Search results for: Paula Ferreira
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 303

Search results for: Paula Ferreira

63 Generation Z: Insights into Travel Behavior

Authors: Joao Ferreira Do Rosario, Nuno Gustavo, Ana Machado, Lurdes Calisto, Luisa Carvalho, Georgette Andraz

Abstract:

Currently, tourism small and medium enterprises (TSMEs) face serious economic and financial problems, making recovery efforts difficult. How the pandemic will affect tourists' behavior is still to be known. Will tourists be even more cautious regarding their choices or, on the contrary, will they be more adventurers with an enormous desire to travel in search of the lost freedom? Tourists may become even more demanding when traveling, more austere, or less concerned and eager to socialize. Adjusting to this "new tourist" is an added challenge for tourism service providers. Generation Z made up of individuals born in 1995 and following years, currently tends to assume a particular role and meaning in the present and future economic and social context, considering that we are facing the youngest workforce as well as tomorrow's consumers. This generation is distinguished from others as it is the first generation to combine a high level of education and technological knowledge and to fully experience the digital world. These young people are framed by a new value system that can explain new behaviours and consumption, namely, in the context of tourism. All these considerations point to the importance of investigating this target group as it is essential to understand how these individuals perceive, understand, act, and can be involved in a new environment built around a society regulated by new priorities and challenges of a sustainable nature. This leads not only to a focus on short-term market choices but mainly to predict future choices from a longer-term perspective. Together with the social background of a person, values are considered a stable antecedent of behavior and might therefore predict not just immediate, but also future choices. Furthermore, the meaning attributed to travel has a general connotation and goes beyond a specific travel choice or experience. In other words, values and travel's meaning form a chain of influences on the present and future travel behavior. This study explores the social background and values of Generation Z travelers vs the meaning these tourists give to travel. The aim is to discover in their present behavior cues to predict travel choices so that the future of tourism can be secured. This study also provides data for predicting the tourism choices of youngsters in the more immediate future. Methodologically, a quantitative approach was adopted based on the collection of data through a survey. Since academic research on Generation Z of tourists is still scarce, it is expected to contribute to deepening scientific knowledge in this area. Furthermore, it is expected that this research will support tourism professionals in defining differentiated marketing strategies and adapted to the requirements of this target, in a new time.

Keywords: Generation Z, travel behavior, travel meaning, Generation Z Values

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62 Antimicrobial Properties of SEBS Compounds with Copper Microparticles

Authors: Vanda Ferreira Ribeiro, Daiane Tomacheski, Douglas Naue Simões, Michele Pitto, Ruth Marlene Campomanes Santana

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Indoor environments, such as car cabins and public transportation vehicles are places where users are subject to air quality. Microorganisms (bacteria, fungi, yeasts) enter these environments through windows, ventilation systems and may use the organic particles present as a growth substrate. In addition, atmospheric pollutants can act as potential carbon and nitrogen sources for some microorganisms. Compounds base SEBS copolymers, poly(styrene-b-(ethylene-co-butylene)-b-styrene, are a class of thermoplastic elastomers (TPEs), fully recyclable and largely used in automotive parts. Metals, such as cooper and silver, have biocidal activities and the production of the SEBS compounds by melting blending with these agents can be a good option for producing compounds for use in plastic parts of ventilation systems and automotive air-conditioning, in order to minimize the problems caused by growth of pathogenic microorganisms. In this sense, the aim of this work was to evaluate the effect of copper microparticles as antimicrobial agent in compositions based on SEBS/PP/oil/calcite. Copper microparticles were used in weight proportion of 0%, 1%, 2% and 4%. The compounds were prepared using a co-rotating double screw extruder (L/D ratio of 40/1 and 16 mm screw diameter). The processing parameters were 300 rpm of screw rotation rate, with a temperature profile between 150 to 190°C. SEBS based TPE compounds were injection molded. The compounds emission were characterized by gravimetric fogging test. Compounds were characterized by physical (density and staining by contact), mechanical (hardness and tension properties) and rheological properties (melt volume rate – MVR). Antibacterial properties were evaluated against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) strains. To avaluate the abilities toward the fungi have been chosen Aspergillus niger (A. niger), Candida albicans (C. albicans), Cladosporium cladosporioides (C. cladosporioides) and Penicillium chrysogenum (P. chrysogenum). The results of biological tests showed a reduction on bacteria in up to 88% in E.coli and up to 93% in S. aureus. The tests with fungi showed no conclusive results because the sample without copper also demonstrated inhibition of the development of these microorganisms. The copper addition did not cause significant variations in mechanical properties, in the MVR and the emission behavior of the compounds. The density increases with the increment of copper in compounds.

Keywords: air conditioner, antimicrobial, cooper, SEBS

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61 Change of Substrate in Solid State Fermentation Can Produce Proteases and Phytases with Extremely Distinct Biochemical Characteristics and Promising Applications for Animal Nutrition

Authors: Paula K. Novelli, Margarida M. Barros, Luciana F. Flueri

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Utilization of agricultural by-products, wheat ban and soybean bran, as substrate for solid state fermentation (SSF) was studied, aiming the achievement of different enzymes from Aspergillus sp. with distinct biological characteristics and its application and improvement on animal nutrition. Aspergillus niger and Aspergillus oryzea were studied as they showed very high yield of phytase and protease production, respectively. Phytase activity was measure using p-nitrophenilphosphate as substrate and a standard curve of p-nitrophenol, as the enzymatic activity unit was the quantity of enzyme necessary to release one μmol of p-nitrophenol. Protease activity was measure using azocasein as substrate. Activity for phytase and protease substantially increased when the different biochemical characteristics were considered in the study. Optimum pH and stability of the phytase produced by A. niger with wheat bran as substrate was between 4.0 - 5.0 and optimum temperature of activity was 37oC. Phytase fermented in soybean bran showed constant values at all pHs studied, for optimal and stability, but low production. Phytase with both substrates showed stable activity for temperatures higher than 80oC. Protease from A. niger showed very distinct behavior of optimum pH, acid for wheat bran and basic for soybean bran, respectively and optimal values of temperature and stability at 50oC. Phytase produced by A. oryzae in wheat bran had optimum pH and temperature of 9 and 37oC, respectively, but it was very unstable. On the other hand, proteases were stable at high temperatures, all pH’s studied and showed very high yield when fermented in wheat bran, however when it was fermented in soybean bran the production was very low. Subsequently the upscale production of phytase from A. niger and proteases from A. oryzae were applied as an enzyme additive in fish fed for digestibility studies. Phytases and proteases were produced with stable enzyme activity of 7,000 U.g-1 and 2,500 U.g-1, respectively. When those enzymes were applied in a plant protein based fish diet for digestibility studies, they increased protein, mineral, energy and lipids availability, showing that these new enzymes can improve animal production and performance. In conclusion, the substrate, as well as, the microorganism species can affect the biochemical character of the enzyme produced. Moreover, the production of these enzymes by SSF can be up to 90% cheaper than commercial ones produced with the same fungi species but submerged fermentation. Add to that these cheap enzymes can be easily applied as animal diet additives to improve production and performance.

Keywords: agricultural by-products, animal nutrition, enzymes production, solid state fermentation

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60 Benefits of Tourist Experiences for Families: A Systematic Literature Review Using Nvivo

Authors: Diana Cunha, Catarina Coelho, Ana Paula Relvas, Elisabeth Kastenholz

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Context: Tourist experiences have a recognized impact on the well-being of individuals. However, studies on the specific benefits of tourist experiences for families are scattered across different disciplines. This study aims to systematically review the literature to synthesize the evidence on the benefits of tourist experiences for families. Research Aim: The main objective is to systematize the evidence in the literature regarding the benefits of tourist experiences for families. Methodology: A systematic literature review was conducted using Nvivo, analyzing 33 scientific studies obtained from various databases. The search terms used were "family"/ "couple" and "tourist experience". The studies included quantitative, qualitative, mixed methods, and literature reviews. All works prior to the year 2000 were excluded, and the search was restricted to full text. A language filter was also used, considering articles in Portuguese, English, and Spanish. For NVivo analysis, information was coded based on both deductive and inductive perspectives. To minimize the subjectivity of the selection and coding process, two of the authors discussed the process and agreed on criteria that would make the coding more objective. Once the coding process in NVivo was completed, the data relating to the identification/characterization of the works were exported to the Statistical Package for the Social Sciences (SPPS), to characterize the sample. Findings: The results highlight that tourist experiences have several benefits for family systems, including the strengthening of family and marital bonds, the creation of family memories, and overall well-being and life satisfaction. These benefits contribute to both immediate relationship quality improvement and long-term family identity construction and transgenerational transmission. Theoretical Importance: This study emphasizes the systemic nature of the effects and relationships within family systems. It also shows that no harm was reported within these experiences, with only some challenges related to positive outcomes. Data Collection and Analysis Procedures: The study collected data from 33 scientific studies published predominantly after 2013. The data were analyzed using Nvivo, employing a systematic review approach. Question Addressed: The study addresses the question of the benefits of tourist experiences for families and how these experiences contribute to family functioning and individual well-being. Conclusion: Tourist experiences provide opportunities for families to enhance their interpersonal relationships and create lasting memories. The findings suggest that formal interventions based on evidence could further enhance the potential benefits of these experiences and be a valuable preventive tool in therapeutic interventions.

