Doutorado em Genética e Biologia Molecular (ICB)

URI Permanente para esta coleçãohttp://200.137.215.59/tede/handle/tde/206

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Investigação de alterações bioquímicas, hematológicas e imunológicas em trabalhadores rurais do sudoeste goiano ocupacionalmente expostos a agrotóxicos
(Universidade Federal de Goiás, 2025-06-26) Silva, Iasmin Ramos da; Cardoso, Ludimila Paula Vaz; http://lattes.cnpq.br/5434857923089593; Parise, Michelle Rocha; http://lattes.cnpq.br/1256067662875050; Parise, Michelle Rocha; http://lattes.cnpq.br/1256067662875050; Reis, Fabiana Gonçalves dos; http://lattes.cnpq.br/8886148971309796; Santa Rita, Ricardo de Mattos; http://lattes.cnpq.br/1471547395688624; Costa, Rafael Menezes da; http://lattes.cnpq.br/0807014533959860; Rezende, Hanstter Hallison Alves; http://lattes.cnpq.br/4982752673858886
Embargada.
Metagenômica de solos em paisagens agrícolas no Cerrado
(Universidade Federal de Goiás, 2022-11-28) Vieira, Lucas Donizetti; Collevatti, Rosane Garcia; http://lattes.cnpq.br/9979596352166630; Collevatti, Rosane Garcia; http://lattes.cnpq.br/9979596352166630; Bessa, Layara Alexandre; http://lattes.cnpq.br/3771093495005577; Vitorino, Luciana Cristina; http://lattes.cnpq.br/5992494620867841; Borges, Clayton Luiz; http://lattes.cnpq.br/8867708267053410; Dias, Renata de Oliveira; http://lattes.cnpq.br/5189684087836977
The microbial community in the soil is highly diverse and is composed mainly of bacteria and fungi with most of these taxa currently undescribed. Here we aimed to understand the effects of landscape composition on soil microbiota diversity. Specifically, we analyzed the effects of landscape and soil variables in the composition, taxonomic, phylogenetic and functional microbial diversity in Cerrado agricultural landscapes. Kraken 2 was able to classify a mean of 7,600 species with more 10 thousand OTU classification per site and Bracken reclassify it to a mean of approximately 3,800 species per site. Archaea sequences were mostly assigned to the highly diverse phylum Euryarchaeota and bacterial sequences were predominancy assigned to the phyla Proteobacteria and Actinobacteria. MG-RAST classification using the SEED subsystems database classified over 8.2 million reads per site within the 28 functional categories. Regarding alpha diversity indexes, soil and landscape variables negatively influenced Archaea and Bacteria species Shannon diversity index, Margalef richness index, MPD and MNTD phylogenetic diversities, richness and abundance. For beta diversity, we found significant interaction with landscape variables of (APSI), percentage of pasture, habitat and forest, and local variable chromium (Cr). Taken together, our results show a taxonomical profile for Archaea and Bacteria of disturbed soils caused by changes in the landscape use. Also, the high values of alpha diversity indexes contrasting with the low values of beta diversity indexes indicate changes in species richness and its heterogeneity. Lastly, APSI and Cr are the two predictive variables that better explained the diversity and richness of Archaea and Bacteria in the Cerrado agricultural landscapes. We found that intensive land use for agriculture increases the species richness and abundance of soil microorganisms while reducing their diversity.
Recursos genômicos para o gênero Simarouba Aubl. (Simaroubaceae): caracterização genômica, análises evolutivas e marcadores moleculares
(Universidade Federal de Goiás, 2024-08-29) Silva, Marla Arianne Almeida; Nunes, Rhewter; http://lattes.cnpq.br/6169806655018346; Telles, Mariana Pires de Campos; http://lattes.cnpq.br/4648436798023532; Telles, Mariana Pires de Campos; http://lattes.cnpq.br/4648436798023532; Silva Neto, Carlos de Melo e; http://lattes.cnpq.br/2681735180356106; Sobreiro, Mariane Brom; http://lattes.cnpq.br/1549706642343277; Franceschinelli, Edivani Villaron; http://lattes.cnpq.br/5417542902696888; Soares, Thannya Nascimento; http://lattes.cnpq.br/5590256762396056
Embargada.
