Mestrado em Genética e Biologia Molecular (ICB)

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    Agrotóxicos e câncer: investigação genética, sociodemográfica e exposicional em agricultores goianos
    (Universidade Federal de Goiás, 2025-09-20) Lopes, Laura de Sousa; Pedroso, Thays Millena Alves; http://lattes.cnpq.br/3806066911513227; Silva, Daniela de Melo e; http://lattes.cnpq.br/9895211901348365; Silva, Daniela de Melo e; Minasi, Lysa Bernardes; Parise, Michelle Rocha; Pedroso, Thays Millena Alves
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    Caracterização de regiões genômicas que flanqueiam os locos de resistência à antracnose do feijoeiro-comum co-4 e co-5
    (Universidade Federal de Goiás, 2014-08-15) Cieslak, Jorge Freitas; Souza, Thiago Lívio Pessoa Oliveira de; Vianello, Rosana Pereira; Coelho, Gesimária Ribeiro Costa; Borba, Tereza Cristina de Oliveira; Brondani, Claudio; Vianello, Rosana Pereira
    The common bean (Phaseolus vulgaris L.) is a legume of great importance in the diet of the population, being grown both by small and big farmers throughout the year. The influence of biotic and abiotic factors causes severe losses in yield and quality of the beans. Anthracnose is considered one of the 10 most important fungal diseases in plants. Thus, it is a priority for breeding programs the development of molecular strategies that enable the acceleration and efficiency of the selection of plants resistant to common bean anthracnose. The aim of this study was to characterize markers for the presence of SSRs and SNPs in a region approximately 100kb flanking the target loci of resistance to anthracnose Co-4 and Co-5. 15 common bean genotypes were analyzed, 12 resistant and three susceptible to anthracnose. Altogether, 10 regions SSRs were identified and genotyped for the Co-4 and Co-5 loci. Of the 10 SSRs, five were polymorphic and resulted in identification of 2.8 alleles per locus, genetic diversity and genetic distance averaged 0.3343 and 0.41, respectively. The featured SSRs did not allow discrimination of resistant and susceptible genotypes. The search for genetic regions approximately 100 Kb flanking the target locus resulted in the identification of five putative disease resistance genes annotated as encoding protein kinase. The identified putative genes sequencing was conducted with 15 genotypes of common bean totaling 7,857 bp. 99 SNPs were identified from the sequence alignment, with an average of one SNP every 79 bp. Of this total, 36 were transitions, 57 transversions and six indels. The total nucleotide diversity (θ) was 0.005127. Altogether, 60 SNPs were identified in coding regions, with 22 classified as synonymous and 38 non-synonymous mutations. In addition to the analysis of SNPs, also five primers with potential for use as STS markers were identified for the characterization of putative genes for resistance to anthracnose in common bean. The results obtained in this study are an initial step in the development of molecular markers that allow the identification of associations between haplotypes and phenotypes of tolerance to anthracnose.
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    Obtenção e investigação da atividade de nanoemulsão de óleo de melaleuca em Candida albicans
    (Universidade Federal de Goiás, 2023-12-15) Aquino, Isaque de Sousa; Amaral, André Corrêa; http://lattes.cnpq.br/8801299423520104; Amaral, André Corrêa; Rocha, Viviane Lopes; Reis, Maysa Paula da Costa
    Vulvovaginal candidiasis (VVC) is associated with the genus Candida with a high frequency among women and has been the target of extensive scientific research aimed at improving treatment alternatives for this pathology, with Candida albicans being the species found in most cases, except in cases of resistance to antifungals where non-albicans species can be found, which can also hinder the action of existing drugs responsible for treating VVC. The objective of the study was to prepare a formulation containing tea tree oil and demonstrate the effectiveness of its use in nanoemulsion as an alternative treatment for VVC. The nanoemulsion was prepared using the ultrasonification technique, demonstrating a homogeneous visual appearance without sedimentation in the final preparation. The characterization of the nanoemulsion was carried out by analyzing the size and Polydispersity Index (PDI) using the Dynamic Light Scattering (DLS) process, and for the Zeta Potential results, the Electrophoretic Light Scattering procedure was carried out, all analyzed in the equipment Zetasizer, the average values obtained were: 15.74nm in diameter, 0.4 in PDI and -25.29mV in Zeta potential. The nanoemulsion stability test over a period of 30 days showed satisfactory results with good stability of the nanoparticles. The Minimum Inhibitory Concentrations (MIC) for tea tree oil showed good results in inhibiting the growth of C. albicans ATCC 10231. Toxicity tests on red blood cells demonstrated hemolysis in the initial concentrations of the nanoemulsion and positive results in subsequent dilutions.