Keywords: family systems, individual and family well-being, marital satisfaction, tourist experiences

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59 The Influence of Hydrolyzed Cartilage Collagen on General Mobility and Wellbeing of an Active Population

Authors: Sara De Pelsmaeker, Catarina Ferreira da Silva, Janne Prawit

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Recent studies show that enzymatically hydrolysed collagen is absorbed and distributed to joint tissues, where it has analgesic and active anti-inflammatory properties. Reviews of the associated relevant literature also support this theory. However, these studies are all using hydrolyzed collagen from animal hide or skin. This study looks into the effect of daily supplementation of hydrolyzed cartilage collagen (HCC), which has a different composition. A consumer study was set up using a double-blind placebo-controlled design with a control group using twice a day 0.5gr of maltodextrin and an experimental group using twice 0.5g of HCC, over a trial period of 12 weeks. A follow-up phase of 4 weeks without supplementation was taken into the experiment to investigate the ‘wash-out’ phase. As this consumer study was conducted during the lockdown periods, a specific app was designed to follow up with the participants. The app had the advantage that in this way, the motivation of the participants was enhanced and the drop-out range of participants was lower than normally seen in consumer studies. Participants were recruited via various sports and health clubs across the UK as we targeted a general population of people that considered themselves in good health. Exclusion criteria were ‘not experiencing any medical conditions’ and ‘not taking any prescribed medication’. A minimum requirement was that they regularly engaged in some level of physical activity. The participants had to log the type of activity that they conducted and the duration of the activity. Weekly, participants were providing feedback on their joint health and subjective pain using the validated pain measuring instrument Visual Analogue Scale (VAS). The weekly repoAbstract Public Health and Wellbeing Conferencerting section in the app was designed with simplicity and based on the accuracy demonstrated in previous similar studies to track subjective pain measures of participants. At the beginning of the trial, each participant indicated their baseline on joint pain. The results of this consumer study indicated that HCC significantly improved joint health and subjective pain scores compared to the placebo group. No significant differences were found between different demographic groups (age or gender). The level of activity, going from high intensive training to regular walking, did not significantly influence the effect of the HCC. The results of the wash-out phase indicated that when the participants stopped the HCC supplementation, their subjective pain scores increased again to the baseline. In conclusion, the results gave a positive indication that the daily supplementation of HCC can contribute to the overall mobility and wellbeing of a general active population

Keywords: VAS-score, food supplement, mobility, joint health

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58 Structural and Biochemical Characterization of Red and Green Emitting Luciferase Enzymes

Authors: Wael M. Rabeh, Cesar Carrasco-Lopez, Juliana C. Ferreira, Pance Naumov

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Bioluminescence, the emission of light from a biological process, is found in various living organisms including bacteria, fireflies, beetles, fungus and different marine organisms. Luciferase is an enzyme that catalyzes a two steps oxidation of luciferin in the presence of Mg2+ and ATP to produce oxyluciferin and releases energy in the form of light. The luciferase assay is used in biological research and clinical applications for in vivo imaging, cell proliferation, and protein folding and secretion analysis. The luciferase enzyme consists of two domains, a large N-terminal domain (1-436 residues) that is connected to a small C-terminal domain (440-544) by a flexible loop that functions as a hinge for opening and closing the active site. The two domains are separated by a large cleft housing the active site that closes after binding the substrates, luciferin and ATP. Even though all insect luciferases catalyze the same chemical reaction and share 50% to 90% sequence homology and high structural similarity, they emit light of different colors from green at 560nm to red at 640 nm. Currently, the majority of the structural and biochemical studies have been conducted on green-emitting firefly luciferases. To address the color emission mechanism, we expressed and purified two luciferase enzymes with blue-shifted green and red emission from indigenous Brazilian species Amydetes fanestratus and Phrixothrix, respectively. The two enzymes naturally emit light of different colors and they are an excellent system to study the color-emission mechanism of luciferases, as the current proposed mechanisms are based on mutagenesis studies. Using a vapor-diffusion method and a high-throughput approach, we crystallized and solved the crystal structure of both enzymes, at 1.7 Å and 3.1 Å resolution respectively, using X-ray crystallography. The free enzyme adopted two open conformations in the crystallographic unit cell that are different from the previously characterized firefly luciferase. The blue-shifted green luciferase crystalized as a monomer similar to other luciferases reported in literature, while the red luciferases crystalized as an octamer and was also purified as an octomer in solution. The octomer conformation is the first of its kind for any insect’s luciferase, which might be relate to the red color emission. Structurally designed mutations confirmed the importance of the transition between the open and close conformations in the fine-tuning of the color and the characterization of other interesting mutants is underway.

Keywords: bioluminescence, enzymology, structural biology, x-ray crystallography

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57 Thermoregulatory Responses of Holstein Cows Exposed to Intense Heat Stress

Authors: Rodrigo De A. Ferrazza, Henry D. M. Garcia, Viviana H. V. Aristizabal, Camilla De S. Nogueira, Cecilia J. Verissimo, Jose Roberto Sartori, Roberto Sartori, Joao Carlos P. Ferreira

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Environmental factors adversely influence sustainability in livestock production system. Dairy herds are the most affected by heat stress among livestock industries. This clearly implies in development of new strategies for mitigating heat, which should be based on physiological and metabolic adaptations of the animal. In this study, we incorporated the effect of climate variables and heat exposure time on the thermoregulatory responses in order to clarify the adaptive mechanisms for bovine heat dissipation under intense thermal stress induced experimentally in climate chamber. Non-lactating Holstein cows were contemporaneously and randomly assigned to thermoneutral (TN; n=12) or heat stress (HS; n=12) treatments during 16 days. Vaginal temperature (VT) was measured every 15 min with a microprocessor-controlled data logger (HOBO®, Onset Computer Corporation, Bourne, MA, USA) attached to a modified vaginal controlled internal drug release insert (Sincrogest®, Ourofino, Brazil). Rectal temperature (RT), respiratory rate (RR) and heart rate (HR) were measured twice a day (0700 and 1500h) and dry matter intake (DMI) was estimated daily. The ambient temperature and air relative humidity were 25.9±0.2°C and 73.0±0.8%, respectively for TN, and 36.3± 0.3°C and 60.9±0.9%, respectively for HS. Respiratory rate of HS cows increased immediately after exposure to heat and was higher (76.02±1.70bpm; P<0.001) than TN (39.70±0.71bpm), followed by rising of RT (39.87°C±0.07 for HS versus 38.56±0.03°C for TN; P<0.001) and VT (39.82±0.10°C for HS versus 38.26±0.03°C for TN; P<0.001). A diurnal pattern was detected, with higher (P<0.01) afternoon temperatures than morning and this effect was aggravated for HS cows. There was decrease (P<0.05) of HR for HS cows (62.13±0.99bpm) compared to TN (66.23±0.79bpm), but the magnitude of the differences was not the same over time. From the third day, there was a decrease of DMI for HS in attempt to maintain homeothermy, while TN cows increased DMI (8.27kg±0.33kg d-1 for HS versus 14.03±0.29kg d-1 for TN; P<0.001). By regression analysis, RT and RR better reflected the response of cows to changes in the Temperature Humidity Index and the effect of climate variables from the previous day to influence the physiological parameters and DMI was more important than the current day, with ambient temperature the most important factor. Comparison between acute (0 to 3 days) and chronic (13 to 16 days) exposure to heat stress showed decreasing of the slope of the regression equations for RR and DMI, suggesting an adaptive adjustment, however with no change for RT. In conclusion, intense heat stress exerted strong influence on the thermoregulatory mechanisms, but the acclimation process was only partial.

Keywords: acclimation, bovine, climate chamber, hyperthermia, thermoregulation

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56 The Role of Self-Compassion for the Diagnosis of Social Anxiety Disorder in Adolescents

Authors: Diana Vieira Figueiredo, Rita Ramos Miguel, Maria do Céu Salvador, Luiza Nobre-Lima, Daniel RIjo, Paula Vagos

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Social Anxiety Disorder (SAD) is characterized by a marked and persistent fear of social and/or performance situations in which one may be exposed to the scrutiny of others.  SAD has its usual onset and is highly prevalent during adolescence; if left untreated, it often has a chronic and unremitting course. So, it seems important to understand the psychological processes that might predict the development of SAD. One of these processes may be self-compassion, which has been found to be associated with social anxiety in both adults and adolescents. Self-compassion involves three main components, each with a positive (compassionate behavior) and negative (uncompassionate behavior) pole – self-kindness versus self-judgment, common humanity versus isolation, and mindfulness versus over-identification. The negative indicators of self-compassion (self-judgement, isolation, and over-identification) were found to be more strongly linked to mental health problems than the positive indicators (self-kindness, common humanity, and mindfulness). Additionally, negative associations were found between the positive indicators of self-compassion (self-kindness, common humanity, mindfulness) and psychopathology. The current study aimed to investigate the role of self-kindness, self-judgment, common humanity, isolation, mindfulness, and over-identification in the likelihood of an adolescent presenting SAD by comparing groups of normative and socially anxious adolescents. The sample consisted of 32 adolescents (Mage = 15.88, SD = .833) of which 23 were girls. Adolescents were assessed through a clinical structured interview that led 17 to be assigned to the clinical group (presenting a primary diagnosis of SAD) and 15 to be assigned to the non-clinical group (presenting no clinical diagnosis). Variables under study were measured through the Self-Compassion Scale for adolescents (SCS-A), which assesses the six indicators of self-compassion presented above. Six separate models were tested, each with one of the subscales of the SCS-A as the independent variable and with the group (clinical versus non-clinical) as the dependent variable. The models considering isolation, over-identification, self-judgement, and self-kindness fitted the data and accurately predicted group belonging for between 75% to 84.4% of cases. Results indicated that the log of the odds of an adolescent presenting SAD was positively related to isolation, over-identification, and self-judgement and negatively associated with self-kindness. Findings provide support for the idea that decreased self-compassion may place adolescents at increased risk for experiencing clinical levels of social anxiety: on the one hand, adolescents with higher levels of isolation, over-identification, and self-judgement seem to be more prone to the development of psychopathological levels of social anxiety; on the other hand, self-kindness may play a protective role in the development of SAD in this developmental phase. So, if focusing on social feared consequences and perceiving to be different from others may be distinctive features of SAD, developing self-kindness may be the antidote to promote diminished levels of social anxiety and more.