Análise das funções de epítopos de Staphylococcus saprophyticus na interação patógeno-hospedeiro e potencial de aplicação no diagnóstico de infecções de trato-urinário
(Universidade Federal de Goiás, 2025-01-31) Silva, Guilherme Algusto Alves; Rocha, Juliana Alves Parente; http://lattes.cnpq.br/7089231795367245; Rocha, Juliana Alves Parente; Dias, Fátima Ribeiro; Campos, Tatiana Amabile de; Borges, Clayton Luiz; Rocha, Olivia Basso
Embargado
Cienciometria em Entomogenética e recuperação de DNA Humano do sistema gastrointestinal de Diptera
(Universidade Federal de Goiás, 2024-10-29) Pires, Nayara Lopes de Souza; Pinto, Irene Plaza; http://lattes.cnpq.br/0606687569347456; Rodovalho, Ricardo Goulart; http://lattes.cnpq.br/2752094762303097; Cruz, Aparecido Divino da; : http://lattes.cnpq.br/7868817504129985; Cruz, Aparecido Divino da; Silva, Daniela de Melo e; Dias, Renata de Oliveira; Minasi, Lysa Bernardes; Gigonzac, Marc Alexandre Duarte
Data presented by the Brazilian Yearbook of Public Security (2024), points to the increasing cases of rape in Brazil. Scientific research has developed tools and techniques with high investigative potential, which have helped the justice system materialize evidence. Animal models are widely used in forensics, such as estimating the post-mortem interval. Entomogenetics, which is a sub-area of forensic entomology, has a high potential in cases of DNA recovery from the immature insects pappus of flies for the elucidation of crimes. Molecular biology tools have been applied both in the recovery of victims in advanced stages of decomposition and in the identification of aggressors in cases of sexual crimes followed by death. This thesis was structured in two chapters, the first chapter aimed to recover and isolate human DNA from the contents present in the gastrointestinal region of immature insects of a fly species from the Sarcophagidae family. Thus, conventional PCR and qPCR techniques were used. In addition, the Barcode DNA technique was used to identify the species of fly. The qPCR technique proved effective in isolating and detecting human DNA and male genetic material from the gastric system of the larvae of flies from the Sarcophagidae family. This approach was innovative and was applied for the first time with the aim of detecting human target DNA in samples from the contents stored in the stomach of larvae and pupae of flies from the family studied. Using the DNA Barcode technique, the Peckia resona species was identified, providing new perspectives for the fields of taxonomy and entomology. The second chapter, write as article, aimed to carry out a scientometric analysis of all scientific productions in the field of entomogenetics. This study offered insights into the evolution of entomogenetics-related publications over the years and provided a comprehensive understanding of the use of molecular biology methodologies to recover DNA from the digestive system of insects, the number of articles, the pioneering authors who contributed to this field, and the existing gaps in knowledge. The data revealed a scarcity of research on entomogenetics, highlighting the need to promote investigations in this area by conducting studies in different countries.
