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Item Métodos diagnósticos para o vírus mayaro: revisão sistemática e avaliação molecular em pacientes arbovirose like em unidade municipal de sáude de Goiânia-Goiás(Universidade Federal de Goiás, 2020-02-17) Abrantes, Gabrielly Regis; Anunciaçāo, Carlos Eduardo; http://lattes.cnpq.br/4354412874919580; Lacerda, Elisângela de Paula Silveira; http://lattes.cnpq.br/9390789693192751; Lacerda, Elisângela de Paula Silveira; Anunciaçāo, Carlos Eduardo; Lino, Guilherme; Mrué, FátimaDiseases caused by viruses are considered an often neglected public health problem that affects thousands of people worldwide, every year. Symptoms like fever, arthralgia and rash. classify patients as syndromic dengue or arbovirus like. The differential diagnosis of arbovoresis is by serological and molecular techniques. The present study aimed to evaluate diagnostic techniques in studies published in the literature in order to develop a Systematic Literature Review on the detection of the Mayaro virus that allows health professionals to carry out an efficient laboratory diagnosis, as well as the application of molecular tests for the diagnosis of MAYV. The available literature was evaluated by systematic review, focusing on the diagnosis of the Mayaro virus in humans and its methodologies. The systematic review was carried out through an exploratory study in the databases: Scielo (Scientific Eletronic Library Online), Medline (Medical Literature Analysis and Retrieval System Online), Scopus database, PubMed, LILACS from June 8, 2019 to 02 November 2019, selecting scientific articles according to the criteria of the PRISMA (2009) recommendation. The data showed that the circulation of the Mayaro virus in urban regions is already well established worldwide and that the misdiagnosis is frequent due to similar symptoms with other arboviruses. There is a wide use of varied methods over time in the diagnosis for this arbovirus, requiring further research to implement a standardized, sensitive and specific method. In a second stage of the research, between May to August 2017 and January to June 2018, 452 samples of patients with syndromic dengue symptoms (arboviruses like) were screened. The samples were sent to the Molecular Genetics and Cytogenetics Laboratory of the Federal University of Goiás, where they were subjected to molecular screening by Polymerase Chain Reaction followed by Reverse Transcriptase (RT-PCR) reaction. All positive cases for the first PCR were confirmed on RT-qPCR for viral detection. Thirteen positive samples went through both tests and thus were subjected to Sanger sequencing. In this experimental study, 17 samples were positive for the Mayaro virus infection (RT-PCR and RT-qPCR), and co-infections with the Dengue virus were also found by molecular screening carried out in parallel for the Dengue virus by RT-PCR and RT-qPCR. The positive samples for MAYV / DENV were analyzed phylogenetically, suggesting circulation of Dengue serotype 2 and a Peruvian strain of Mayaro Virus (strain IQ 4235) in Goiânia.Item Citogenômica de pterodon pubescens e citogenética comparativa com P. emarginatus (leguminosae)(Universidade Federal de Goiás, 2020-03-05) Albernaz, Victória Borges; Souza, Luiz Gustavo Rodrigues; http://lattes.cnpq.br/3626102935973855; Soares, Thannya Nascimento; http://lattes.cnpq.br/5590256762396056; Soares, Thannya Nascimento; Harand, Andrea Pedrosa; Vianello, Rosana PereiraThe genus Pterodon Vogel (Leguminosae) has only four species, of which P. pubescens (Benth.) Benth. and P. emarginatus Vogel (both known as "white sucupira") are the closest phylogenetically. These species have a wide geographical distribution in Brazil and have chromosome number 2n = 16 with small and morphologically similar chromosomes. The objective of the present work was to carry out a comparative analysis of the genome size, the banding pattern and the composition of repetitive elements in the chromosomes of P. pubescens and P. emarginatus, aiming to enhance the cytogenomic and evolutionary knowledge of these species. For this, cytogenetic characterization was performed by analyzing the number and chromosomal morphology, CMA and DAPI banding, hybridization with DNAr 5S and 35S and determining the genome size by flow cytometry. In addition, we used P. pubescens genome sequencing by NGS (Next Generation Sequencing) using the Illumina platform to characterize repetitive genomic