Keywords: adolescents, social anxiety disorder, self-compassion, diagnosis odds-ration

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55 A Mixed-Method Study Exploring Expressive Writing as a Brief Intervention Targeting Mental Health and Wellbeing in Higher Education Students: A Focus on the Qualitative Findings

Authors: Deborah Bailey-Rodriguez, Maria Paula Valdivieso Rueda, Gemma Reynolds

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In recent years, the mental health of Higher Education (HE) students has been a growing concern. This has been further exacerbated by the stresses associated with the Covid-19 pandemic, placing students at even greater risk of developing mental health issues. Support available to students in HE tends to follow an established and traditional route. The demands for counseling services have grown, not only with the increase in student numbers but with the number of students seeking support for mental health issues, with 94% of HE institutions recently reporting an increase in the need for counseling services. One way of improving the well-being and mental health of HE students is through the use of brief interventions, such as expressive writing (EW). This intervention involves encouraging individuals to write continuously for at least 15-20 minutes for three to five sessions (often on consecutive days) about their deepest thoughts and feelings to explore significant personal experiences in a meaningful way. Given the brevity, simplicity and cost-effectiveness of EW, this intervention has considerable potential as an intervention for HE populations. The current study, therefore, employed a mixed-methods design to explore the effectiveness of EW in reducing anxiety, general stress, academic stress and depression in HE students while improving well-being. HE students at MDX were randomly assigned to one of three conditions: (1) The UniExp-EW group was required to write about their emotions and thoughts about any stressors they have faced that are directly relevant to their university experience (2) The NonUniExp-EW group was required to write about their emotions and thoughts about any stressors that are NOT directly relevant to their university experience, and (3) The Control group were required to write about how they spent their weekend, with no reference to thoughts or emotions, and without thinking about university. Participants were required to carry out the EW intervention for 15 minutes per day for four consecutive days. Baseline mental health and well-being measures were taken before the intervention via a battery of standardized questionnaires. Following completion of the intervention on day four, participants were required to complete the questionnaires a second time and again one week later. Participants were also invited to attend focus groups to discuss their experience of the intervention. This will allow an in-depth investigation into students’ perceptions of EW as an effective intervention to determine whether they would choose to use this intervention in the future. Preliminary findings will be discussed at the conference as well as a discussion of the important implications of the findings. The study is fundamental because if EW is an effective intervention for improving mental health and well-being in HE students, its brevity and simplicity mean it can be easily implemented and can be freely available to students. Improving the mental health and well-being of HE students can have knock-on implications for improving academic skills and career development.

Keywords: expressive writing, higher education, psychology in education, mixed-methods, mental health, academic stress

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54 A Clinical Cutoff to Identify Metabolically Unhealthy Obese and Normal-Weight Phenotype in Young Adults

Authors: Lívia Pinheiro Carvalho, Luciana Di Thommazo-Luporini, Rafael Luís Luporini, José Carlos Bonjorno Junior, Renata Pedrolongo Basso Vanelli, Manoel Carneiro de Oliveira Junior, Rodolfo de Paula Vieira, Renata Trimer, Renata G. Mendes, Mylène Aubertin-Leheudre, Audrey Borghi-Silva

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Rationale: Cardiorespiratory fitness (CRF) and functional capacity in young obese and normal-weight people are associated with metabolic and cardiovascular diseases and mortality. However, it remains unclear whether their metabolically healthy (MH) or at risk (AR) phenotype influences cardiorespiratory fitness in this vulnerable population such as obese adults but also in normal-weight people. HOMA insulin resistance index (HI) and leptin-adiponectin ratio (LA) are strong markers for characterizing those phenotypes that we hypothesized to be associated with physical fitness. We also hypothesized that an easy and feasible exercise test could identify a subpopulation at risk to develop metabolic and related disorders. Methods: Thirty-nine sedentary men and women (20-45y; 18.530 kg.m-2) underwent a clinical evaluation, including the six-minute step test (ST), a well-validated and reliable test for young people. Body composition assessment was done by a tetrapolar bioimpedance in a fasting state and in the folicular phase for women. A maximal cardiopulmonary exercise testing, as well as the ST, evaluated the oxygen uptake at the peak of the test (VO2peak) by an ergospirometer Oxycon Mobile. Lipids, glucose, insulin were analysed and the ELISA method quantified the serum leptin and adiponectin from blood samples. Volunteers were divided in two groups: AR or MH according to a HI cutoff of 1.95, which was previously determined in the literature. T-test for comparison between groups, Pearson´s test to correlate main variables and ROC analysis for discriminating AR from up-and-down cycles in ST (SC) were applied (p<0.05). Results: Higher LA, fat mass (FM) and lower HDL, SC, leg lean mass (LM) and VO2peak were found in AR than in MH. Significant correlations were found between VO2peak and SC (r= 0.80) as well as between LA and FM (r=0.87), VO2peak (r=-0.73), and SC (r=-0.65). Area under de curve showed moderate accuracy (0.75) of SC <173 to discriminate AR phenotype. Conclusion: Our study found that at risk obese and normal-weight subjects showed an unhealthy metabolism as well as a poor CRF and functional daily activity capacity. Additionally, a simple and less costly functional test associated with above-mentioned aspects is able to identify ‘at risk’ subjects for primary intervention with important clinical and health implications.

Keywords: aerobic capacity, exercise, fitness, metabolism, obesity, 6MST

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53 Increased Envy and Schadenfreude in Parents of Newborns

Authors: Ana-María Gómez-Carvajal, Hernando Santamaría-García, Mateo Bernal, Mario Valderrama, Daniela Lizarazo, Juliana Restrepo, María Fernanda Barreto, Angélica Parra, Paula Torres, Diana Matallana, Jaime Silva, José Santamaría-García, Sandra Baez

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Higher levels of oxytocin are associated with better performance on social cognition tasks. However, higher levels of oxytocin have also been associated with increased levels of envy and schadenfreude. Considering these antecedents, this study aims to explore social emotions (i.e., envy and schadenfreude) and other components of social cognition (i.e. ToM and empathy), in women in the puerperal period and their respective partners, compared to a control group of men and women without children or partners. Control women should be in the luteal phase of the menstrual cycle or taking oral contraceptives as they allow oxytocin levels to remain stable. We selected this population since increased levels of oxytocin are present in both mothers and fathers of newborn babies. Both groups were matched by age, sex, and education level. Twenty-two parents of newborns (11 women, 11 men) and 15 controls (8 women, 7 men) performed an experimental task designed to trigger schadenfreude and envy. In this task, each participant was shown a real-life photograph and a description of two target characters matched in age and gender with the participant. The task comprised two experimental blocks. In the first block, participants read 15 sentences describing fortunate events involving either character. After reading each sentence, participants rated the event in terms of how much envy they felt for the character (1=no envy, 9=extreme envy). In the second block, participants read and reported the intensity of their pleasure (schadenfreude, 1=no pleasure, 9=extreme pleasure) in response to 15 unfortunate events happening to the characters. Five neutral events were included in each block. Moreover, participants were assessed with ToM and empathy tests. Potential confounding variables such as general cognitive functioning, stress levels, hours of sleep and depression symptoms were also measured. Results showed that parents of newborns showed increased levels of envy and schadenfreude. These effects are not explained by any confounding factor. Moreover, no significant differences were found in ToM or empathy tests. Our results offer unprecedented evidence of specific differences in envy and schadenfreude levels in parents of newborns. Our findings support previous studies showing a negative relationship between oxytocin levels and negative social emotions. Further studies should assess the direct relationship between oxytocin levels in parents of newborns and the performance in social emotions tasks.