Análise proteômica da parede celular de leveduras de Paracoccidioides americana e Paracoccidioides brasiliensis
(Universidade Federal de Goiás, 2021-12-21) Malaver Salamanca, Ayda Luz; Pereira, Maristela; http://lattes.cnpq.br/1345781867765758; Soares, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637; Soares, Célia Maria de Almeida; Curcio, Juliana Santana de; Rocha, Juliana Alves Parente; Gonçales, Relber Aguiar; Pigosso, Laurine Lacerda
Paracoccidioidomycosis (PCM) is the most important systemic mycosis in Latin America. PCM is endemic in Brazil; however, it is not mandatory to report, so its actual incidence and prevalence are unknown. The etiologic agents of PCM are dimorphic fungi of the genus Paracoccidioides, which in the human host transits from mycelial to yeast cells. The cell wall is the most superficial structure of the fungus and, therefore, is important during the infection stage. The cell wall is a dynamic structure that undergoes constant remodeling, to ensure pathogen adaptation to multiple stress conditions within the human host. In addition, the cell wall also plays an important role in the processes of cell division, growth, and transition, therefore, and due to the absence of this structure in human cells, the cell wall becomes an interesting target for study. The aim of the present work was to perform proteomic study of cell wall proteins that are covalently linked by disulfide bridges, in two representative species of the genus Paracoccidioides, Paracoccidioides americana and Paracoccidioides brasiliensis. Proteomic analysis allowed the identification of differentially expressed proteins between the two species. Proteins associated with the adhesion to extracellular matrix components such as enolase, GAPDH, 14-3-3 family proteins, and proteins that bind and activate plasminogen such as enolase, fructose-1, 6-biphosphate aldolase 1 and phosphoglycerate kinase were more expressed in the P. brasiliensis cell wall. In addition, heat shock response proteins such as Hsp90 co-chaperone AHA1 and Hsp STI1 were more expressed on the cell wall of P. brasiliensis; while, catalase P was more expressed in P. americana. These results can help to understand the host-parasite relationship and guide the selection of proteins that can serve as new pharmacological targets.
Montagem e análise do transcritoma do mogno africano (Khaya grandifoliola C. DC.)
(Universidade Federal de Goiás, 2019-07-25) Soares, Sabrina Delgado; Novaes, Evandro; http://lattes.cnpq.br/0568272239145336; Novaes, Evandro; Telles, Mariana Pires de Campos; Soares, Thannya Nascimento; Coelho, Alexandre Siqueira Guedes; Vianello, Rosana Pereira
Khaya grandifoliola C. DC. is a tropical species with high value wood in international markets. Despite the high interest for reforestation and timber production, virtually nothing is known about the genetic and genomic aspects of the species. With the development of next-generation sequencing platforms (NGS), the characterization of transcriptomes can be performed on a large scale and at affordable cost. In this context, the objective of the work was to discover K. grandifoliola genes by using RNA-Seq technique and to identify those differentially expressed between the leaf and the xylem K. grandifoliola tissues. Total RNA was extracted from leaf and xylem samples of four trees planted in the experimental area of the UFG. mRNA samples were sequenced on the Illumina HiSeq 2000 platform, in paired-end mode 2 × 100 bp. Sequencing resulted in a total of 857 million sequences which, after quality control, decreased to 808 million with an average size of 24 to 85 bp. Trinity program was used to reassemble the transcriptome, obtaining 116,289 transcripts for a total of 36,271 genes. Transcripts ranged from 524 to 12,031 bp with a mean of 1,366 bp and N50 of 1,657 bp. Analyses of gene expression identified 13,626 differentially expressed genes (DEGs) (FDR <0.05) between the two tissues. Of these, 5,438 had higher expression in leaves and 8,188 in xylem. A functional enrichment analysis of Gene Ontology categories among the DEGs was performed by the Blast2GO program. This analysis identified 26 GO terms enriched among the most active DEGs in xylem. Among genes up-regulated in leaves, 48 enriched terms were found. As expected, the categories identified as enriched between DGEs point to biological processes and cellular components specific of each tissue. For example, among the most active genes in leaf tissue, terms associated with thylakoids (GO:0009579), plastids (GO:0009536) and photosynthesis (GO:0015979) were identified. On the other hand, for the xylem, terms related to carbohydrate metabolism (GO:0005975), cell wall (GO:0071554) and cytoskeletal proteins (GO:0008092) were significantly enriched. To the best of our knowledge, this is the first report of large-scale gene sequences for K. grandifoliola. Differential expression analysis allowed us to identify which genes are most active in leaf and xylem. Analysis in iPath3 identified the metabolic routes activated in leaf and xylem tissues. A BlastX search against the orange proteome (Citrus sinensis) revealed the presence of 95 DEGs annotated for the enzymes of the limonoid, a secondary metabolite with medicinal properties.