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    Descrição do polimorfismo do CHIT1 em grupos susceptíveis à infecção pelo fungo Paracoccidioides brasiliensis
    (Universidade Federal de Goiás, 2023-12-13) Nascimento, Tiago Lemos do; Silva, Lívia do Carmo; http://lattes.cnpq.br/7092484043564604; Amaral, André Corrêa; http://lattes.cnpq.br/8801299423520104; Amaral, André Corrêa; Silva, Daniela de Melo e; Curcio, Juliana Santana de
    Fungi of the genus Paracoccidioides are the causative agents of Paracoccidioidomycosis (PCM), an infection prevalent in Latin America that mainly affects workers whose work activity is land management. Paracoccidioides spp. have their cell walls during the yeast phase, consisting mainly of chitin, a polymer formed by β-1,4- glycosidic bonds. Humans are capable of producing chitotriosidase (CHIT-1), an enzyme that has the ability to hydrolyze these bonds present in chitin. CHIT-1 is an enzyme encoded by the CHIT1 gene, with an important role in immune defense against chitin-containing pathogens, such as fungi. Polymorphism containing the duplication of 24 base pairs in exon 10 of chromosome 1 has been associated with decreased CHIT-1 production. Therefore, this study aimed to evaluate the prevalence of the 24 bp duplication polymorphism in CHIT1 in 138 individuals, divided into four groups. Group I: patients treated at the State Hospital for Tropical Diseases – Dr. Anuar Auad (HDT) in Goiânia with a confirmed diagnosis of PCM, Group II: researchers who during their research manipulated the fungus and without a confirmed diagnosis of PCM. Group III: rural workers without a confirmed diagnosis of PCM. Group IV: people without a confirmed diagnosis of PCM. The identification of the gene polymorphism was carried out using the polymerase chain reaction (PCR) technique, observing the size of the amplicons in agarose gel. The prevalence of the 24 bp duplication in exon 10 of the CHIT1 gene in the total population was 55.1% for the homozygous wild genotype, 40.6% for the heterozygous and 4.3% for the homozygous mutant genotype
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    Genoma cloroplastidial de Serjania erecta Raldk: variação no número de genes e análise de seleção de genes de plastomas da família Sapindaceae
    (Universidade Federal de Goiás, 2022-03-04) Corvalan, Leonardo Carlos Jeronimo; Diniz Filho, José Alexandre Felizola; http://lattes.cnpq.br/0706396442417351; Nunes, Rhewter; http://lattes.cnpq.br/6169806655018346; Nunes, Rhewter; Sobreiro, Mariane Brom; Dias, Renata de Oliveira
    Serjania erecta from the Sapindaceae family is a plant with medicinal properties. Studies indicate potential for use in the treatment of Alzheimer's disease, gastric diseases and anti-inflammatory use. Although, little is known about its genetic and evolutionary aspects. The goal of this study was to assemble the chloroplast genome of S. erecta, and use it in a comparative analysis with other plastomas of the Sapindaceae family. For this, we sequenced a single specimen of S. erecta from Araxá (MG - Brasil) using Illumina Miseq. The chloroplast genome was assembled using NOVOPlasty v3.2 and annotated using the CHLOROBOX platform. For comparative analysis was used eleven chloroplast genomes from different Sapindaceae family species (Acer buergerianum, Aesculus wangii, Dimocarpus longan, Dipteronia dyeriana, Dodonaea v iscosa, Eurycorymbus cavaleriei, Koelreuteria paniculata, Litchi chinensis, Pometia tomentosa, Sapindus mukorossi, Xanthoceras sorbifolium). The chloroplast genome of S. erecta has a size of 159,297 bp with 132 genes, including 87 are protein-coding genes, 37 are tRNAs and 8 rRNAs. Among twelve chloroplast genomes avalieded, S. erecta has the lowest amount of complex repeats and microsatellites. The structure