Search results for: Denise Teixeira

Authors: Carolina N. França, Jônatas B. do Amaral, Maria C.O. Izar, Ighor L. Teixeira, Francisco A. Fonseca

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Introduction: Atherosclerosis is a progressive disease, characterized by lipid and fibrotic element deposition in large-caliber arteries. Conditions related to the development of atherosclerosis, as dyslipidemia, hypertension, diabetes, and smoking are associated with endothelial dysfunction. There is a frequent recurrence of cardiovascular outcomes after acute myocardial infarction and, at this sense, cycles of mobilization of monocyte subtypes (classical, intermediate and nonclassical) secondary to myocardial infarction may determine the colonization of atherosclerotic plaques in different stages of the development, contributing to early recurrence of ischemic events. The recruitment of different monocyte subsets during inflammatory process requires the expression of chemokine receptors CCR2, CCR5, and CX3CR1, to promote the migration of monocytes to the inflammatory site. The aim of this study was to evaluate the effect of lipid-lowering treatment by six months in the monocyte phenotype and chemokine receptor levels of patients after Acute Myocardial Infarction (AMI). Methods: This is a PROBE (prospective, randomized, open-label trial with blinded endpoints) study (ClinicalTrials.gov Identifier: NCT02428374). Adult patients (n=147) of both genders, ageing 18-75 years, were randomized in a 2x2 factorial design for treatment with rosuvastatin 20 mg/day or simvastatin 40 mg/day plus ezetimibe 10 mg/day as well as ticagrelor 90 mg 2x/day and clopidogrel 75 mg, in addition to conventional AMI therapy. Blood samples were collected at baseline, after one month and six months of treatment. Monocyte subtypes (classical - inflammatory, intermediate - phagocytic and nonclassical – anti-inflammatory) were identified, quantified and characterized by flow cytometry, as well as the expressions of the chemokine receptors (CCR2, CCR5 and CX3CR1) were also evaluated in the mononuclear cells. Results: After six months of treatment, there was an increase in the percentage of classical monocytes and reduction in the nonclassical monocytes (p=0.038 and p < 0.0001 Friedman Test), without differences for intermediate monocytes. Besides, classical monocytes had higher expressions of CCR5 and CX3CR1 after treatment, without differences related to CCR2 (p < 0.0001 for CCR5 and CX3CR1; p=0.175 for CCR2). Intermediate monocytes had higher expressions of CCR5 and CX3CR1 and lower expression of CCR2 (p = 0.003; p < 0.0001 and p = 0.011, respectively). Nonclassical monocytes had lower expressions of CCR2 and CCR5, without differences for CX3CR1 (p < 0.0001; p = 0.009 and p = 0.138, respectively). There were no differences after the comparison between the four treatment arms. Conclusion: The data suggest a time-dependent modulation of classical and nonclassical monocytes and chemokine receptor levels. The higher percentage of classical monocytes (inflammatory cells) suggest a residual inflammatory risk, even under preconized treatments to AMI. Indeed, these changes do not seem to be affected by choice of the lipid-lowering strategy.

Keywords: acute myocardial infarction, chemokine receptors, lipid-lowering treatment, monocyte subtypes

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Authors: Zélia Maria Da Costa Ludwig, Maria José Valenzuela Bell, Geraldo Henriques Da Silva, Thales Alves Faraco, Victor Rocha Da Silva, Daniel Rotmeister Teixeira, Vírgilio De Carvalho Dos Anjos, Valdemir Ludwig