Keywords: envy, empathy, oxytocin, schadenfreude, social emotions, theory of mind

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52 Comparison of Two Methods of Cryopreservation of Testicular Tissue from Prepubertal Lambs

Authors: Rensson Homero Celiz Ygnacio, Marco Aurélio Schiavo Novaes, Lucy Vanessa Sulca Ñaupas, Ana Paula Ribeiro Rodrigues

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The cryopreservation of testicular tissue emerges as an alternative for the preservation of the reproductive potential of individuals who still cannot produce sperm; however, they will undergo treatments that may affect their fertility (e.g., chemotherapy). Therefore, the present work aims to compare two cryopreservation methods (slow freezing and vitrification) in testicular tissue of prepubertal lambs. For that, to obtain the testicular tissue, the animals were castrated and the testicles were collected immediately in a physiological solution supplemented with antibiotics. In the laboratory, the testis was split into small pieces. The total size of the testicular fragments was 3×3x1 mm³ and was placed in a dish contained in Minimum Essential Medium (MEM-HEPES). The fragments were distributed randomly into non-cryopreserved (fresh control), slow freezing (SF), and vitrified. To SF procedures, two fragments from a given male were then placed in a 2,0 mL cryogenic vial containing 1,0 mL MEM-HEPES supplemented with 20% fetal bovine serum (FBS) and 20% dimethylsulfoxide (DMSO). Tubes were placed into a Mr. Frosty™ Freezing container with isopropyl alcohol and transferred to a -80°C freezer for overnight storage. On the next day, each tube was plunged into liquid nitrogen (NL). For vitrification, the ovarian tissue cryosystem (OTC) device was used. Testicular fragments were placed in the OTC device and exposed to the first vitrification solution composed of MEM-HEPES supplemented with 10 mg/mL Bovine Serum Albumin (BSA), 0.25 M sucrose, 10% Ethylene glycol (EG), 10% DMSO and 150 μM alpha-lipoic acid for four min. The VS1 was discarded and then the fragments were submerged into a second vitrification solution (VS2) containing the same composition of VS1 but 20% EG and 20% DMSO. VS2 was then discarded and each OTC device containing up to four testicular fragments was closed and immersed in NL. After the storage period, the fragments were removed from the NL, kept at room temperature for one min and then immersed at 37 °C in a water bath for 30 s. Samples were warmed by sequentially immersing in solutions of MEM-HEPES supplemented with 3 mg/mL BSA and decreasing concentrations of sucrose. Hematoxylin-eosin staining to analyze the tissue architecture was used. The score scale used was from 0 to 3, classified with a score 0 representing normal morphologically, and 3 were considered a lot of alteration. The histomorphological evaluation of the testicular tissue shows that when evaluating the nuclear alteration (distinction of nucleoli and condensation of nuclei), there are no differences when using slow freezing with respect to the control. However, vitrification presents greater damage (p <0.05). On the other hand, when evaluating the epithelial alteration, we observed that the freezing showed scores statistically equal to the control in variables such as retraction of the basement membrane, formation of gaps and organization of the peritubular cells. The results of the study demonstrated that cryopreservation using the slow freezing method is an excellent tool for the preservation of pubertal testicular tissue.

Keywords: cryopreservation, slow freezing, vitrification, testicular tissue, lambs

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51 Advanced Palliative Aquatics Care Multi-Device AuBento for Symptom and Pain Management by Sensorial Integration and Electromagnetic Fields: A Preliminary Design Study

Authors: J. F. Pollo Gaspary, F. Peron Gaspary, E. M. Simão, R. Concatto Beltrame, G. Orengo de Oliveira, M. S. Ristow Ferreira, J.C. Mairesse Siluk, I. F. Minello, F. dos Santos de Oliveira

Abstract:

Background: Although palliative care policies and services have been developed, research in this area continues to lag. An integrated model of palliative care is suggested, which includes complementary and alternative services aimed at improving the well-being of patients and their families. The palliative aquatics care multi-device (AuBento) uses several electromagnetic techniques to decrease pain and promote well-being through relaxation and interaction among patients, specialists, and family members. Aim: The scope of this paper is to present a preliminary design study of a device capable of exploring the various existing theories on the biomedical application of magnetic fields. This will be achieved by standardizing clinical data collection with sensory integration, and adding new therapeutic options to develop an advanced palliative aquatics care, innovating in symptom and pain management. Methods: The research methodology was based on the Work Package Methodology for the development of projects, separating the activities into seven different Work Packages. The theoretical basis was carried out through an integrative literature review according to the specific objectives of each Work Package and provided a broad analysis, which, together with the multiplicity of proposals and the interdisciplinarity of the research team involved, generated consistent and understandable complex concepts in the biomedical application of magnetic fields for palliative care. Results: Aubento ambience was idealized with restricted electromagnetic exposure (avoiding data collection bias) and sensory integration (allowing relaxation associated with hydrotherapy, music therapy, and chromotherapy or like floating tank). This device has a multipurpose configuration enabling classic or exploratory options on the use of the biomedical application of magnetic fields at the researcher's discretion. Conclusions: Several patients in diverse therapeutic contexts may benefit from the use of magnetic fields or fluids, thus validating the stimuli to clinical research in this area. A device in controlled and multipurpose environments may contribute to standardizing research and exploring new theories. Future research may demonstrate the possible benefits of the aquatics care multi-device AuBento to improve the well-being and symptom control in palliative care patients and their families.

Keywords: advanced palliative aquatics care, magnetic field therapy, medical device, research design

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50 A Review on Agricultural Landscapes as a Habitat of Rodents

Authors: Nadeem Munawar, Tariq Mahmood, Paula Rivadeneira, Ali Akhter

Abstract:

In this paper, we review on rodent species which are common inhabitants of agricultural landscapes where they are an important prey source for a wide variety of avian, reptilian, and mammalian predators. Agricultural fields are surrounded by fallow land, which provide suitable sites for shelter and breeding for rodents, while shrubs, grasses, annual weeds and forbs may provide supplementary food. The assemblage of rodent’s fauna in the cropland habitats including cropped fields, meadows and adjacent field structures like hedgerows, woodland and field margins fluctuates seasonally. The mature agricultural crops provides good source of food and shelter to the rodents and these factors along with favorable climatic factors/season facilitate breeding activities of these rodent species. Changes in vegetation height and vegetative cover affect two important aspects of a rodent’s life: food and shelter. In addition, during non-crop period vegetation can be important for building nests above or below ground and it provides thermal protection for rodents from heat and cold. The review revealed that rodents form a very diverse group of mammals, ranging from tiny pigmy mice to big capybaras, from arboreal flying squirrels to subterranean mole rats, from opportunistic omnivores (e.g. Norway rats) to specialist feeders (e.g. the North African fat sand rats that feed on a single family of plants only). It is therefore no surprise that some species thrive well under the conditions that are found in agricultural fields. The review on the population dynamics of the rodent species indicated that they are agricultural pests probably due to the heterogeneous landscape and to the high rotativity of vegetable crop cultivation. They also cause damage to various crops, directly and indirectly, by gnawing, spoilage, contamination and hoarding activities, besides this behavior they have also significance importance in agricultural habitat. The burrowing activities of rodents alter the soil properties around their burrows which improve its aeration, infiltration, increase the water holding capacity and thus encourage plant growth. These properties are beneficial for the soil because they affect absorption of phosphorus, absorption zinc, copper, other nutrients and the uptake of water and thus rodents are known as indicator species in agricultural fields. Our review suggests that wide crop field’s borders, particularly those contiguous to various cropland fields, should be understood as priority sites for nesting, feeding, and cover for the rodent’s fauna. The goal of this review paper is to provide a comprehensive synthesis of understanding regarding rodent habitat and biodiversity in agricultural landscapes.

Keywords: agricultural landscapes, food, indicator species, shelter

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49 Oncolytic H-1 Parvovirus Entry in Cancer Cells through Clathrin-Mediated Endocytosis

Authors: T. Ferreira, A. Kulkarni, C. Bretscher, K. Richter, M. Ehrlich, A. Marchini

Abstract:

H-1 protoparvovirus (H-1PV) is a virus with inherent oncolytic and oncosuppressive activities while remaining non-pathogenic in humans. H-1PV was the first oncolytic parvovirus to undergo clinical testing. Results from trials in patients with glioblastoma or pancreatic carcinoma showed an excellent safety profile and first signs of efficacy. H-1PV infection is vastly dependent on cellular factors, from cell attachment and entry to viral replication and egress. Hence, we believe that the characterisation of the parvovirus life cycle would ultimately help further improve H-1PV clinical outcome. In the present study, we explored the entry pathway of H-1PV in cervical HeLa and glioma NCH125 cancer cell lines. Electron and confocal microscopy showed viral particles associated with clathrin-coated pits and vesicles, providing the first evidence that H-1PV cell entry occurs through clathrin-mediated endocytosis. Accordingly, we observed that by blocking clathrin-mediated endocytosis with hypertonic sucrose, chlorpromazine, or pitstop 2, H-1PV transduction was markedly decreased. Accordingly, siRNA-mediated knockdown of AP2M1, which retains a crucial role in clathrin-mediated endocytosis, verified the reliance of H-1PV on this route to enter HeLa and NCH125 cancer cells. By contrast, we found no evidence of viral entry through caveolae-mediated endocytosis. Indeed, pre-treatment of cells with nystatin or methyl-β-cyclodextrin, both inhibitors of caveolae-mediated endocytosis, did not affect viral transduction levels. Unexpectedly, siRNA-mediated knockdown of caveolin-1, the main driver of caveolae-mediated endocytosis, increased H-1PV transduction, suggesting caveolin-1 is a negative modulator of H-1PV infection. We also show that H-1PV entry is dependent on dynamin, a protein responsible for mediating the scission of vesicle neck and promoting further internalisation. Furthermore, since dynamin inhibition almost completely abolished H-1PV infection, makes it unlikely that H-1PV uses macropinocytosis as an alternative pathway to enter cells. After viral internalisation, H-1PV passes through early to late endosomes as observed by confocal microscopy. Inside these endocytic compartments, the acidic environment proved to be crucial for a productive infection. Inhibition of acidification of pH dramatically reduced H-1PV transduction. Besides, a fraction of H-1PV particles was observed inside LAMP1-positive lysosomes, most likely following a non-infectious route. To the author's best knowledge, this is the first study to characterise the cell entry pathways of H-1PV. Along these lines, this work will further contribute to understand H-1PV oncolytic properties as well as to improve its clinical potential in cancer virotherapy.