and order of genes in chloroplast genomes of the order Sapindales was extremely conserved. The variation in numbers of genes was from the 132 genes to 128 genes in the Sapindaceae family. We suggest that three factors cause variation in the number of genes in the family: (1) Inverted repeat region (IR) expansion events cause the duplication of the rpl22, rps3 and rps19 genes; (2) the pseudogenization of the rps2 gene; (3) variation in the number of genes encoding tRNAS. The phylogenetic tree had well supported nodes within Sapindaceae and Serjania formed a clade with Sapindus, Litchi, Dimocarpus, and Pometia genera. Only two genomic regions (ycf1 and ndhF) showed high nucleotide diversity and no one gene is under positive selection (ka/ks > 1). The results obtained in this study provide the assembly and annotation of the chloroplast genome of S. erecta, the first annotation of a species of the genus. It also provides an idea of how chloroplast genomes evolved in the Sapindaceae family.
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    Caracterização do genoma cloroplastidial de Caryocar cuneatum (Caryocaraceae)
    (Universidade Federal de Goiás, 2023-04-28) Mendes, Millena Silva; Nunes, Rhewter; Telles, Mariana Pires de Campos; http://lattes.cnpq.br/4648436798023532; Telles, Mariana Pires de Campos; Pinto, Rafael Barbosa; Brito, Cintia Pelegrineti Targueta de Azevedo
    The Caryocaraceae family (1845) belongs to the order Malpighiales, which has 36 families, 716 genera and 16,065 species. Caryocaraceae consists of two genera Anthodiscus G. Mey. with ten species and Caryocar L. with 16 species. The present work aims to carry out the de novo assembly of the chloroplastidial genome of Caryocar cuneatum Wittm. and to carry out comparative analyzes regarding the structure and composition of the genome with other species of Malpighiales. For this, the total DNA of an individual of C. cuneatum was sequenced using the MiSeq platform (Illumina), paired-end (2x300) with the MiSeq V3 600 cycles kit. The plastome was assembled using the NOVOPlasty v3.2 program and annotated using the CHLOROBOX. For comparative analyses, chloroplast genomes of 8 species belonging to the Malpighiales order (Caryocar brasiliense, Caryocar glabrum, Lophopyxis maingayi, Drypetes indica, Aspidopterys concava, Byrsonima crassifolia, Balanops balansae, Couepia ovalifolia) were used. The chloroplast genome of C. cuneatum had a size of 165,767 bp, composed of a single copy major region of 83,968 bp, a single copy minor region of 11,854 bp, separated by two inverted repeat regions of 34,973 bp. It has 131 genes, 90 protein coding, 33 transfer RNAs and 8 ribosomal RNAs. The rpl32 gene is not present in species in C. cuneatum and in the other two species of Caryocar. The size of the plastid genome of the genus Caryocar presents itself a lot. For the other species, there was variation in size, structure and genomic composition. The InfA gene is pseudogenized in all analyzed species and was not found in A. concava. In the analysis of synonymous and non-synonymous mutations, the rpl20 gene is under neutrality and the other genes are under negative selection. For the nucleotide diversity in Caryocar genomes, it was possible to identify two hotspot regions: trnS-GCU - trnG-UCC and rbcL - atpB, which can be used as possible molecular markers for DNA barcoding. The phylogenetic tree obtained good support values for nodes in Caryocaraceae, validating the systematic position of C. cuneatum within the family and evidence of its relationship with its sister groups C. brasilense and C. glabrum. In conclusion, this study provides the first genome of C. cuneatum sequenced, information on genomic characterization and the knowledge provided here can be used for further studies and technological.