Desenvolvimento e caracterização de nanopartículas mistas de quitosana e lecitina contendo melatonina para o tratamento de feridas em ratos diabéticos
(Universidade Federal de Goiás, 2020-08-19) Corrêa, Viviane Lopes Rocha; Leite, Liliana Borges de Menezes; http://lattes.cnpq.br/2012543423092393; Amaral, André Corrêa; http://lattes.cnpq.br/8801299423520104; Amaral, André Corrêa; Bocca, Anamélia Lorenzetti; Tedesco, Antônio Cláudio; Diniz, Danielle Guimarães Almeida; Miguel, Marina Pacheco
Wound healing in diabetic patients remains a worldwide problem that can cause amputations and even lead to death. This work aimed to produce melatonin-loaded lecithin-chitosan nanoparticles (MEL-NP) and to evaluate a topical formulation containing these particles for healing in an in vivo animal model for diabetes. For the production of nanoparticles, an ethanolic solution containing soybean lecithin and melatonin was dropwise added to an aqueous solution of chitosan under sonication. The nanoparticles were physical-chemical characterized and evaluated in vivo for toxicity using the Galleria mellonella model and its potential for wound healing in diabetic rats. The MEL-NP were around 160 nm in size and had a zeta potential around 25 mV. The melatonin entrapment efficiency was 27%. Our results demonstrated that treatment with MEL-NP improved wound healing demonstrated by a wound closure earlier than the other treatments evaluated. An appreciated therapeutic effect was achieved by MEL-NP in the induction of fibroblasts and angiogenic proliferation. In addition, it was accompanied by an expressive collagen deposition. Taking the observed data, the MEL-NP produced could be used in the development of promising strategies for wound healing in diabetic people.
Desenvolvimento e testes in vivo de nanopartículas de quitosana contendo insulina na cicatrização de feridas em ratos diabéticos
(Universidade Federal de Goiás, 2019-04-16) Ribeiro, Maycon Carvalho; Menezes, Liliana Borges de; http://lattes.cnpq.br/2012543423092393; Amaral, André Corrêa; http://lattes.cnpq.br/8801299423520104; Menezes, Liliana Borges de; Miguel, Marina Pacheco; Mendes, Elizabeth Pereira; Souza, Taís Andrade Dias de; Silva, Luís Antônio Dantas da
Chitosan has been studied for its ability to accelerate healing and has been tested in the therapy of difficult-to-heal lesions, such as in diabetic patients. Insulin acts by stimulating the signaling pathway for wound healing. The objective of this work was to produce chitosan nanoparticles containing insulin for the evaluation of cicatrizant activity in diabetic rats. For the formation of the nanoparticles, the ionic gelation method was used. The nanoparticles were analyzed by diameter, potential zeta polydispersity index. The degree of deacetylation of chitosan by potentiometric was determined. For the insulin- associated nanoparticles, the mean diameter was 245.9 ± 25.46 nm and zeta potential of 39.3 ± 4.88 mV and PDI of 0.463 ± 0.01. The mean degree of deacetylation found was 72.95%. The Bradford assay revealed that the nanoparticles incorporated 97.19% ± 2.18 of insulin. To evaluate the healing, 72 Wistar rats were divided in four groups: sepigel (S), sepigel with insulin (SI), empty chitosan nanoparticles (QV) and chitosan nanoparticles containing insulin (IQ). The groups were subdivided into three subgroups (n = 6) according to the histological analysis times of the wound (3rd, 7th and 14th day). The induction of diabetes occurred through the intraperitoneal application of alloxan (120mg / kg). After confirmation of the diabetes state, the animals were anesthetized and the wounds were made with an 8.0 mm punch in the dorsal region. Macroscopic and microscopic analyzes were performed. It was possible to produce chitosan nanoparticles by the ionic gelation method, with desired diameter and zeta potential and polydispersity index. No differences were found in the rate of wound retraction among the four groups. The topical use of empty or insulin-containing chitosan nanoparticles in wound healing in diabetic rats was able to stimulate inflammatory cell proliferation and angiogenesis, followed by wound maturation. Differences in wound healing data from the group treated with insulin-containing nanoparticles and from the group treated with free insulin may be related to the high stability of the
Evidência do aumento de desvios mendelianos mediante análise transgeracional de mutações germinativas em população exposta à radiação ionizante de Césio-137