Keywords: clathrin-mediated endocytosis, H-1 parvovirus, oncolytic virus, virus entry

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48 Cloning and Expression a Gene of β-Glucosidase from Penicillium echinulatum in Pichia pastoris

Authors: Amanda Gregorim Fernandes, Lorena Cardoso Cintra, Rosalia Santos Amorim Jesuino, Fabricia Paula De Faria, Marcio José Poças Fonseca

Abstract:

Bioethanol is one of the most promising biofuels and able to replace fossil fuels and reduce its different environmental impacts and can be generated from various agroindustrial waste. The Brazil is in first place in bioethanol production to be the largest producer of sugarcane. The bagasse sugarcane (SCB) has lignocellulose which is composed of three major components: cellulose, hemicellulose and lignin. Cellulose is a homopolymer of glucose units connected by glycosidic linkages. Among all species of Penicillium, Penicillium echinulatum has been the focus of attention because they produce high quantities of cellulase and the mutant strain 9A02S1 produces higher enzyme levels compared to the wild. Among the cellulases, the cellobiohydrolases enzymes are the main components of the cellulolytic system of fungi, and are also responsible for most of the potential hydrolytic in enzyme cocktails for the industrial processing of plant biomass and several cellobiohydrolases Penicillium had higher specific activity against cellulose compared to CBH I from Trichoderma reesei. This fact makes it an interesting pattern for higher yields in the enzymatic hydrolysis, and also they are important enzymes in the hydrolysis of crystalline regions of cellulose. Therefore, finding new and more active enzymes become necessary. Meanwhile, β-glycosidases act on soluble substrates and are highly dependent on cellobiohydrolases and endoglucanases action to provide the substrate in the hydrolysis of the biomass, but the cellobiohydrolases and endoglucanases are highly dependent β-glucosidases to maintain efficient hydrolysis. Thus, there is a need to understand the structure-function relationships that govern the catalytic activity of cellulolytic enzymes to elucidate its mechanism of action and optimize its potential as industrial biocatalysts. To evaluate the enzyme β-glucosidase of Penicillium echinulatum (PeBGL1) the gene was synthesized from the assembly sequence from a library in induction conditions and then the PeBGL1 gene was cloned in the vector pPICZαA and transformed into P. pastoris GS115. After processing, the producers of PeBGL1 were analyzed for enzyme activity and protein profile where a band of approximately 100 kDa was viewed. It was also carried out the zymogram. In partial characterization it was determined optimum temperature of 50°C and optimum pH of 6,5. In addition, to increase the secreted recombinant PeBGL1 production by Pichia pastoris, three parameters of P. pastoris culture medium were analysed: methanol, nitrogen source concentrations and the inoculum size. A 23 factorial design was effective in achieving the optimum condition. Altogether, these results point to the potential application of this P. echinulatum β-glucosidase in hydrolysis of cellulose for the production of bioethanol.

Keywords: bioethanol, biotechnology, beta-glucosidase, penicillium echinulatum

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47 Neonatal Sepsis in Dogs Attend in Veterinary Hospital of the Sao Paulo State University, Botucatu, Brazil – Incidence, Clinical Aspects and Mortality

Authors: Maria Lucia G. Lourenco, Keylla H. N. P. Pereira, Vivane Y. Hibaru, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

Abstract:

Neonatal sepsis is a systemic response to the acute generalized infection caused by one or more bacterial agents, representing the main infectious cause of neonatal mortality in dogs during the first three weeks of life. This study aims to describe the incidence of sepsis in neonate dogs, as well as the main clinical signs and mortality rates. The study included 735 neonates admitted to the Sao Paulo State University (UNESP) Veterinary Hospital, Botucatu, Sao Paulo, Brazil, between January 2018 and November 2019. Seven hundred thirty-five neonates, 14% (98/703) presented neonatal sepsis. The main sources of infection for the neonates were intrauterine (72.5%, 71/98), lactogenic (13.2%, 13/98), umbilical (5.1%, 5/98) and unidentified sources (9.2%, 9/98). The main non-specific clinical signs observed in the newborns were weakness, depression, impaired or absent reflexes, hypothermia, hypoglycemia, dehydration, reduced muscle tonus and diarrhea. The newborns also manifested clinical signs of severe infection, such as hyperemia in the abdominal and anal regions, omphalitis, hematuria, abdomen and extremities with purplish-blue coloration necrosing injuries in the pads, bradycardia, dyspnea, epistaxis, hypotension and evolution to septic shock. Infections acquired during intrauterine life led to the onset of the clinical signs at the time of birth, with fast evolution during the first hours of life. On the other hand, infections acquired via milk or umbilical cord presented clinical signs later. The total mortality rate was 5.4% (38/703) and the mortality rate among the neonates with sepsis was 38.7% (38/98). The early mortality rate (0 to 2 days) accounted for 86.9% (33/38) and the late mortality rate (3 to 30 days) for 13.1% (5/38) of the deaths among the newborns with sepsis. The main bacterial agents observed were Staphylococcus spp., Streptococcus spp., Proteus spp. Mannheimia spp. and Escherichia coli. Neonatal sepsis evolves quickly and may lead to high mortality in a litter. The prognosis is usually favorable if the diagnosis is reached early and the antibiotic therapy instituted as soon as possible, even before the results of blood cultures and antibiograms. The therapeutic recommendations should meet the special physiological conditions of a neonate in terms of metabolism and excretion of medication. Therefore, it is of utmost importance that the veterinarian is knowledgeable regarding neonatology to provide effective intervention and improve the survival rates of these patients.

Keywords: Neonatal infection , bacteria, puppies, newborn

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46 Foreseen the Future: Human Factors Integration in European Horizon Projects

Authors: José Manuel Palma, Paula Pereira, Margarida Tomás

Abstract:

Foreseen the future: Human factors integration in European Horizon Projects The development of new technology as artificial intelligence, smart sensing, robotics, cobotics or intelligent machinery must integrate human factors to address the need to optimize systems and processes, thereby contributing to the creation of a safe and accident-free work environment. Human Factors Integration (HFI) consistently pose a challenge for organizations when applied to daily operations. AGILEHAND and FORTIS projects are grounded in the development of cutting-edge technology - industry 4.0 and 5.0. AGILEHAND aims to create advanced technologies for autonomously sort, handle, and package soft and deformable products, whereas FORTIS focuses on developing a comprehensive Human-Robot Interaction (HRI) solution. Both projects employ different approaches to explore HFI. AGILEHAND is mainly empirical, involving a comparison between the current and future work conditions reality, coupled with an understanding of best practices and the enhancement of safety aspects, primarily through management. FORTIS applies HFI throughout the project, developing a human-centric approach that includes understanding human behavior, perceiving activities, and facilitating contextual human-robot information exchange. it intervention is holistic, merging technology with the physical and social contexts, based on a total safety culture model. In AGILEHAND we will identify safety emergent risks, challenges, their causes and how to overcome them by resorting to interviews, questionnaires, literature review and case studies. Findings and results will be presented in “Strategies for Workers’ Skills Development, Health and Safety, Communication and Engagement” Handbook. The FORTIS project will implement continuous monitoring and guidance of activities, with a critical focus on early detection and elimination (or mitigation) of risks associated with the new technology, as well as guidance to adhere correctly with European Union safety and privacy regulations, ensuring HFI, thereby contributing to an optimized safe work environment. To achieve this, we will embed safety by design, and apply questionnaires, perform site visits, provide risk assessments, and closely track progress while suggesting and recommending best practices. The outcomes of these measures will be compiled in the project deliverable titled “Human Safety and Privacy Measures”. These projects received funding from European Union’s Horizon 2020/Horizon Europe research and innovation program under grant agreement No101092043 (AGILEHAND) and No 101135707 (FORTIS).

Keywords: human factors integration, automation, digitalization, human robot interaction, industry 4.0 and 5.0

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45 Functional Analysis of Variants Implicated in Hearing Loss in a Cohort from Argentina: From Molecular Diagnosis to Pre-Clinical Research

Authors: Paula I. Buonfiglio, Carlos David Bruque, Lucia Salatino, Vanesa Lotersztein, Sebastián Menazzi, Paola Plazas, Ana Belén Elgoyhen, Viviana Dalamón

Abstract:

Hearing loss (HL) is the most prevalent sensorineural disorder affecting about 10% of the global population, with more than half due to genetic causes. About 1 in 500-1000 newborns present congenital HL. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance. To date, more than 100 genes are related to HL. Therefore, the Whole-exome sequencing (WES) technique has become a cost-effective alternative approach for molecular diagnosis. Nevertheless, new challenges arise from the detection of novel variants, in particular missense changes, which can lead to a spectrum of genotype-to-phenotype correlations, which is not always straightforward. In this work, we aimed to identify the genetic causes of HL in isolated and familial cases by designing a multistep approach to analyze target genes related to hearing impairment. Moreover, we performed in silico and in vivo analyses in order to further study the effect of some of the novel variants identified in the hair cell function using the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes, respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Fifty of the undiagnosed patients with moderate HL were tested for deletions in STRC gene by Multiplex ligation-dependent probe amplification technique (MLPA), leading to 6% of diagnosis. After this initial screening, 50 families were selected to be analyzed by WES, achieving diagnosis in 44% of them. Half of the identified variants were novel. A missense variant in MYO6 gene detected in a family with postlingual HL was selected to be further analyzed. A protein modeling with AlphaFold2 software was performed, proving its pathogenic effect. In order to functionally validate this novel variant, a knockdown phenotype rescue assay in zebrafish was carried out. Injection of wild-type MYO6 mRNA in embryos rescued the phenotype, whereas using the mutant MYO6 mRNA (carrying c.2782C>A variant) had no effect. These results strongly suggest the deleterious effect of this variant on the mobility of stereocilia in zebrafish neuromasts, and hence on the auditory system. In the present work, we demonstrated that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. These results highlight the importance of a combined strategy in order to identify candidate variants as well as the in silico and in vivo studies to analyze and prove their pathogenicity and accomplish a better understanding of the mechanisms underlying the physiopathology of the hearing impairment.