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Advances in telecommunications grow with the development of optical amplifiers based on rare earth ions. The focus has been concentrated in silicate glasses although their amplified spontaneous emission is limited to a few tens of nanometers (~ 40nm). Recently, phosphate glasses have received great attention due to their potential application in optical data transmission, detection, sensors and laser detector, waveguide and optical fibers, besides its excellent physical properties such as high thermal expansion coefficients and low melting temperature. Compared with the silica glasses, phosphate glasses provide different optical properties such as, large transmission window of infrared, and good density. Research on the improvement of physical and chemical durability of phosphate glass by addition of heavy metals oxides in P2O5 has been performed. The addition of Na2O further improves the solubility of rare earths, while increasing the Al2O3 links in the P2O5 tetrahedral results in increased durability and aqueous transition temperature and a decrease of the coefficient of thermal expansion. This work describes the structural and spectroscopic characterization of a phosphate glass matrix doped with different Er (Erbium) concentrations. The phosphate glasses containing Er3+ ions have been prepared by melt technique. A study of the optical absorption, luminescence and lifetime was conducted in order to characterize the infrared emission of Er3+ ions at 1540 nm, due to the radiative transition 4I13/2 → 4I15/2. Our results indicate that the present glass is a quite good matrix for Er3+ ions, and the quantum efficiency of the 1540 nm emission was high. A quenching mechanism for the mentioned luminescence was not observed up to 2,0 mol% of Er concentration. The Judd-Ofelt parameters, radiative lifetime and quantum efficiency have been determined in order to evaluate the potential of Er3+ ions in new phosphate glass. The parameters follow the trend as Ω2 > Ω4 > Ω6. It is well known that the parameter Ω2 is an indication of the dominant covalent nature and/or structural changes in the vicinity of the ion (short range effects), while Ω4 and Ω6 intensity parameters are long range parameters that can be related to the bulk properties such as viscosity and rigidity of the glass. From the PL measurements, no red or green upconversion was measured when pumping the samples with laser excitation at 980 nm. As future prospects: Synthesize this glass system with silver in order to determine the influence of silver nanoparticles on the Er3+ ions.

Keywords: phosphate glass, erbium, luminescence, glass system

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Authors: Jazim Sohail, Filipe Teixeira-Dias

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Mild traumatic brain injury (mTBI) within non-helmeted contact sports is a growing concern due to the serious risk of potential injury. Extensive research is being conducted looking into head kinematics in non-helmeted contact sports utilizing instrumented mouthguards that allow researchers to record accelerations and velocities of the head during and after an impact. This does not, however, allow the location of the impact on the head, and its magnitude and orientation, to be determined. This research proposes and validates two methods to quantify impact locations from instrumented mouthguard kinematic data, one using rigid body dynamics, the other utilizing machine learning. The rigid body dynamics technique focuses on establishing and matching moments from Euler’s and torque equations in order to find the impact location on the head. The methodology is validated with impact data collected from a lab test with the dummy head fitted with an instrumented mouthguard. Additionally, a Hybrid III Dummy head finite element model was utilized to create synthetic kinematic data sets for impacts from varying locations to validate the impact location algorithm. The algorithm calculates accurate impact locations; however, it will require preprocessing of live data, which is currently being done by cross-referencing data timestamps to video footage. The machine learning technique focuses on eliminating the preprocessing aspect by establishing trends within time-series signals from instrumented mouthguards to determine the impact location on the head. An unsupervised learning technique is used to cluster together impacts within similar regions from an entire time-series signal. The kinematic signals established from mouthguards are converted to the frequency domain before using a clustering algorithm to cluster together similar signals within a time series that may span the length of a game. Impacts are clustered within predetermined location bins. The same Hybrid III Dummy finite element model is used to create impacts that closely replicate on-field impacts in order to create synthetic time-series datasets consisting of impacts in varying locations. These time-series data sets are used to validate the machine learning technique. The rigid body dynamics technique provides a good method to establish accurate impact location of impact signals that have already been labeled as true impacts and filtered out of the entire time series. However, the machine learning technique provides a method that can be implemented with long time series signal data but will provide impact location within predetermined regions on the head. Additionally, the machine learning technique can be used to eliminate false impacts captured by sensors saving additional time for data scientists using instrumented mouthguard kinematic data as validating true impacts with video footage would not be required.