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    Mesma informação, novas aplicações: revisitando primers do gene COI em aves e melhorando a identificação por DNA barcoding
    (Universidade Federal de Goiás, 2021-03-31) Melo, Amanda Alves de; Nunes, Rhewter; http://lattes.cnpq.br/6169806655018346; Telles, Mariana Pires de Campos; http://lattes.cnpq.br/4648436798023532; Telles, Mariana Pires de Campos; Braga, Ramilla dos Santos; Targueta, Cíntia Pelegrineti
    Molecular identification of species occurs through the comparison of either morphological, biochemical or genetic characteristics. All of these areas of knowledge for delimiting species are connected and should be analyzed as a complement to fill gaps of information of each area. The process of molecular identification of species through comparison of genetic information named DNA barcode consists of analyzing nucleotide sequences of a certain region to compare the differences between species. On the majority of the animals, including birds, the cytochrome c oxidase I (COI) gene is widely used for this purpose and in most cases, it shows the ideal greater variation in its sequence among species rather than within individuals of the same species, providing elucidative information about species differentiation. Choosing primers for a DNA barcode application is a crucial step because it depends on the aim of the study and on the taxonomic group of interest. Primers in silico analysis can evaluate its physicochemical properties, the number of species reached by them and then direct the choice of the best primers for the different research objectives, saving time and money resources. Therefore, this present work aimed to answer questions related to the efficiency of the available primers for the avian COI gene and to the avian mitochondrial genome sequences available in databases: i) What is the taxonomic coverage (number of species) reached by the avian COI gene primers? Are the universal primers actually capable of covering all species?; ii) Which avian orders have the most mitochondrial genome sequences available on the databases and what is its representativeness significance within the total number of species for each order?; iii) Which primers show the ideal physicochemical properties to increase the chances of successful amplification in laboratorial experiments? and; iv) Which primer sets are the most suited to guarantee full species recovery within the avian group?
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    Malato sintase de Paracoccidioides brasiliensis é uma proteína ligada à superfície que se comporta como uma anchorless adesina
    (Universidade Federal de Goiás, 2009-05-11) Silva Neto, Benedito Rodrigues da; Pereira, Maristela; http://lattes.cnpq.br/1345781867765758; Pereira, Maristela; Soares, Célia Maria de Almeida; Lenzi, Henrique Leonel
    The pathogenic fungus Paracoccidioides brasiliensis causative of Paracoccidioidomycosis (PCM), a pulmonary mycose acquired by inhalation of fungal airborne propagules, which may disseminate to several organs and tissues leading to a severe form of the disease. Adhesion and invasion to host cells are essential steps involved in the internalization and dissemination of pathogens. Inside host, P. brasiliensis use the glyoxylate cycle for intracellular survival. Here, we provide evidence that malate synthase of P. brasiliensis (PbMLS) is localized on the cell wall, and is secreted. PbMLS was overexpressed in Escherichia coli, and polyclonal antibody against this protein was obtained. By using Confocal Laser Scanning Microscopy and Western blot analysis, PbMLS was detected in the cytoplasm and the cell wall of the yeast phase of P. brasiliensis of mother and bud yeast cells. PbMLSr and the respective polyclonal antibody produced against this protein inhibited the interaction of P. brasiliensis to in vitro cultured epithelial cells A549. These observations indicated that cell wall-associated MLS of P. brasiliensis could be mediating the binding of fungal cells, thus contributing to the adhesion of fungus to host tissues and to the dissemination of infection.