(Universidade Federal de Goiás, 2022-08-12) Oliveira, Lorraynne Guimarães; Costa, Emília Oliveira Alves; http://lattes.cnpq.br/2332660855010816; Cruz, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985; Silva, Daniela de Melo e; Silva, Claudio Carlos da; Dias, Renata de Oliveira; Leito Filho, Hugo Pereira; Cruz, Aparecido Divino da
On September 13th, 1987, the largest radiological accident in urban areas occurred in Goiânia, Goiás (Brazil), resulting in human, animal, plant and environmental exposure by Cesium-137. The mutagenic effects on the germline of people exposed to ionizing radiation are of particular concern, as the risk of inherited disorders can be increased. Single Nucleotide Variants are the most common form of human genetic variation and occur in greater abundance in a non-coding region and in recent years, several technologies have been developed to identify these variants. One of these technologies is Next-Generation Sequencing, which offers greater amounts of data, shorter sequencing times and reduced costs. The objective of this work is to establish the spectrum and frequency / rate of base de novo substitutions of germ origin from the trio's new generation sequencing data, corresponding to an F1 generation child and his biological parents, accidentally exposed to high and low doses of IR of cesium-137, contributing to knowledge about the biological effects of exposure to ionizing radiation. Considering case and control, the results of 38 parents, and an offspring of 14 children born from the exposed group and 5 children from the unexposed population were included. Exposed individuals had ~39% increase in global mean DMs compared to controls. Exposed mothers had ~44% increase in global mean DMs compared to controls. The A:T>C:G mutation was the one that showed the greatest statistically significant increase in occurrence in the offspring of exposed individuals. Transition replacement rates were higher than transversions in the offspring of cases and controls, but the difference is not statistically significant. In conclusion, with the methodology and biomarkers used, it was possible to identify the origin of mutation in the parents, as well as the type of substitution and to inform which variant was mutated, it was also possible to detect the frequency of the germline mutation in DM which made it possible to retrospectively study this population exposed to IR.
Análises proteômicas de cepas de Staphylococcus saprophyticus elucidam diferentes estratégias para virulência e infecção
(Universidade Federal de Goiás, 2019-05-13) Silva, Karla Christina Sousa; Rocha, Juliana Alves Parente; http://lattes.cnpq.br/7089231795367245; Rocha, Juliana Alves Parente; Marval, Márcia Giambiagi de; Fontes, Wagner; Kipnis, André; Borges, Clayton Luiz
Staphylococcus saprophyticus is a gram positive, coagulase negative bacteria and is one of the most common etiological agent of urinary tract infections (UTI) among sexually active young females. The infection may be led mainly by sexual intercourse. Little is known about the molecular features of S. saprophyticus infection model. However, plasticity in the proteomic profile is detected in the genus Staphylococcus and can be related with infection ability. In this study, we aimed to analyze the metabolic differences among three different pathogenic strains of S. saprophyticus (ATCC 15305, 7108 and 9325) using proteomics approaches. The proteomic data revealed variations among the strains that may lead to different responses in the context of an infection. There are differences in the carbohydrate metabolism, amino acid metabolism and pathogen defense against host defenses. Thioredoxin was one of the differentially expressed proteins among strains, thiol level assays showed similar levels of reduced thiol for ATCC15305 and 9325 strains, in contrast 7108 strain showed considerably low levels of reduced thiol levels when compared to the other analyzed strains. Urease is one of the main virulence factors of S. saprophyticus. The cytoplasmatic urease concentrations varied considerably. The 9325 strain had the lowest cytoplasmatic urease activity, however when plated on ureabased agar it showed a high urease activity, which means that probably most of the urease synthesized is exported to the extracellular milieu. Another important feature is related to the purine metabolism operon, which is highly regulated on ATCC15305 and exerts an influence upon the biofilm production in this strain. These results show that, although belonging to the same species, different S. saprophyticus strains present diverse behavior in response to different contexts that the microorganism may face in the host.