Keywords: head impacts, impact location, instrumented mouthguard, machine learning, mTBI

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Authors: Carla Teixeira, Diana Azevedo, Jonatas Bessa, Maria Guilam

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The flipped classroom is a blended learning modality that combines face-to-face and virtual activities of self-learning, mediated by digital information and communication technologies, which reverses traditional teaching approaches and presents, as a presupposition, the previous study of contents by students. In the following face-to-face activities, the contents are discussed, producing active learning. This work aims to describe the systematization process of the use of flipped classrooms as a method to develop complementary national activities in PROFSAÚDE, a professional master's program in the area of public health, offered as a distance learning course, in the network, in Brazil. The complementary national activities were organized with the objective of strengthening and qualifying students´ learning process. The network gathers twenty-two public institutions of higher education in the country. Its national coordination conducted a survey to detect complementary educational needs, supposed to improve the formative process and align important content sums for the program nationally. The activities were organized both asynchronously, making study materials available in Google classrooms, and synchronously in a tele presential way, organized on virtual platforms to reach the largest number of students in the country. The asynchronous activities allowed each student to study at their own pace and the synchronous activities were intended for deepening and reflecting on the themes. The national team identified some professors' areas of expertise, who were contacted for the production of audiovisual content such as video classes and podcasts, guidance for supporting bibliographic materials and also to conduct synchronous activities together with the technical team. The contents posted in the virtual classroom were organized by modules and made available before the synchronous meeting; these modules, in turn, contain “pills of experience” that correspond to reports of teachers' experiences in relation to the different themes. In addition, activity was proposed, with questions aimed to expose doubts about the contents and a learning challenge, as a practical exercise. Synchronous activities are built with different invited teachers, based on the participants 'discussions, and are the forum where teachers can answer students' questions, providing feedback on the learning process. At the end of each complementary activity, an evaluation questionnaire is available. The responses analyses show that this institutional network experience, as pedagogical innovation, provides important tools to support teaching and research due to its potential in the participatory construction of learning, optimization of resources, the democratization of knowledge and sharing and strengthening of practical experiences on the network. One of its relevant aspects was the thematic diversity addressed through this method.

Keywords: active learning, flipped classroom, network education experience, pedagogic innovation

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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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Authors: Vanessa G. P. Severino, Eliangela Cristina Candida Costa, Nubia Alves Mariano Teixeira Pires Gomides, Lucilia Kato, Afif Felix Monteiro, Maria Anita Lemos Vasconcelos Ambrosio, Carlos Henrique Gomes Martins

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One of the biomes present in Brazil is known as Cerrado, which is a vast tropical savanna ecoregion, particularly in the states of Goiás, Mato Grosso do Sul, Mato Grosso, Tocantins and Minas Gerais. Many species of plants are characterized as endemic and they have therapeutic value for a large part of the population, especially to the rural communities. Given that, the southeastern region of the state of Goiás contains about 21 rural communities, which present a form of organization based on the use of natural resources available. One of these rural communities is named of Coqueiros, where the knowledge about the medicinal plants was very important to this research. Thus, this study focuses on the ethnobotanical survey of this community on the use of Kielmeyera coriacea to treat diseases. From the 37 members interviewed, 76% indicated this species for the treatment of intestinal infection, leukemia, anemia, gastritis, gum pain, toothache, cavity, arthritis, arthrosis, healing, vermifuge, rheumatism, antibiotic, skin problems, mycoses and all kinds of infections. The medicinal properties attributed during the interviews were framed in the body system (disease categories), adapted from ICD 10; thus, 20 indications of use were obtained, among five body systems. Therefore, the root of this species was select to chemical and biological (antioxidant and antimicrobial) studies. From the liquid-liquid extraction of ethanolic extract of root (EER), the hexane (FH), ethyl acetate (FAE), and hydro alcoholic (FHA) fractions were obtained. The chemical profile study of these fractions was performed by LC-MS, identifying major compounds such as δ-tocotrienol, prenylated acylphoroglucinol, 2-hydroxy-1-methoxyxanthone and quercitrin. EER, FH, FAE and FHA were submitted to biological tests. FHA presented the best antioxidant action (EC50 201.53 μg mL-1). EER inhibited the bacterial growth of Streptococcus pyogenes and Pseudomonas aeruginosa, microorganisms associated with rheumatism, at Minimum Inhibitory Concentration (MIC) of 6.25 μg mL-1. In addition, the FH-10 subfraction, obtained from FH fractionation, presented MIC of 1.56 μg mL-1 against S. pneumoniae; EER also inhibited the fungus Candida glabrata (MIC 7.81 μg mL- 1). The FAE-4.7.3 fraction, from the fractionation of FAE, presented MIC of 200 μg mL-1 against Lactobacillus casei, which is one of the causes of caries and oral infections. By the correlation of the chemical and biological data, it is possible to note that the FAE-4.7.3 and FH-10 are constituted 4-hydroxy-2,3-methylenedioxy xanthone, 3-hydroxy-1,2-dimethoxy xanthone, lupeol, prenylated acylphoroglucinol and quercitrin, which could be associated with the biological potential found. Therefore, this study provides an important basis for further investigations regarding the compounds present in the active fractions of K. coriacea, which will permit the establishment of a correlation between ethnobotanical survey and bioactivity.