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    Variantes polimórficas em genes de detoxificação celular e suas relações com o desenvolvimento da esclerose lateral amiotrófica
    (Universidade Federal de Goiás, 2020-03-05) Santos, Kamilla de Faria; Santos, Rodrigo da Silva; http://lattes.cnpq.br/4806187026900959; Reis, Angela Adamski da Silva; http://lattes.cnpq.br/3243656364470085; Reis, Angela Adamski da Silva; Bailão, Alexandre Melo; Souza, Guilherme Rocha Lino de
    Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease caused by the degeneration of motor neurons, leading to progressive muscular atrophy. Studies suggest the relationship of the disease with environmental and genetic factors, and the involvement of mechanisms, such as oxidative stress, with neuronal degeneration. However, the body has several ways to promote cellular detoxification against harmful compounds. The Glutathione S-transferase (GST) family consists of multifunctional enzymes that act on cell detoxification and elimination of various substances, including oxidative stress products. Among the human cytosolic classes of the GST family, two isoenzymes are highlighted: GSTM1 and GSTT1, both encoded by genes of the same name, these genes have a total deletion polymorphism that results in the absence of enzymatic activity. Therefore, the objective of the present study was to evaluate the deletion polymorphisms in GSTM1 and GSTT1 genes and their association with the risk of developing ALS. A case-control study was conducted, including 101 patients diagnosed with ALS and 119 individuals without diagnosis of neurodegenerative diseases. Peripheral blood samples were collected from both groups and subjected to DNA extraction and subsequent genotyping. The polymorphisms were genotyped by the multiplex real - time PCR (qPCR) technique, with the definition of the null and present genotypes by analysis of the melting curves produced after the amplification. Clinical and demographic data were collected from medical records and questionnaires, including topics such as cigarette use, alcohol intake, age of diagnosis, physical activity practice, occupational history, among others. Among the groups analyzed, alcohol consumption was predominant in patients with ALS, with a significant difference between the case and control groups (p=0.01). However, there was no association of GSTM1 (p=0.85), GSTT1 (p=0.90) deletion polymorphisms and their possible genotypic combinations with the risk of developing ALS. The relationship of polymorphisms with the clinical and demographic profile of patients with the disease was also performed. In this analysis, there was a significant difference in the GSTM1-present genotype with the variables: environmental exposure and smoking (p = 0.02 and 0.03, respectively), in the GSTT1-present genotype with a history of neurodegenerative disease in the family (p=0.01), while the double genotype present also showed a significant difference with the family history of neurodegenerative disease (p=0.02). In this context, the results found in this study demonstrate the non-association of GSTM1 and GSTT1 deletion polymorphisms with the development of ALS.
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    Toxicidade de nanopartículas de óxido de ferro (γ-Fe2O3) funcionalizadas com citrato ao longo do desenvolvimento inicial do zebrafish (Danio rerio)
    (Universidade Federal de Goiás, 2020-01-24) Pereira, Aryelle Canedo; Gonçalves, Bruno Bastos; http://lattes.cnpq.br/3194796958722741; Rocha, Thiago Lopes; http://lattes.cnpq.br/6325937100056775; Rocha, Thiago Lopes; Rodrigues, Gisele Augusto; Franchi, Leonardo Pereira
    Iron oxide nanoparticles (NOFs) are being increasingly used in medical, environmental and technological applications. However, with its growing production and application, concerns arise about its release into the environment and its impact on human and environmental health. In this sense, the zebrafish (Danio rerio) is proving to be an excellent model system for the analysis and classification of nanomaterial toxicity. Thus, the general objective of the present study was to evaluate the toxicity of NOFs (γ-Fe2O3 NPs) functionalized with citrate throughout the initial development of zebrafish, comparing toxicity with iron ions, after static and semi-static exposure. For this purpose, the zebrafish embryos were exposed in static (without medium renewal) and semi-static (medium renewal every 24 h) exposure modes to γ-Fe2O3 NPs functionalized with citrate and iron chloride in different concentrations environmentally relevant iron (0.3, 0.6, 1.25, 2.5, 5.0 and 10 mg L-1) for 144 h, together with the control group maintained in water reconstituted in 24-well plates. The results showed that γ-Fe2O3 NPs were accumulated mainly in the chorion of the embryos and in the digestive system and liver of zebrafish larvae. Γ-Fe2O3 NPs have low toxicity compared to iron ions, indicating that the in vivo toxicity of both forms of iron is mediated by different mechanisms of action. However, NPs caused significant changes after semi-static exposure causing a decrease in heart rate, induced blood accumulation and formation of pericardial edema in zebrafish embryos, indicating its cardiotoxic effect. Regarding the exposure method, it was observed that both forms of iron induced high embryotoxicity under semi-static exposure conditions compared to static exposure, indicating that the toxicity depends on the frequency of exposure. This was the first study on the impact of the exposure condition on the toxicity of γ-Fe2O3 NPs on the development of zebrafish.