fractions, using a Galaxy/RepeatExplorer-Elixir platform. The most abundant elements of the P. pubescens genome were located on the chromosomes by fluorescent in situ hybridization (FISH) and transferred to P. emarginatus. The species showed very similar karyotypes with: (i) CMA+/DAPI- bands in the terminal region of two chromosome pairs and pericentromeric region of all chromosomes; (ii) two pairs of 35S rDNA sites co-located with the terminal CMA+ bands; (iii) a pair of 5S rDNA sites located in the proximal region of a chromosomal pair. The genome size of P. pubescens and P. emarginatus was also similar, 1C = 0.665 pg and 1C = 0.620 pg, respectively. The repetitive fraction represented 26,4% of the P. pubescens sequenced genome, with Ty3-Athila Ty3-Athila (24,24%), Ty3-Tekay (21,93%) and Ty1-Ale (3,37%) retrotransposons being the most abundant elements. Low abundance satellite DNAs were identified: PubSat1-254 (2,09%), PubSat2-76 (2,06%), PubSat3-216 (0,58%), PubSat4-138 (0,23%). In situ hybridization reveal that all analyzed repeats were enriched in proximal CMA+ heterochromatin in both species, except for the Ty1-Ale retroelement, which was also dispersed also in the euchromatin of P. pubescens chromosomes. The cytomolecular similarity observed here suggests that the genomes of P. pubescens and P. emarginatus have highly similar repetitive fractions, which corroborates their phylogenetic proximity. However, the recent expansion of the Ty1-Ale element in the P. pubescens genome suggests some degree of differentiation in the repetitive fractions of these genomes.Item Identificação e caracterização de microRNAs no fungo dimórfico Histoplasma capsulatum(Universidade Federal de Goiás, 2020-03-06) Fernandes, Lucas Barros; Curcio, Juliana Santana de; http://lattes.cnpq.br/9577753232871923; Soares, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637; Tomazett, Mariana Vieira; Pereira, Maristela; Soares, Célia Maria de AlmeidaMicroRNAs are small non-coding RNA molecules involved in the RNA interference phenomenon. These small RNAs contain between 18 and 25 nucleotides and are derived from precursors containing a hairpin structure. Their biogenesis and mode of action are well understood in plants and animals, but in fungi their biogenesis is still poorly understood. However, small RNAs similar to microRNAs (miRNAs-like) have been identified in various fungi, such as Neurospora crassa, Sclerotinia sclerotiorum, Penicillium marneffei, Trichoderma reesei and Paracoccidioides brasiliensis. Even though the amount of data on these molecules in the Fungi kingdom has been increasing, up to now there are no descriptions of microRNAs in important human pathogens such as Histoplasma capsulatum. Therefore, due to the absence of the characterization of this regulatory mechanism in this human pathogen, this study aimed to identify microRNAs in this pathogen and to infer the biological processes regulated by these interference RNAs in this fungus. In order to do so, the methodology employed in this study was the search for published microRNAs sequences for other fungi in the literature and in silico analysis to predict the homology of these known sequences to genome regions of two H. capsulaum strains H88 and G217B. After the bioinformatics analysis, we identified six microRNAs in H. capsulatum H88 and four microRNAs in H. capsulatum G217B .One of the microRNAs was conserved in both strains. This microRNA was selected for target investigation. We identified 551 targets in the H88 strain and 495 targets in the G217B strain. Biological processes identified as being regulated by this microRNA comprise the synthesis and the degradation of cell wall components, the energetic metabolism and DNA processing. The results imply the presence of microRNAs in this fungi and a high number of targets for each microRNA, which hints on the existence of a complex network of microRNA post-transcriptional regulation in H. capsulatum.Item Análise proteômica do fungo patogênico humano Fonsecaea pedrosoi, cultivado nas condições de temperatura de 22º C e 36º C(Universidade Federal de Goiás, 2017-09-11) Lima, Davi Vinícius de; Bailão, Alexandre Melo; http://lattes.cnpq.br/5415221996976886; Bailão, lexandre Melo; Paccez, Juliano Domiraci; Novaes, EvandroThe Polymorphic fungus Fonsecaea pedrosoi is the main etiological agent of Chromoblastomycosis (CBM), characterized as a chronic cutaneous mycotic infection, occurring mainly in tropical regions. The