 35%. Discussion: In this work, we developed a promising model, based on the use of gnotobiotic zebrafish coupled with a bacterial fluorescent tagging in order to evaluate the probiotic potential of different LAB strains. We have successfully used this system to monitor in real time the colonization and persistence of exogenous LAB within the gut of zebrafish larvae, to evaluate their immunomodulatory effect and for in vivo competition assays. This approach could bring further insights into the complex microbial-host interactions at intestinal level.

Keywords: gnotobiotic, immune system, lactic acid bacteria, probiotics, zebrafish

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Authors: Diana Santiago, Tania Lisboa, Sophie Lee, Alexander P. Demos, Monica C. S. Vasconcelos

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Memorization has long been recognized as an arduous and anxiety-evoking task for musicians, and yet, it is an essential aspect of performance. Research shows that musicians are often not taught how to memorize. While memorization and practice strategies of professionals have been studied, little research has been done to examine how student musicians learn to practice and memorize music in different cultural settings. We present the process of developing and testing a questionnaire of music memorization and musical practice for student musicians in the UK and Brazil. A survey was developed for a cross-cultural research project aiming at examining how young orchestral musicians (aged 7–18 years) in different learning environments and cultures engage in instrumental practice and memorization. The questionnaire development included members of a UK/US/Brazil research team of music educators and performance science researchers. A pool of items was developed for each aspect of practice and memorization identified, based on literature, personal experiences, and adapted from existing questionnaires. Item development took the varying levels of cognitive and social development of the target populations into consideration. It also considered the diverse target learning environments. Items were initially grouped in accordance with a single underlying construct/behavior. The questionnaire comprised three sections: a demographics section, a section on practice (containing 29 items), and a section on memorization (containing 40 items). Next, the response process was considered and a 5-point Likert scale ranging from ‘always’ to ‘never’ with a verbal label and an image assigned to each response option was selected, following effective questionnaire design for children and youths. Finally, a pilot study was conducted with young orchestral musicians from diverse learning environments in Brazil and the United Kingdom. Data collection took place in either one-to-one or group settings to facilitate the participants. Cognitive interviews were utilized to establish response process validity by confirming the readability and accurate comprehension of the questionnaire items or highlighting the need for item revision. Internal reliability was investigated by measuring the consistency of the item groups using the statistical test Cronbach’s alpha. The pilot study successfully relied on the questionnaire to generate data about the engagement of young musicians of different levels and instruments, across different learning and cultural environments, in instrumental practice and memorization. Interaction analysis of the cognitive interviews undertaken with these participants, however, exposed the fact that certain items, and the response scale, could be interpreted in multiple ways. The questionnaire text was, therefore, revised accordingly. The low Cronbach’s Alpha scores of many item groups indicated another issue with the original questionnaire: its low level of internal reliability. Several reasons for each poor reliability can be suggested, including the issues with item interpretation revealed through interaction analysis of the cognitive interviews, the small number of participants (34), and the elusive nature of the construct in question. The revised questionnaire measures 78 specific behaviors or opinions. It can be seen to provide an efficient means of gathering information about the engagement of young musicians in practice and memorization on a large scale.

Keywords: cross-cultural, memorization, practice, questionnaire, young musicians

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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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