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    Hibridização in situ por fluorescência: análise histórica, inovações e perspectivas
    (Universidade Federal de Goiás, 2022-09-05) Silva, Thainá Ferreira; Pinto, Rafael Barbosa; http://lattes.cnpq.br/9944641590654242; Soares, Thannya Nascimento; http://lattes.cnpq.br/5590256762396056; Soares, Thannya Nascimento; Nunes, Rhewter; Machado, Raquel Moura
    Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that is based on the use of fluorescence-labeled probes to identify different regions in the genome. From the improvement and development of new technologies, variations of the original technique emerged making it possible to apply it to various issues, from identifying chromosomes through descriptive cytogenetics to making evolutionary inferences or conducting applied research in biotechnology and genetic improvement. Considering the great possibility of applications of the FISH technique and its variations, the present work aims to detail the methodological principles and applications, with emphasis on plants, and to investigate its use in evolutionary studies. For this, the first chapter is a manuscript for the dissemination and popularization of science that aims to discuss the various possibilities of using the FISH technique, ranging from its methodological principles to applications in different areas. From this study, it was verified that FISH can be applied to different biological materials and regions of the genome and, despite being a technique developed for more than three decades, it can generate impacts and contributions through its wide use until today. The second chapter is a systematic review, whose main objective is to summarize and critically analyze the data available in the literature on the applications of FISH for the cytogenetic study in plants, in addition to detailing and discussing the applications of this technique for evolutionary studies in plants. This study was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The search was carried out in the Scopus database and the keywords were directed to capture the breadth of the use of the FISH technique aimed at the cytogenetic study of plants. In order to investigate the applications of this technique, the articles found were grouped into different categories, called “Plant Biotechnology”, “Evolutionary Studies”, “Descriptive”, “Techniques/Methodological” and “Gene Expression”. A total of 4006 articles were selected that showed a trend of growth in the number of publications between the years 1975 - 2020. China and the USA concentrated the largest number of articles and Brazil was in 8th position. The analysis of the most used keywords in the articles made it possible to trace a small history of the technique, ranging from the beginning of indirect labeling in plant species, passing through the advances of FISH and its variations, to the use of oligo-synthetic probes today. The categories “Plant Biotechnology” and “Evolutionary Studies” represented more than 60% of the total works, with emphasis on the use of GISH and FISH techniques with repetitive DNA probes and indirect labeling. The Poaceae family stood out in the studies of "Plant Biotechnology" and Leguminosae and Asteraceae were mostly found in the category of "Evolutionary Studies", “Comparative analysis” and “Systems and cytotaxonomy.” Thus, the understanding of the technique, the synthesis and measurement of data and the evolutionary path elaborated in this study allowed the theoretical basis about the versatility of the technique and its use today. demonstrate the main contributions of FISH directed to studies with plants, highlighting the regions of research concentration, variations of the technique, main types of studies and botanical families, as well as contributing to answering different evolutionary questions.
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    Análise proteômica do fungo patogênico humano Fonsecaea pedrosoi submetido à temperatura do hospedeiro
    (Universidade Federal de Goiás, 2021-09-29) Rosa, Gabriela Danelli; Bailão, Alexandre Melo; http://lattes.cnpq.br/5415221996976886; Bailão, Alexandre Melo; Lima, Patricia de Sousa; Rocha, Thiago Lopes
    Chromoblastomycosis (CBM) is a chronic subcutaneous mycosis, very common in tropical and subtropical regions and affects many men related to rural activities. Lesions of this disease can appear in five clinical forms and treatments are difficult due to the recalcitrant nature of the disease. The main causative agent of CBM is the fungus Fonsecaea pedrosoi, which is a polymorphic, melanized fungus that presents a certain phenotypic plasticity in face of temperature variations. Thermotolerance is one of the virulence factors of virulence for this pathogen, since it survives the temperature increase, like that of the host. However, no work so far has searched to characterize the adapted cellular processes. Proteins are part of the strategies used in response to a variety of stressors, such as host temperature. In this work we mapped the intracellular proteomic profile of F.pedrosoi cultivated under different temperature conditions (28 and 37 °C) for 24 hours. It was possible to identify by means of Liquid Chromatography coupled to Mass Spectrometry (Nano UPLC-MSᴱ) a total of 486 differentially expressed proteins, 101 of which were up-regulated and 385 down-regulated. It was possible to observe that F. pedrosoi seems to respond to temperature stress by repressing central carbon metabolism pathways, such as glycolysis, tricarboxylic acid cycle (TCA) and glyoxylate cycle, cell wall biosynthesis and melanin and induced amino acid degradation enzymes and antioxidant proteins.