Investigação genômica em pacientes com deficiência intelectual ou atraso global do desenvolvimento assistidos na rede pública de saúde do estado de Goiás
(Universidade Federal de Goiás, 2022-02-21) D’Ávila, Ana Julia da Cunha Leite; Minasi, Lysa Bernardes; http://lattes.cnpq.br/9057708164796074; Cruz, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985; Cruz, Aparecido Divino da; Silva, Claudio Carlos da; Gigonzac, Thais Cidália Vieira; Costa, Emilia Oliveira Alves; Silva, Daniela de Melo e
Intellectual Disability (ID) is a complex and heterogeneous clinical condition that affects about 1 to 3% of children and adolescents worldwide, and that can be caused by a variety of environmental and/or genetic factors. There are a variety of human genetic variants already identified for ID, from small variants to larger structural variants that affect thousands to millions of nucleotides. GTG banding karyotyping and microarray chromosome analysis (CMA) are often used to identify the genetic causes of ID, especially in cases Where the clinical evaluation indicates the syndromic form of ID. The use of next generation sequencing technologies for the diagnosis of ID has made it possible to expand the detection of new mutations and new genes associated with ID. The aim of this study was to perform total exome sequencing with gene panel analysis for intellectual disability, in patients who presented karyotype without structural and/or numerical alterations and did not present pathogenic copy number variations (CNVS) in the CMA. A retrospective cohort of 369 patients of both sexes with clinical indications of DI and/or global developmental delay was evaluated in the study. Peripheral blood samples were collected to perform cytogenetic analysis and gene sequencing. GTG banding was performed for all patients following the standard protocol. CMA was performed for patients who did not present structural and/or numerical alterations in the karyotype. Genomic DNA was isolated from patients and their biological parents in order to determine the origin of the CNVs found. Cases that were not diagnosed after performing the karyotype and CMA were referred for exome sequencing with gene panel analysis for intellectual disability, which encompassed 1,252 genes. With the karyotype, it was possible to identify chromosomal alterations in 34.7% (128/369) of the patients. CMA was performed in 83 patients who had results with no alterations in karyotype (46,XX or 46,XY) with a diagnosis rate of 21.7% (18/83). Exome sequencing for target genes was performed in 19 trios of families that had negative results in previous methodologies. Gene panel analysis of exome data identified mutations in 63.1% (12/19) of cases, of which 75% (9/12) were pathogenic variants, 8.3% (1/12) probably pathogenic and in 16.7% (2/2) VUS. Of the total variants identified, 75% (9/12) were de novo mutations and 25% (3/12) had mutations inherited from healthy parents. The most prevalent variants were missense-type mutations (66.7%) followed by nonsense-type mutations 16.7%), frameshift (8.3%) and loss CNV (8.3%). With the three methodologies applied, it was possible to identify the genetic causes of ID in 42.3% (156/369) of the patients. In conclusion, our results show different methodologies that can be used for the genetic diagnosis of ID and that exome sequencing with gene panel analysis provides an efficient diagnostic strategy when combined with clinical and laboratory screening.