Keywords: diagnosis, genetics, hearing loss, in silico analysis, in vivo analysis, WES, zebrafish

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44 Polymer Nanocomposite Containing Silver Nanoparticles for Wound Healing

Authors: Patrícia Severino, Luciana Nalone, Daniele Martins, Marco Chaud, Classius Ferreira, Cristiane Bani, Ricardo Albuquerque

Abstract:

Hydrogels produced with polymers have been used in the development of dressings for wound treatment and tissue revitalization. Our study on polymer nanocomposites containing silver nanoparticles shows antimicrobial activity and applications in wound healing. The effects are linked with the slow oxidation and Ag⁺ liberation to the biological environment. Furthermore, bacterial cell membrane penetration and metabolic disruption through cell cycle disarrangement also contribute to microbial cell death. The silver antimicrobial activity has been known for many years, and previous reports show that low silver concentrations are safe for human use. This work aims to develop a hydrogel using natural polymers (sodium alginate and gelatin) combined with silver nanoparticles for wound healing and with antimicrobial properties in cutaneous lesions. The hydrogel development utilized different sodium alginate and gelatin proportions (20:80, 50:50 and 80:20). The silver nanoparticles incorporation was evaluated at the concentrations of 1.0, 2.0 and 4.0 mM. The physico-chemical properties of the formulation were evaluated using ultraviolet-visible (UV-Vis) absorption spectroscopy, Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), and thermogravimetric (TG) analysis. The morphological characterization was made using transmission electron microscopy (TEM). Human fibroblast (L2929) viability assay was performed with a minimum inhibitory concentration (MIC) assessment as well as an in vivo cicatrizant test. The results suggested that sodium alginate and gelatin in the (80:20) proportion with 4 mM of AgNO₃ in the (UV-Vis) exhibited a better hydrogel formulation. The nanoparticle absorption spectra of this analysis showed a maximum band around 430 - 450 nm, which suggests a spheroidal form. The TG curve exhibited two weight loss events. DSC indicated one endothermic peak at 230-250 °C, due to sample fusion. The polymers acted as stabilizers of a nanoparticle, defining their size and shape. Human fibroblast viability assay L929 gave 105 % cell viability with a negative control, while gelatin presented 96% viability, alginate: gelatin (80:20) 96.66 %, and alginate 100.33 % viability. The sodium alginate:gelatin (80:20) exhibited significant antimicrobial activity, with minimal bacterial growth at a ratio of 1.06 mg.mL⁻¹ in Pseudomonas aeruginosa and 0.53 mg.mL⁻¹ in Staphylococcus aureus. The in vivo results showed a significant reduction in wound surface area. On the seventh day, the hydrogel-nanoparticle formulation reduced the total area of injury by 81.14 %, while control reached a 45.66 % reduction. The results suggest that silver-hydrogel nanoformulation exhibits potential for wound dressing therapeutics.

Keywords: nanocomposite, wound healing, hydrogel, silver nanoparticle

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43 Assessment of Commercial Antimicrobials Incorporated into Gelatin Coatings and Applied to Conventional Heat-Shrinking Material for the Prevention of Blown Pack Spoilage in Vacuum Packaged Beef Cuts

Authors: Andrey A. Tyuftin, Rachael Reid, Paula Bourke, Patrick J. Cullen, Seamus Fanning, Paul Whyte, Declan Bolton , Joe P. Kerry

Abstract:

One of the primary spoilage issues associated with vacuum-packed beef products is blown pack spoilage (BPS) caused by the psychrophilic spore-forming strain of Clostridium spp. Spores derived from this organism can be activated after heat-shrinking (eg. 90°C for 3 seconds). To date, research into the control of Clostridium spp in beef packaging is limited. Active packaging in the form of antimicrobially-active coatings may be one approach to its control. Antimicrobial compounds may be incorporated into packaging films or coated onto the internal surfaces of packaging films using a carrier matrix. Three naturally-sourced, commercially-available antimicrobials, namely; Auranta FV (AFV) (bitter oranges extract) from Envirotech Innovative Products Ltd, Ireland; Inbac-MDA (IMDA) from Chemital LLC, Spain, mixture of different organic acids and sodium octanoate (SO) from Sigma-Aldrich, UK, were added into gelatin solutions at 2 concentrations: 2.5 and 3.5 times their minimum inhibition concentration (MIC) against Clostridium estertheticum (DSMZ 8809). These gelatin solutions were coated onto the internal polyethylene layer of cold plasma treated, heat-shrinkable laminates conventionally used for meat packaging applications. Atmospheric plasma was used in order to enhance adhesion between packaging films and gelatin coatings. Pouches were formed from these coated packaging materials, and beef cuts which had been inoculated with C. estertheticum were vacuum packaged. Inoculated beef was vacuum packaged without employing active films and this treatment served as the control. All pouches were heat-sealed and then heat-shrunk at 90°C for 3 seconds and incubated at 2°C for 100 days. During this storage period, packs were monitored for the indicators of blown pack spoilage as follows; gas bubbles in drip, loss of vacuum (onset of BPS), blown, the presence of sufficient gas inside the packs to produce pack distension and tightly stretched, “overblown” packs/ packs leaking. Following storage and assessment of indicator date, it was concluded that AFV- and SO-containing packaging inhibited the growth of C. estertheticum, significantly delaying the blown pack spoilage of beef primals. IMDA did not inhibit the growth of C. estertheticum. This may be attributed to differences in release rates and possible reactions with gelatin. Overall, active films were successfully produced following plasma surface treatment, and experimental data demonstrated clearly that the use of antimicrobially-active films could significantly prolong the storage stability of beef primals through the effective control of BPS.

Keywords: active packaging, blown pack spoilage, Clostridium, antimicrobials, edible coatings, food packaging, gelatin films, meat science

Procedia PDF Downloads 236
42 Quality by Design in the Optimization of a Fast HPLC Method for Quantification of Hydroxychloroquine Sulfate

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

Abstract:

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

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

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41 Procedure for Monitoring the Process of Behavior of Thermal Cracking in Concrete Gravity Dams: A Case Study

Authors: Adriana de Paula Lacerda Santos, Bruna Godke, Mauro Lacerda Santos Filho

Abstract:

Several dams in the world have already collapsed, causing environmental, social and economic damage. The concern to avoid future disasters has stimulated the creation of a great number of laws and rules in many countries. In Brazil, Law 12.334/2010 was created, which establishes the National Policy on Dam Safety. Overall, this policy requires the dam owners to invest in the maintenance of their structures and to improve its monitoring systems in order to provide faster and straightforward responses in the case of an increase of risks. As monitoring tools, visual inspections has provides comprehensive assessment of the structures performance, while auscultation’s instrumentation has added specific information on operational or behavioral changes, providing an alarm when a performance indicator exceeds the acceptable limits. These limits can be set using statistical methods based on the relationship between instruments measures and other variables, such as reservoir level, time of the year or others instruments measuring. Besides the design parameters (uplift of the foundation, displacements, etc.) the dam instrumentation can also be used to monitor the behavior of defects and damage manifestations. Specifically in concrete gravity dams, one of the main causes for the appearance of cracks, are the concrete volumetric changes generated by the thermal origin phenomena, which are associated with the construction process of these structures. Based on this, the goal of this research is to propose a monitoring process of the thermal cracking behavior in concrete gravity dams, through the instrumentation data analysis and the establishment of control values. Therefore, as a case study was selected the Block B-11 of José Richa Governor Dam Power Plant, that presents a cracking process, which was identified even before filling the reservoir in August’ 1998, and where crack meters and surface thermometers were installed for its monitoring. Although these instruments were installed in May 2004, the research was restricted to study the last 4.5 years (June 2010 to November 2014), when all the instruments were calibrated and producing reliable data. The adopted method is based on simple linear correlations procedures to understand the interactions among the instruments time series, verifying the response times between them. The scatter plots were drafted from the best correlations, which supported the definition of the limit control values. Among the conclusions, it is shown that there is a strong or very strong correlation between ambient temperature and the crack meters and flowmeters measurements. Based on the results of the statistical analysis, it was possible to develop a tool for monitoring the behavior of the case study cracks. Thus it was fulfilled the goal of the research to develop a proposal for a monitoring process of the behavior of thermal cracking in concrete gravity dams.