Keywords: biological activity, ethnobotanical survey, Kielmeyera coriacea Mart., LC-MS profile

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Authors: Carla M. Machado, André A. Silva, Armando Bastos, Telmo G. Santos, J. Pamies Teixeira

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Advanced high-strength steels (AHSS) are increasingly being used in automotive components. The use of AHSS sheets plays an important role in reducing weight, as well as increasing the resistance to impact in vehicle components. However, the large-scale use of these sheets becomes more difficult due to the limitations during the forming process. Such limitations are due to the elastically driven change of shape of a metal sheet during unloading and following forming, known as the springback effect. As the magnitude of the springback tends to increase with the strength of the material, it is among the most worrisome problems in the use of AHSS steels. The prediction of strain hardening, especially under non-proportional loading conditions, is very limited due to the lack of constitutive models and mainly due to very limited experimental tests. It is very clear from the literature that in experimental terms there is not much work to evaluate deformation behavior under real conditions, which implies a very limited and scarce development of mathematical models for these conditions. The Bauschinger effect is also fundamental to the difference between kinematic and isotropic hardening models used to predict springback in sheet metal forming. It is of major importance to deepen the phenomenological knowledge of the mechanical and microstructural behavior of the materials, in order to be able to reproduce with high fidelity the behavior of extension of the materials by means of computational simulation. For this, a multi phenomenological analysis and characterization are necessary to understand the various aspects involved in plastic deformation, namely the stress-strain relations and also the variations of electrical conductivity and magnetic permeability associated with the metallurgical changes due to plastic deformation. Aiming a complete mechanical-microstructural characterization, uniaxial tensile tests involving successive cycles of loading and unloading were performed, as well as biaxial tests such as the Erichsen test. Also, nondestructive evaluation comprising eddy currents to verify microstructural changes due to plastic deformation and ultrasonic tests to evaluate the local variations of thickness were made. The material parameters for the stable yield function and the monotonic strain hardening were obtained using uniaxial tension tests in different material directions and balanced biaxial tests. Both the decrease of the modulus of elasticity and Bauschinger effect were determined through the load-unload tensile tests. By means of the eddy currents tests, it was possible to verify changes in the magnetic permeability of the material according to the different plastically deformed areas. The ultrasonic tests were an important aid to quantify the local plastic extension. With these data, it is possible to parameterize the different models of kinematic hardening to better approximate the results obtained by simulation with the experimental results, which are fundamental for the springback prediction of the stamped parts.