limited availability of effective therapeutic protocols coupled with CBM clinical polymorphism lead to often prolonged and high relapse rate therapies. Results regarding the molecular biology of F. pedrosoi are scarce however, some virulence factors have already been described, evidencing the need to explore the mechanisms used by this pathogen during infection. The present work carried out in silico analyzes under specific culture conditions, with temperatures of 22º C and 36º C, aiming to better understand the proteome of this fungus, both intracellular and secreted. In order to identify these proteins, we used bioinformatic tools that are available online and allowed the prediction of protein sets that can be secreted by classical and non-classical routes. After the in silico prediction method, proteins were identified in the fungal extracts by the Ultraperformance Liquid Chromatography coupled to Mass Spectrometry (UPLC-MSE) methodology. Those identified in the extracts that brought a more reliable interpretation corroborated the results in silico. The proteins identified stand out for being related to various biological processes, such as metabolism of carbon compounds, energy, proteins related to thermal and oxidative stress, defense, proteins related to the cell cycle, among other functions. The analyzes showed that the vast majority of identified proteins are secreted by alternative pathways, and the results point to the great importance of the biological roles that these secreted proteins can play during the onset of infection. Thus, we observed that the fungus presents variations in protein expression levels according to the condition addressed and may reflect on its behavior and responses to the successful establishment of the infection.Item Estrutura genética espacial intrapopulacional em Stryphnodendron adstringens (barbatimão fabaceae)(Universidade Federal de Goiás, 2020-04-13) Miranda, Alline Afonso do Nascimento Creado de; Telles, Mariana Pires de Campos; ttp://lattes.cnpq.br/4648436798023532; Telles, Mariana Pires de Campos; Braga, Ramilla dos Santos; Pinto, Rafael BarbosaThe Cerrado biome, despite containing one of the greatest biodiversity on the planet, undergoes constant deforestation due to the expansion of agricultural frontiers. The loss of natural territory consequently leads to the loss of genetic diversity. This is worrying when it comes to native species, such as Stryphnodendron adstringens, popularly known as barbatimão. Therefore, it is important to know the characteristics of the biology of S. adstringens, in order to define efficient management and conservation strategies. In this context, the objective of this study was to evaluate genetic diversity, genetic flow (estimating the size of the neighborhood group), the presence of clones and the spatial genetic structure existing in a natural population of S. adstringens. For this, genomic DNA was extracted from the leaf tissue of 124 individuals, all georeferenced. For the amplification of the microsatellite regions, nine pairs of primers developed for the species were used. Based on the genotype matrix, analyzes of genetic diversity, effective size of inbreeding, cross-fertilization rate, spatial genetic structure and identity analysis were performed to verify the occurrence of clones. For the nine loci, a total of 74 alleles were found and an average of 8.2 alleles per location. The observed average heterozygosity (HO) was equal to 0.556 and the average genetic diversity (HE) was equal to 0.624. The inbreeding coefficient (f) was significant and equal to 0.110, showing an excess of homozygotes. The genetic diversity in the population was considered high, with 71% of the maximum expected heterozygosity. The crossing system was considered mixed based on the cross fertilization rate (t ̂a) equal to 0.80. The effective size of the consanguinity (Nef) was 111.72; corresponding to a genetic representation of 0.90, indicating that there is no genetic deviation in the population. There was a significant spatial genetic structure in the first distance class with pairs of up to 49 meters. A Sp statistic equal to 0.0095 was considered moderate and associated with the genetic neighborhood (Nb = 105.3) demonstrated that pollen dispersion is efficient to maintain genetic diversity in the population. Ten groups of individuals with identical genotypes were found for nine sites used, suggesting the occurrence of vegetative