Keywords: concrete gravity dam, dams safety, instrumentation, simple linear correlation

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40 Anabasine Intoxication and Its Relation to Plant Develoment Stages

Authors: Thaís T. Valério Caetano, Lívia de Carvalho Ferreira, João Máximo De Siqueira, Carlos Alexandre Carollo, Arthur Ladeira Macedo, Vanessa C. Stein

Abstract:

Nicotiana glauca, commonly known as wild tobacco or tobacco bush, belongs to the Solanaceae family. It is native to South America but has become naturalized in various regions, including Australia, California, Africa, and the Mediterranean. N. glauca is listed in the Global Invasive Species Database (GISD) and the Invasive Species Compendium (CABI). It is known for producing pyridine alkaloids, including anabasine, which is highly toxic. Anabasine is predominantly found in the leaves and can cause severe health issues such as neuromuscular blockade, respiratory arrest, and cardiovascular problems when ingested. Mistaken identity with edible plants like spinach has resulted in food poisoning cases in Israel and Brazil. Anabasine, a minor alkaloid constituent of tobacco, may contribute to tobacco addiction by mimicking or enhancing the effects of nicotine. Therefore, it is essential to investigate the production pattern of anabasine and its relationship to the developmental stages of the plant. This study aimed to establish the relationship between the phenological plant age, cultivation place, and the increase in anabasine concentration, which can lead to human intoxication cases. In this study, N. glauca plants were collected from three different rural areas in Brazil during a year to examine leaves at various stages of development. Samples were also obtained from cultivated plants in Marilândia, Minas Gerais, Brazil, as well as from Divinópolis, Minas Gerais, Brazil, and Arraial do Cabo, Rio de Janeiro, Brazil. In vitro cultivated plants on MS medium were included in the study. The collected leaves were dried, powdered, and stored. Alkaloid extraction was performed using a methanol and water mixture, followed by liquid-liquid extraction with chloroform. The anabasine content was determined using HPLC-DAD analysis with nicotine as a standard. The results indicated that anabasine production increases with the plant's development, peaking in adult leaves during the reproduction phase and declining afterward. In vitro, plants showed similar anabasine production to young leaves. The successful adaptation of N. glauca in new environments poses a global problem, and the correlation between anabasine production and the plant's developmental stages has been understudied. The presence of substances produced by the plant can pose a risk to other species, especially when mistaken for edible plants. The findings from this study shed light on the pattern of anabasine production and its association with plant development, contributing to a better understanding of the potential risks associated with N. glauca and the importance of accurate identification.

Keywords: alkaloid production, invasive species, nicotiana glauca, plant phenology

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39 Application of NBR 14861: 2011 for the Design of Prestress Hollow Core Slabs Subjected to Shear

Authors: Alessandra Aparecida Vieira França, Adriana de Paula Lacerda Santos, Mauro Lacerda Santos Filho

Abstract:

The purpose of this research i to study the behavior of precast prestressed hollow core slabs subjected to shear. In order to achieve this goal, shear tests were performed using hollow core slabs 26,5cm thick, with and without a concrete cover of 5 cm, without cores filled, with two cores filled and three cores filled with concrete. The tests were performed according to the procedures recommended by FIP (1992), the EN 1168:2005 and following the method presented in Costa (2009). The ultimate shear strength obtained within the tests was compared with the values of theoretical resistant shear calculated in accordance with the codes, which are being used in Brazil, noted: NBR 6118:2003 and NBR 14861:2011. When calculating the shear resistance through the equations presented in NBR 14861:2011, it was found that provision is much more accurate for the calculation of the shear strength of hollow core slabs than the NBR 6118 code. Due to the large difference between the calculated results, even for slabs without cores filled, the authors consulted the committee that drafted the NBR 14861:2011 and found that there is an error in the text of the standard, because the coefficient that is suggested, actually presents the double value than the needed one! The ABNT, later on, soon issued an amendment of NBR 14861:2011 with the necessary corrections. During the tests for the present study, it was confirmed that the concrete filling the cores contributes to increase the shear strength of hollow core slabs. But in case of slabs 26,5 cm thick, the quantity should be limited to a maximum of two cores filled, because most of the results for slabs with three cores filled were smaller. This confirmed the recommendation of NBR 14861:2011which is consistent with standard practice. After analyzing the configuration of cracking and failure mechanisms of hollow core slabs during the shear tests, strut and tie models were developed representing the forces acting on the slab at the moment of rupture. Through these models the authors were able to calculate the tensile stress acting on the concrete ties (ribs) and scaled the geometry of these ties. The conclusions of the research performed are the experiments results have shown that the mechanism of failure of the hollow-core slabs can be predicted using the strut-and-tie procedure, within a good range of accuracy. In addition, the needed of the correction of the Brazilian standard to review the correction factor σcp duplicated (in NBR14861/2011), and the limitation of the number of cores (Holes) to be filled with concrete, to increase the strength of the slab for the shear resistance. It is also suggested the increasing the amount of test results with 26.5 cm thick, and a larger range of thickness slabs, in order to obtain results of shear tests with cores concreted after the release of prestressing force. Another set of shear tests on slabs must be performed in slabs with cores filled and cover concrete reinforced with welded steel mesh for comparison with results of theoretical values calculated by the new revision of the standard NBR 14861:2011.

Keywords: prestressed hollow core slabs, shear, strut, tie models

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38 Electrophysiological Correlates of Statistical Learning in Children with and without Developmental Language Disorder

Authors: Ana Paula Soares, Alexandrina Lages, Helena Oliveira, Francisco-Javier Gutiérrez-Domínguez, Marisa Lousada

Abstract:

From an early age, exposure to a spoken language allows us to implicitly capture the structure underlying the succession of the speech sounds in that language and to segment it into meaningful units (words). Statistical learning (SL), i.e., the ability to pick up patterns in the sensory environment even without intention or consciousness of doing it, is thus assumed to play a central role in the acquisition of the rule-governed aspects of language and possibly to lie behind the language difficulties exhibited by children with development language disorder (DLD). The research conducted so far has, however, led to inconsistent results, which might stem from the behavioral tasks used to test SL. In a classic SL experiment, participants are first exposed to a continuous stream (e.g., syllables) in which, unbeknownst to the participants, stimuli are grouped into triplets that always appear together in the stream (e.g., ‘tokibu’, ‘tipolu’), with no pauses between each other (e.g., ‘tokibutipolugopilatokibu’) and without any information regarding the task or the stimuli. Following exposure, SL is assessed by asking participants to discriminate between triplets previously presented (‘tokibu’) from new sequences never presented together during exposure (‘kipopi’), i.e., to perform a two-alternative-forced-choice (2-AFC) task. Despite the widespread use of the 2-AFC to test SL, it has come under increasing criticism as it is an offline post-learning task that only assesses the result of the learning that had occurred during the previous exposure phase and that might be affected by other factors beyond the computation of regularities embedded in the input, typically the likelihood two syllables occurring together, a statistic known as transitional probability (TP). One solution to overcome these limitations is to assess SL as exposure to the stream unfolds using online techniques such as event-related potentials (ERP) that is highly sensitive to the time-course of the learning in the brain. Here we collected ERPs to examine the neurofunctional correlates of SL in preschool children with DLD, and chronological-age typical language development (TLD) controls who were exposed to an auditory stream in which eight three-syllable nonsense words, four of which presenting high-TPs and the other four low-TPs, to further analyze whether the ability of DLD and TLD children to extract-word-like units from the steam was modulated by words’ predictability. Moreover, to ascertain if the previous knowledge of the to-be-learned-regularities affected the neural responses to high- and low-TP words, children performed the auditory SL task, firstly, under implicit, and, subsequently, under explicit conditions. Although behavioral evidence of SL was not obtained in either group, the neural responses elicited during the exposure phases of the SL tasks differentiated children with DLD from children with TLD. Specifically, the results indicated that only children from the TDL group showed neural evidence of SL, particularly in the SL task performed under explicit conditions, firstly, for the low-TP, and, subsequently, for the high-TP ‘words’. Taken together, these findings support the view that children with DLD showed deficits in the extraction of the regularities embedded in the auditory input which might underlie the language difficulties.

Keywords: development language disorder, statistical learning, transitional probabilities, word segmentation

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37 Association between Obstetric Factors with Affected Areas of Health-Related Quality of Life of Pregnant Women

Authors: Cinthia G. P. Calou, Franz J. Antezana, Ana I. O. Nicolau, Eveliny S. Martins, Paula R. A. L. Soares, Glauberto S. Quirino, Dayanne R. Oliveira, Priscila S. Aquino, Régia C. M. B. Castro, Ana K. B. Pinheiro

Abstract:

Introduction: As an integral part of the health-disease process, gestation is a period in which the social insertion of women can influence, in a positive or negative way, the course of the pregnancy-puerperal cycle. Thus, evaluating the quality of life of this population can redirect the implementation of innovative practices in the quest to make them more effective and real for the promotion of a more humanized care. This study explores the associations between the obstetric factors with affected areas of health-related quality of life of pregnant women with habitual risk. Methods: This is a cross-sectional, quantitative study conducted in three public facilities and a private service that provides prenatal care in the city of Fortaleza, Ceara, Brazil. The sample consisted of 261 pregnant women who underwent low-risk prenatal care and were interviewed from September to November 2014. The collection instruments were a questionnaire containing socio-demographic and obstetric variables, in addition to the Brazilian version of the Mother scale Generated Index (MGI) characterized by being a specific and objective instrument, consisting of a single sheet and subdivided into three stages. It allows identifying the areas of life of the pregnant woman that are most affected, which could go unnoticed by the pre-formulated measurement instruments. The obstetric data, as well as the data concerning the application of the MGI scale, were compiled and analyzed through the statistical program Statistical Package for the Social Sciences (SPSS), version 20.0. After the compilation, a descriptive analysis was carried out. Then, associations were made between some variables. The tests applied were the Pearson Chi-Square and the Fisher's exact test. The odds ratio was also calculated. These associations were considered statistically significant when the p (probability) value was less than or equal to a level of 5% (α = 0.05) in the tests performed. Results: The variables that negatively reflected the quality of life of the pregnant women and presented a significant association with the polaciuria were: gestational age (p = 0.022) and parity (p = 0.048). Episodes of nausea and vomiting also showed significant with gestational age correlation (p = 0.0001). Evaluating the crossing of stress, we observed a significant association with parity (p = 0.0001). In turn, emotional lability revealed dependence on the variable type of delivery (p = 0.009). Conclusion: The health professionals involved in the assistance to the pregnant woman can understand how the process of gestation is experienced, considering all its peculiar transformations; to meet their individual needs, stimulating their autonomy and their power of choice, envisaging the achievement of a better quality of life related to health in the perspective of health promotion.