Keywords: advanced high strength steel, Bauschinger effect, sheet metal forming, springback

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Authors: Denise De Fátima Fernandes Barbosa, Tyane Mayara Ferreira Oliveira, Diego Jorge Maia Lima, Paula Renata Amorim Lessa, Ana Karina Bezerra Pinheiro, Cintia Gondim Pereira Calou, Glauberto Da Silva Quirino, Hellen Lívia Oliveira Catunda, Tatiana Gomes Guedes, Nicolau Da Costa

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The aim of this study is to evaluate the use of Digital Cervicography (DC) in the diagnosis of precancerous lesions related to Human Papillomavirus (HPV). Cross-sectional study with a quantitative approach, of evaluative type, held in a health unit linked to the Pro Dean of Extension of the Federal University of Ceará, in the period of July to August 2015 with a sample of 33 women. Data collecting was conducted through interviews with enforcement tool. Franco (2005) standardized the technique used for DC. Polymerase Chain Reaction (PCR) was performed to identify high-risk HPV genotypes. DC were evaluated and classified by 3 judges. The results of DC and PCR were classified as positive, negative or inconclusive. The data of the collecting instruments were compiled and analyzed by the software Statistical Package for Social Sciences (SPSS) with descriptive statistics and cross-references. Sociodemographic, sexual and reproductive variables were analyzed through absolute frequencies (N) and their respective percentage (%). Kappa coefficient (κ) was applied to determine the existence of agreement between the DC of reports among evaluators with PCR and also among the judges about the DC results. The Pearson's chi-square test was used for analysis of sociodemographic, sexual and reproductive variables with the PCR reports. It was considered statistically significant (p<0.05). Ethical aspects of research involving human beings were respected, according to 466/2012 Resolution. Regarding the socio-demographic profile, the most prevalent ages and equally were those belonging to the groups 21-30 and 41-50 years old (24.2%). The brown color was reported in excess (84.8%) and 96.9% out of them had completed primary and secondary school or studying. 51.5% were married, 72.7% Catholic, 54.5% employed and 48.5% with income between one and two minimum wages. As for the sexual and reproductive characteristics, prevailed heterosexual (93.9%) who did not use condoms during sexual intercourse (72.7%). 51.5% had a previous history of Sexually Transmitted Infection (STI), and HPV the most prevalent STI (76.5%). 57.6% did not use contraception, 78.8% underwent examination Cancer Prevention Uterus (PCCU) with shorter time interval or equal to one year, 72.7% had no cases of Cervical Cancer in the family, 63.6% were multiparous and 97% were not vaccinated against HPV. DC identified good level of agreement between raters (κ=0.542), had a specificity of 77.8% and sensitivity of 25% when compared their results with PCR. Only the variable race showed a statistically significant association with CRP (p=0.042). DC had 100% acceptance amongst women in the sample, revealing the possibility of other experiments in using this method so that it proves as a viable technique. The DC positivity criteria were developed by nurses and these professionals also perform PCCU in Brazil, which means that DC can be an important complementary diagnostic method for the appreciation of these professional’s quality of examinations.

Keywords: gynecological examination, human papillomavirus, nursing, papillomavirus infections, uterine lasmsneop

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Authors: Robert Miller, Fred Soltero, Sandra Allan, Denise Bonilla