reproduction. The results plan was possible, which, for the formation of germplasm banks, is collected at a minimum distance of 49 meters between the guaranteed items, in addition to contemplating the genetic diversity present in the population.Item Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre(Universidade Federal de Goiás, 2020-04-27) Petito, Guilherme; Soares, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637; Soares, Célia Maria de Almeida; Castro, Agenor de; Silva, Lívia do Carmo; Curcio, Juliana Santana de; Silva, Kleber Santiago Freitas eCopper is an important micronutrient that acts as a cofactor in important enzymes that participate in processes of obtaining energy, reducing reactive oxygen species and metal uptake such as iron. Studies with different pathogenic fungi demonstrate the importance of an efficient copper homeostasis system, as well as the impact of this metal deprivation on the metabolism of these organisms. However, little is known about the impact of copper deprivation on pathogenic fungi of the genus Paracoccidioides. Thus, the objective of this study was to analyze the proteomic behavior of the fungus Paracoccidioides brasiliensis during copper deprivation, identifying metabolic changes in face of this condition. Proteomic analysis was performed using mass spectrometry and labelling the samples with iTRAQ (Isobariq tag for relative and absolute quantitation). In addition, methodologies such as qRT-PCR, fluorescence microscopy, enzymatic activities and Western bloting were applied in this study in order to support the proteomic data analyzed. We identified 183 differentially expressed proteins. Proteins related to gluconeogenesis, beta-oxidation and cell wall remodeling were up-regulated. We identified decreased expression of an important detoxification enzyme, Cu / Zn dependent Superoxide Dismutase (Sod1p). On the other hand, the enzymes Thioredoxin (Trxp), Glutathione S-transferase (GST) and Mn / Fedependent superoxide dismutase (Sod2p) were up-regulated. Also, NADPH production-related to the pentose-phosphate pathway enzymes. detoxifying enzyme reducing agent. An alternative oxidase (Aoxp) was up-regulated, while enzyme activity tests showed that the activity of the enzyme Cytochrome c oxidase (Coxp) was decreased. An iron uptake-related copper chaperone, Atx1 and described as important in response to oxidative stress, was up-regulated. We propose here an overview that presents the main metabolic changes in P. brasiliensis under copper deprivation and that include changes in the energy acquisition process, carbon metabolism flow, wall modeling, detoxification and respiration processes.Item Análise proteômica comparativa entre Paracoccidioides brasiliensis e Paracoccidioides americana(Universidade Federal de Goiás, 2019-12-19) Portis, Igor Godinho; Ricart, Carlos André Ornelas; http://lattes.cnpq.br/1271167329803156; Soares, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637; Soares, Célia Maria de Almeida; Lima, Patrícia de Sousa; Curcio, Juliana Santana De; Pereira, Maristela; Rocha, Juliana Alves ParenteThe genus Paracoccidioides comprises termo dimorphic fungi that cause paracoccidioidomycosis, the most prevalent systemic mycosis in Latin America. These fungi can grow in yeast at 36 ° C and mycelium at temperatures below 28 ° C. Studies have shown that the fungus Paracoccidioides has molecular differences that allow the classification of species. Initially those fungi were classified in a single species, Paracoccidioides brasiliensis, but with the advancement of some molecular and genomic biology techniques, Paracoccidioides is currently classified into five species, P. lutzii, P. brasiliensis, P. americana, P. restrepiensis and P. venezuelensis. However, different species may have different proteomes and cellular responses. In this way in our study, we performed proteomic analyzes of two species of Paracoccidioides, P. americana and P. brasiliensis. iTRAQ labelling and mass spectrometery allowed the identification of 387 statistically significant proteins; two hundreds and two in P. brasiliensis and 111 proteins in P. americana. The identified proteins have different functions, such as amino acid metabolism, energy, cell defense, among others. The results showed differences in protein responses between P. americana and P. brasiliensis in proteins related to cell defense, mitochondria and energy metabolism, suggesting that these fungi may have different