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    Marcadores microssatélites para os genomas nuclear e organelar de amburana cearensis (Allemão) A.C. Smith
    (Universidade Federal de Goiás, 2021-08-30) Santos, Laís Souza; Mansano, Vidal de Freitas; http://lattes.cnpq.br/5618078551561118; Soares, Thannya Nascimento; http://lattes.cnpq.br/5590256762396056; Soares, Thannya Nascimento; Telles, Mariana Pires de Campos; Vianello, Rosana Pereira
    Amburana cearensis is a tree species, popularly known in Brazil as cerejeira, widely distributed in Brazil and also in Argentina, Bolivia, Paraguay and Peru. The species is distributed in a range of intense anthropic action and classified as endangered according to the International Union for the Conservation of Nature (IUCN). The economic demand for A. cearensis as a wood source and medicinal source has boosted its predatory exploitation, representing a threat to the survival of the species. In this scenario, conservation genetics is essential for generating knowledge about the genetic diversity of the species and defining appropriate strategies for its conservation and management. The general objective of this work was to design primer pairs with high potential for the development of robust and informative microsatellite markers for the nuclear, chloroplast and mitochondrial genomes of A. cearensis. Moreover, for quickness and cost savings, nuclear microsatellite primer pairs were designed for amplification in multiplex PCR. To achieve the proposed objectives, the genome of A. cearensis was partially sequenced, using the Illumina MiSeq platform. The assembly of the genomic library was performed from the DNA of an individual of the species and the genomic sequences obtained were analyzed using FastQC, Trimmomatic and DipSPAdes software. The search for microsatellite regions was performed using the QDD program. Thirty pairs of primers for nuclear microsatellite regions were designed, organized into five sets for amplification in multiplex PCR. The primer pairs designed in this study have the potential to generate new microsatellite markers with high individual discrimination capacity and transferability to other species of the genus Amburana. These markers can be applied quickly and efficiently for different types of genetic diversity studies of A. cearensis and Amburana genus.
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    Identificação proteômica de alterações metabólicas em Paracoccidioides brasiliensis induzidas por derivado de chalcona
    (Universidade Federal de Goiás, 2021-09-30) Carvalho Júnior, Marcos Antonio Batista de; Silva, Kleber Santiago Freitas e; http://lattes.cnpq.br/3813868830071259; Pereira, Maristela; http://lattes.cnpq.br/1345781867765758; Pereira, Maristela; Curcio, Juliana Santana de; Soares, Célia Maria de Almeida
    Paracoccidioidomycosis is one of the most important systemic mycoses in Latin America. The infection occurs through the inhalation of fungal propagules belonging to Paracoccidioides genus that are present in soils, being largely associated with the contamination of rural workers, who are constantly exposed to this potentially contaminated material. Even more than a century after its discovery, the treatment of this infection with the currently available antifungal arsenal still represents a challenge, due to the long treatment time required, as well as the high toxicity of the drugs used. These questions highlight the need for research to develop and characterize new compounds with the potential to inhibit the growth of these microorganisms. In this sense, a class of molecules called chalcones has shown great versatility for demonstrating broad biological properties, including antifungal activity among them. Through a virtual screening methodology, a chalcone derivative named compound 3 was identified by our group as a promising inhibitor of Paracoccidioides spp. Thus, in order to understand the mode of action of compound 3, we applied a proteomic approach to identify the induced and repressed proteins of P. brasiliensis in the presence of compound 3. In addition, validation assays were performed on the results found. The analysis indicated that the compound can cause an imbalance in the fungal energy homeostasis by reducing the activity of the glycolytic pathway, beta-oxidation and citric acid cycle. In vitro validations also demonstrated that reactive oxygen species accumulate within the cells during exposure to compound 3, which can destabilize cellular components such as the plasma membrane. The molecular docking assay between compound 3 and the enzyme dihydropteroate synthase, which had its expression induced after the treatment, suggest that compound 3 may act as an inhibitor of this protein, impairing the folate biosynthesis that participates as a cofactor in synthesis of nucleotides and some amino acids. Therefore, these data support the efficiency of compound 3 in producing imbalances in key pathways for the P. brasiliensis' metabolic maintenance, contributing to its antifungal role.