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The Tropical Bont Tick (TBT), Amblyomma variegatum, was introduced to the Caribbean in the mid-1700s. Since it has spread throughout the Caribbean dispersed by cattle egrets (Bubulcus ibis). Tropical Bont Ticks vector many pathogens to livestock and humans. However, only the livestock diseases heartwater, Ehrlichia (Cowdria) ruminantium, and dermatophilosis, Dermatophilus congolensis, are associated with TBT in the Caribbean. African tick bite fever (Rickettsia africae) is widespread in Caribbean TBT but human cases are rare. The Caribbean Amblyomma Programme (CAP) was an effort led by the Food and Agricultural Organization to eradicate TBTs from participating islands. This 10-year effort successfully eradicated TBT from many islands. However, most are reinfested since its termination. Pheromone technology has been developed to aid in TBT control. Although not part of the CAP treatment scheme, this research established that pheromones in combination with pesticide greatly improves treatment efficiencies. Additionally, pheromone combined with CO₂ traps greatly improves active surveillance success. St. Croix has a history of TBT outbreaks. Passive surveillance detected outbreaks in 2016 and in May of 2021. Surveillance efforts are underway to determine the extent of TBT on St Croix. Puerto Rico is the next island in the archipelago and is at a greater risk of re-infestation due to active outbreaks in St Croix. Tropical Bont Ticks were last detected in Puerto Rico in the 1980s. The infestation started on the small Puerto Rican island of Vieques, the closest landmass to St Croix, and spread to the main island through cattle movements. This infestation was eradicated with the help of the Tropical Cattle Tick (TCT), Rhipicephalus (Boophilus) microplus, eradication program. At the time, large percentages of Puerto Rican cattle were treated for ticks along with the necessary material and manpower mobilized for the effort. Therefore, a shift of focus from the TCT to TBT prevented its establishment in Puerto Rico. Currently, no large-scale treatment of TCTs occurs in Puerto Rico. Therefore, the risk of TBT establishment is now greater than it was in the 1980s. From Puerto Rico, the risk of TBT movement to the American continent increases significantly. The establishment of TBTs in the Americas would cause $1.2 billion USD in losses to the livestock industry per year. The USDA Agricultural Research Service recently worked with the USDA Animal Health Inspection Service and the Puerto Rican Department of Agriculture to modernize the management of the TCT. This modernized program uses safer pesticides and has successfully been used to eradicate pesticide-susceptible and -resistant ticks throughout the island. The objective of this work is to prevent the infestation of Puerto Rico by TBTs by combining the current TCT management efforts with TBT surveillance in Vieques. The combined effort is designed to eradicate TCT from Vieques while using the treated cattle as trap animals for TBT using pheromone impregnated tail tags attached to treated animals. Additionally, active surveillance using CO₂-baited traps combined with pheromone will be used to actively survey the environment for free-living TBT. Knowledge gained will inform TBT control efforts in St. Croix.

Keywords: Amblyomma variegatum, caribbean, eradication, Rhipicephalus (boophilus) microplus, pheromone

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Authors: Pedro M. Rodrigues, Claudia Raposo, Denise Schrama, Marco Cerqueira

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Farmed fish welfare is a very recent concept, widely discussed among the scientific community. Consumers’ interest regarding farmed animal welfare standards has significantly increased in the last years posing a huge challenge to producers in order to maintain an equilibrium between good welfare principles and productivity, while simultaneously achieve public acceptance. The major bottleneck of standard aquaculture is to impair considerably fish welfare throughout the production cycle and with this, the quality of fish protein. Welfare assessment in farmed fish is undertaken through the evaluation of fish stress responses. Primary and secondary stress responses include release of cortisol and glucose and lactate to the blood stream, respectively, which are currently the most commonly used indicators of stress exposure. However, the reliability of these indicators is highly dubious, due to a high variability of fish responses to an acute stress and the adaptation of the animal to a repetitive chronic stress. Our objective is to use comparative proteomics to identify and validate a fingerprint of proteins that can present an more reliable alternative to the already established welfare indicators. In this way, the culture conditions will improve and there will be a higher perception of mechanisms and metabolic pathway involved in the produced organism’s welfare. Due to its high economical importance in Portuguese aquaculture Gilthead seabream will be the elected species for this study. Protein extracts from Gilthead Seabream fish muscle, liver and plasma, reared for a 3 month period under optimized culture conditions (control) and induced stress conditions (Handling, high densities, and Hipoxia) are collected and used to identify a putative fish welfare protein markers fingerprint using a proteomics approach. Three tanks per condition and 3 biological replicates per tank are used for each analisys. Briefly, proteins from target tissue/fluid are extracted using standard established protocols. Protein extracts are then separated using 2D-DIGE (Difference gel electrophoresis). Proteins differentially expressed between control and induced stress conditions will be identified by mass spectrometry (LC-Ms/Ms) using NCBInr (taxonomic level - Actinopterygii) databank and Mascot search engine. The statistical analysis is performed using the R software environment, having used a one-tailed Mann-Whitney U-test (p < 0.05) to assess which proteins were differentially expressed in a statistically significant way. Validation of these proteins will be done by comparison of the RT-qPCR (Quantitative reverse transcription polymerase chain reaction) expressed genes pattern with the proteomic profile. Cortisol, glucose, and lactate are also measured in order to confirm or refute the reliability of these indicators. The identified liver proteins under handling and high densities induced stress conditions are responsible and involved in several metabolic pathways like primary metabolism (i.e. glycolysis, gluconeogenesis), ammonia metabolism, cytoskeleton proteins, signalizing proteins, lipid transport. Validition of these proteins as well as identical analysis in muscle and plasma are underway. Proteomics is a promising high-throughput technique that can be successfully applied to identify putative welfare protein biomarkers in farmed fish.