Keywords: health-related quality of life, obstetric nursing, pregnant women, prenatal care

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36 Relationships of Plasma Lipids, Lipoproteins and Cardiovascular Outcomes with Climatic Variations: A Large 8-Year Period Brazilian Study

Authors: Vanessa H. S. Zago, Ana Maria H. de Avila, Paula P. Costa, Welington Corozolla, Liriam S. Teixeira, Eliana C. de Faria

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Objectives: The outcome of cardiovascular disease is affected by environment and climate. This study evaluated the possible relationships between climatic and environmental changes and the occurrence of biological rhythms in serum lipids and lipoproteins in a large population sample in the city of Campinas, State of Sao Paulo, Brazil. In addition, it determined the temporal variations of death due to atherosclerotic events in Campinas during the time window examined. Methods: A large 8-year retrospective study was carried out to evaluate the lipid profiles of individuals attended at the University of Campinas (Unicamp). The study population comprised 27.543 individuals of both sexes and of all ages. Normolipidemic and dyslipidemic individuals classified according to Brazilian guidelines on dyslipidemias, participated in the study. For the same period, the temperature, relative humidity and daily brightness records were obtained from the Centro de Pesquisas Meteorologicas e Climaticas Aplicadas a Agricultura/Unicamp and frequencies of death due to atherosclerotic events in Campinas were acquired from the Brazilian official database DATASUS, according to the International Classification of Diseases. Statistical analyses were performed using both Cosinor and ARIMA temporal analysis methods. For cross-correlation analysis between climatic and lipid parameters, cross-correlation functions were used. Results: Preliminary results indicated that rhythmicity was significant for LDL-C and HDL-C in the cases of both normolipidemic and dyslipidemic subjects (n =respectively 11.892 and 15.651 both measures increasing in the winter and decreasing in the summer). On the other hand, for dyslipidemic subjects triglycerides increased in summer and decreased in winter, in contrast to normolipidemic ones, in which triglycerides did not show rhythmicity. The number of deaths due to atherosclerotic events showed significant rhythmicity, with maximum and minimum frequencies in winter and summer, respectively. Cross-correlation analyzes showed that low humidity and temperature, higher thermal amplitude and dark cycles are associated with increased levels of LDL-C and HDL-C during winter. In contrast, TG showed moderate cross-correlations with temperature and minimum humidity in an inverse way: maximum temperature and humidity increased TG during the summer. Conclusions: This study showed a coincident rhythmicity between low temperatures and high concentrations of LDL-C and HDL-C and the number of deaths due to atherosclerotic cardiovascular events in individuals from the city of Campinas. The opposite behavior of cholesterol and TG suggest different physiological mechanisms in their metabolic modulation by climate parameters change. Thus, new analyses are underway to better elucidate these mechanisms, as well as variations in lipid concentrations in relation to climatic variations and their associations with atherosclerotic disease and death outcomes in Campinas.

Keywords: atherosclerosis, climatic variations, lipids and lipoproteins, associations

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35 Validation of an Educative Manual for Patients with Breast Cancer Submitted to Radiation Therapy

Authors: Flavia Oliveira de A. M. Cruz, Edison Tostes Faria, Paula Elaine D. Reis

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When the breast is submitted to radiation therapy (RT), the most common effects are pain, skin changes, mobility restrictions, local sensory alteration, and fatigue. These effects, if not managed properly, may reduce the quality of life of cancer patients and may lead to the treatment discontinuation. Therefore, promoting knowledge and guidelines for symptom management remain a high priority for patients and a challenge for health professionals, due to the need to handle side effects in a population with a life-threatening disease. Printed materials are important strategies for supporting educative activities since they help the individual to assimilate and understand the amount of information transmitted. Nurses' behavior can be systematized through the use of an educative manual, which may be effective in promoting information regarding the treatment, self-care and how to control the effects of RT at home. In view of the importance of guaranteeing the validity of the material before its use, the objective of this research was to validate the content and appearance of an educative manual for breast cancer patients undergoing RT. The Theory of Psychometrics was used for the validation process in this descriptive methodological research. A minimum agreement rate (AR) of 80% was considered to guarantee the validity of the material. The data were collected from October to December 2017, by means of two assessments tools, constructed in the form of a Likert scale, with five levels of understanding. These instruments addressed different aspects of the evaluation, in view of two different groups of participants; 17 experts in the theme area of the educative manual, and 12 women that received RT previously to treat breast cancer. The manual was titled 'Orientation Manual: radiation therapy in breast', and was focused on breast cancer patients attended at the Department of Oncology of the Brasília University Hospital (UNACON/HUB). The research project was submitted to the Research Ethics Committee at the School of Health Sciences of the University of Brasília (CAAE: 24592213.1.0000.0030). Only two items of the assessment tool for the experts, one related to the manual's ability to promote behavioral and attitude changes and the other related to the extent of its use for other health services, obtained AR < 80% and were reformulated based on the participants' suggestions and in the literature. All other items were considered appropriate and/or complete appropriate in the three blocks proposed for the experts: objectives - 89%, structure and form - 93%, and relevance - 93%; and good and/or very good in the five blocks of analysis proposed for patients: objectives - 100%, organization - 100%, writing style - 100%, appearance - 100%, and motivation. The appearance and content validation of the educative manual proposed were attended to. The educative manual was considered relevant and pertinent and may contribute to the understanding of the therapeutic process by breast cancer patients during RT, as well as support clinical practice through the nursing consultation.

Keywords: oncology nursing, nursing care, validation studies, educational technology

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34 Phenotypic and Molecular Heterogeneity Linked to the Magnesium Transporter CNNM2

Authors: Reham Khalaf-Nazzal, Imad Dweikat, Paula Gimenez, Iker Oyenarte, Alfonso Martinez-Cruz, Domonik Muller

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Metal cation transport mediator (CNNM) gene family comprises 4 isoforms that are expressed in various human tissues. Structurally, CNNMs are complex proteins that contain an extracellular N-terminal domain preceding a DUF21 transmembrane domain, a ‘Bateman module’ and a C-terminal cNMP-binding domain. Mutations in CNNM2 cause familial dominant hypomagnesaemia. Growing evidence highlights the role of CNNM2 in neurodevelopment. Mutations in CNNM2 have been implicated in epilepsy, intellectual disability, schizophrenia, and others. In the present study, we aim to elucidate the function of CNNM2 in the developing brain. Thus, we present the genetic origin of symptoms in two family cohorts. In the first family, three siblings of a consanguineous Palestinian family in which parents are first cousins, and consanguinity ran over several generations, presented a varying degree of intellectual disability, cone-rod dystrophy, and autism spectrum disorder. Exome sequencing and segregation analysis revealed the presence of homozygous pathogenic mutation in the CNNM2 gene, the parents were heterozygous for that gene mutation. Magnesium blood levels were normal in the three children and their parents in several measurements. They had no symptoms of hypomagnesemia. The CNNM2 mutation in this family was found to locate in the CBS1 domain of the CNNM2 protein. The crystal structure of the mutated CNNM2 protein was not significantly different from the wild-type protein, and the binding of AMP or MgATP was not dramatically affected. This suggests that the CBS1 domain could be involved in pure neurodevelopmental functions independent of its magnesium-handling role, and this mutation could have affected a protein partner binding or other functions in this protein. In the second family, another autosomal dominant CNNM2 mutation was found to run in a large family with multiple individuals over three generations. All affected family members had hypomagnesemia and hypermagnesuria. Oral supplementation of magnesium did not increase the levels of magnesium in serum significantly. Some affected members of this family have defects in fine motor skills such as dyslexia and dyslalia. The detected mutation is located in the N-terminal part, which contains a signal peptide thought to be involved in the sorting and routing of the protein. In this project, we describe heterogenous clinical phenotypes related to CNNM2 mutations and protein functions. In the first family, and up to the authors’ knowledge, we report for the first time the involvement of CNNM2 in retinal photoreceptor development and function. In addition, we report the presence of a neurophenotype independent of magnesium status related to the CNNM2 protein mutation. Taking into account the different modes of inheritance and the different positions of the mutations within CNNM2 and its different structural and functional domains, it is likely that CNNM2 might be involved in a wide spectrum of neuropsychiatric comorbidities with considerable varying phenotypes.

Keywords: magnesium transport, autosomal recessive, autism, neurodevelopment, CBS domain

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