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    Caracterização da produção científica sobre DNA barcoding no estudo de plantas
    (Universidade Federal de Goiás, 2021-04-22) Batista, Eliane Cotrim; Gonçalves, Ariany Rosa; http://lattes.cnpq.br/6401185899301616; Telles, Mariana Pires de Campos; http://lattes.cnpq.br/4648436798023532 ; Telles, Mariana Pires de Campos; Nunes, Rhewter; Braga, Ramila dos Santos
    The application of the DNA barcoding has been shown to be successful in studies of biodiversity surveys, biomonitoring of invasive species, forensic analysis to assess food security and medicinal products based on medicinal plants, monitoring the illegal trade in threatened species, studies about food ecology, conservation initiatives, etc. The present study had as main objective to raise and systematize the scientific production on the use of DNA barcoding in plants. For this purpose, a search was carried out in the Web of Science database, using as keywords combined the regions annotated in the chloroplast and nuclear genome of Arabidopsis thaliana (model species), which are used as DNA barcoding markers, associated with the expressions: “DNA barcoding” OR “barcoding” OR “DNA barcode” OR “barcode” OR “metabarcoding”. The search was carried out covering the complete period of publication of the database until 2020. The recovered files were filtered, being considered only original articles, excluding review articles, as well as those that did not correspond to the scope of the study. A total of 1,897 articles were retrieved, of which 1,073 were within the criteria’s inclusion. As the first article was published in 2005, the analysis covered the period between 2005 and 2020. The studies were published in a total of 309 sources, with a general average of publication per year equal to 5.21. Scientific production varied over the period, but always with an increasing trend. The average number of authors per document was 3.49, since each article mostly has more than one author, which explains the average number of coauthors per document (5.34). However, out of a total of 3,747 registered authors, the number of articles per author was low, with an average equal to 0.29. China stands out as the main country, according to the origin of the authors, who has contributed with works involving the application of the DNA barcoding technique in the study of plants, being also the most cited country. The most relevant authors, based on the number of publications, were the Chinese: Chen SL, Song J, Yao H, with the article by Chen et al. (2010) the second most cited globally. The journals that showed greater prominence in relation to the number of articles published, considering the subject, were respectively: Plos One, Molecular Ecology Resources and Scientific Reports. It was possible to characterize four groups of documents regarding the use / application of the technique, respectively: Description/delimitation of species (545), Forensic Genetics (275), Development/methodology (163), Ecology (90) and 75 articles were categorized in up to two different groups. The studies grouped in the most prominent category (Description/delimitation of species), range from delimiting species from complex groups to the validation of species with properties of economic interest and species already cataloged in herbariums. The application possibilities have been expanded due to the development of High Throughput Sequencing technologies, opening doors for studies applied to Ecology, Evolution and Forensic Genetics and which use in addition to DNA barcoding, the metabarcoding.
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    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 Pereira
    The 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.
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    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 Almeida
    MicroRNAs 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.
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    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 Corrêa
    Staphylococcus 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.
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    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 Zanini
    The 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.
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    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 Barbosa
    The 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.