Keywords: aquaculture, fish welfare, proteomics, welfare biomarkers

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Authors: Patricia Shirley Almeida Prado, Selma Paixão Argollo, Maria De Fátima Teixeira Guimarães, Leticia Matos Sobrinho

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Skeletal differential diagnosis is essential in forensic anthropology in order to differentiate skeletal trauma from normal osseous variation and pathological processes. Thus, part of forensic anthropological field is differentiate skeletal criminal injuries from the normal skeletal variation (bone fusion or nonunion, transitional vertebrae and other non-metric traits), non-traumatic skeletal pathology (myositis ossificans, arthritis, bone metastasis, osteomyelitis) from traumatic skeletal pathology (myositis ossificans traumatic) avoiding misdiagnosis. This case shows the importance of effective pathological diagnosis in order to accelerate the identification process of skeletonized human remains. THE CASE: An unidentified skeletal remains at the medico legal institute Nina Rodrigues-Salvador, of a male young adult (29 to 40 years estimated) showing a massive heterotopic ossification on its right tibia at upper epiphysis and adjacent articular femur surface; an extensive ossification on the right clavicle (at the sternal extremity) also presenting an heterotopic ossification at right scapulae (upper third of scapulae lateral margin and infraglenoid tubercule) and at the head of right humerus at the shoulder joint area. Curiously, this case also shows an unusual porosity in certain vertebrae´s body and in some tarsal and carpal bones. Likewise, his left fifth metacarpal bones (right and left) showed a healed fracture which led both bones distorted. Based on identification, of pathological conditions in human skeletal remains literature and protocols these alterations can be misdiagnosed and this skeleton may present more than one pathological process. The anthropological forensic lab at Medico-legal Institute Nina Rodrigues in Salvador (Brazil) adopts international protocols to ancestry, sex, age and stature estimations, also implemented well-established conventions to identify pathological disease and skeletal alterations. The most compatible diagnosis for this case is hematogenous osteomyelitis due to following findings: 1: the healed fracture pattern at the clavicle showing a cloaca which is a pathognomonic for osteomyelitis; 2: the metacarpals healed fracture does not present cloaca although they developed a periosteal formation. 3: the superior articular surface of the right tibia shows an extensive inflammatory healing process that extends to adjacent femur articular surface showing some cloaca at tibia bone disease. 4: the uncommon porosities may result from hematogenous infectious process. The fractures probably have occurred in a different moments based on the healing process; the tibia injury is more extensive and has not been reorganized, while metacarpals and clavicle fracture is properly healed. We suggest that the clavicle and tibia´s fractures were infected by an existing infectious disease (syphilis, tuberculosis, brucellosis) or an existing syndrome (Gorham’s disease), which led to the development of osteomyelitis. This hypothesis is supported by the fact that different bones are affected in diverse levels. Like the metacarpals that do not show the cloaca, but then a periosteal new bone formation; then the unusual porosities do not show a classical osteoarthritic processes findings as the marginal osteophyte, pitting and new bone formation, they just show an erosive process without bone formation or osteophyte. To confirm and prove our hypothesis we are working on different clinical approaches like DNA, histopathology and other image exams to find the correct diagnostic.

Keywords: bone disease, forensic anthropology, hematogenous osteomyelitis, human identification, human remains

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