metabolism and pathways for growth, profliferation and virulence.Item Regra das Ilhas e genética quantitativa evolutiva do tamanho corporal em Elephas maximus borneensis(Universidade Federal de Goiás, 2020-03-24) Silva, Felipe Naves; Diniz Filho, José Alexandre Felizola; http://lattes.cnpq.br/0706396442417351; Diniz Filho, José Alexandre Felizola; Telles, Mariana Pires de Campos; Jardim, Lucas Lacerda Caldas ZaniniThe Islands Rule describes that there is a tendency, especially in mammals, that species that are large on the continent tend to suffer from dwarfism when isolated on an island, while those that have small size on the continent tend to suffer from gigantism. Elephas maximus borneensis is small in size compared to other elephants of the genus, and there is also controversy about its condition as a natural subspecies of Borneo, when it possibly arrived on the island and the degree of isolation. For this reason, starting from the Evolutionary Quantitative Genetic Model based on the individual, we incorporate a more realistic parameterization, assuming the dioecious subspecies with sexual reproduction, balanced sexual proportion and random monogamous mating, with generations without overlapping. In addition, we use the adaptive peak on the pre-established island and also evolutionary characteristics for island populations such as: heritability, migration, inbreeding and mutation. We simulated four scenarios based on the hypotheses of colonization and evolution of the elephant's body size, taking into account the time of colonization and the possible ancestral body weight. We compared the results of the simulations with the molecular data of this subspecies and the historical records. We conclude then that among the four scenarios, only the third was supported by all the data assumed in the present study, thus, the most accepted hypothesis, describing that possibly the subspecies of Elephas maximus borneensis suffered an isolation of 11 thousand to 18 thousand years in Java and, about 700 years ago, was subjected to two subsequent founding events, arriving then on the Island of Borneo.Item Caracterização do antígeno proteico SsaA de Staphylococcus saprophyticus utilizando estratégias in silico e modelo ex vivo de infecção(Universidade Federal de Goiás, 2020-03-02) Silva, Guilherme Algusto Alves; Rocha, Juliana Alves Parente; http://lattes.cnpq.br/7089231795367245; Rocha, Juliana Alves Parente; Tomazett, Mariana Vieira; Amaral, André CorrêaStaphylococcus saprophyticus is a Gram-positive bacterium and stands out as the second pathogen responsible for diagnosed cases of urinary tract infection (UTI), affecting mainly young women. Some factors may explain the ability of S. saprophyticus to colonize periurethral, urinary and genital regions, such as the ability to bind to the epithelial tissue of the genitourinary tract and the high activity of the urease enzyme. However, few mechanisms that this bacterium uses to efficiently infect and colonize the host are fully elucidated. In species of the Staphylococcus genus, most of the known virulence factors are proteins or the pathogen's surface. Our research group identified proteins secreted from S. saprophyticus isolates that demonstrated a strong ability to stimulate the immune response in mice and one of the main immunogenic proteins identified was the Staphylococcal Antigen Secreted A (SsaA), not yet characterized in this species. In other species of the Staphylococcus genus, the SsaA protein seems to be related to virulence factors regulated by the same systems, but its specific role during infection has not yet been fully elucidated. In this sense, we propose the characterization of the SsaA protein in S. saprophyticus. Bioinformatics analyzes using the database revealed that the SsaA protein has a CHAP domain with an expected amidase function. The 3D structure of the SsaA protein was predicted through the modeling of proteins using an online server following the validation protocols, the ABCpred server was used to predict epitopes in the SsaA protein. Analysis of the phagocytosis assay revealed that blocking the SsaA protein by sera containing anti-SsaA antibody reduces the number of cells recovered thus indicating that SsaA may be important during the infectious process. Our work will contribute to elucidate the biological function of the SsaA protein, an immunogenic protein that can be useful as a diagnostic target and / or vaccine in this model.