Homeostase de ferro em Paracoccidioides spp.: novos alvos de estudo e estabelecimento de HSP30 como proteína ligante de hemoglobina
(Universidade Federal de Goiás, 2021-06-28) Souza, Aparecido Ferreira de; Soares, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637; Soares, Célia Maria de Almeida; Schrank, Augusto; Parente, Ana Flávia Alves; Bailão, Elisa Flávia Luiz Cardoso; Bailão, Alexandre Melo
Understanding the mechanisms that govern host-pathogen interaction is crucial for the development of new therapeutic and diagnostic approaches. Fungi of Paracoccidioides genus are the etiological agents of paracoccidioidomycosis, a systemic mycosis and, when in the host, fungi find a hostile environment that is scarce in nutrients and micronutrients, such as Fe, which is indispensable for the survival of the pathogen. Previous studies have shown that fungi of this genus, in response to Fe deprivation, are able to synthesize and capture siderophores (Fe3+ chelators), use host proteins that contain Fe as metal’s source and use a non-canonical reductive pathway for assimilation of Fe. Despite all these findings, there are still gaps that need to be filled regarding the pathogen's response to Fe deprivation. In the present work, targets that still need to be studied in this context were defined, such as the establishment of the protein functions of the fungus that contain the CFEM domain and the determination of the specificity of siderophore receptors. Additionally, it was shown that the exposure of Paracoccidioides to hemoglobin promotes changes at fungal cell wall proteome level, which is illustrated by the increased ability of the fungus to interact with macrophages. This approach allowed the bioprospecting of the hemoglobin-binding protein HSP30, which is present on the cell surface and has its expression regulated positively when the fungus is exposed to hemoglobin. It has been shown that silencing of hsp30 causes decreased growth of the fungus after exposure to hemoglobin. It is also necessary to investigate the function of HSP30 as heme oxygenase. The detailed observation of those open questions can promote the expansion of knowledge about the biology of Paracoccidioides spp. and, consequently, promote the bioprospecting of new targets for differential diagnosis and/or therapy of PCM.
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 Parente
The 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.
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 e
Copper 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.
A história evolutiva de uma perereca Sul-Americana Scinax squalirostris (Lutz, 1925) (Anura, Hylidae): um resgate do passado e consequências futuras
(Universidade Federal de Goiás, 2018-10-31) Jardim, Tatianne Piza Ferrari Abreu; Maciel, Natan Medeiros; http://lattes.cnpq.br/2116561844584292; Collevatti, Rosane Garcia; http://lattes.cnpq.br/9979596352166630; Maciel, Natan Medeiros; Silva, Daniela de Melo e; Lima, Natácia Evangelista de; Lima, Luciana Signorelli Faria; Machado, Iberê Farina
Geological events of the Neogene and the climatic fluctuations of the Quaternary played an important role in shaping the landscape and climate of South America therefore directly influencing the evolutionary history of the organisms of this area over the last million years. These changes led to the alternation between warm and humid, cold and dry periods. Such alternation dictated the dynamics of retraction and expansion of open and forest landscapes. Species associated to these environments evolved following this dynamic, which lead to alteration in genetic conformation, lineage differentiation and even speciation. As in the past, future changes inclimate can modify the landscape causing changes in the geographical distribution of species. In addition, predicted global warming may lead to a decline in genetic diversity as well as lead to extinction due to species' low ability to adapt to drastic and quick changes. In this thesis two regions of mitochondrial DNA (Cytb and 12S) and one nuclear (RAG-1) were used together with coalescing simulations, and ecological niche modelling to access the evolutionary history of a Scinax squalirostris (Lutz, 1925), a species associated to the South American grasslands. In the first chapter, we sought to understand how Neogene and Quaternary geological or climatic events, respectively, may have shaped the current disjunct distribution and the genetic diversity pattern of S. squalirostris. The populations of S. squalirostris were found to have high genetic diversity, with no sign of current gene flow, a high genetic differentiation, and a stable demographic history over time with scattered origin in southern Brazil. Coalescence events date from Pliocene-Pleistocene, with haplotype sharing among geographically distant populations, which indicates incomplete lineage sorting. The paleodistribution models suggests that S. squalirostris lineages were widely distributed during the last glacial maximum (LGM) but afterwards contracting and changing their area of occurrence. These results indicate that the current geographic distribution and genetic diversity of S. squalirostris is due to the contraction of an area widely distributed in the past, generated by the dynamics of retraction of grasslands in warmer periods due to the loss of areas suitable for their occurrence. In the second chapter, we tested the hypothesis that the current populations of S. squalirostris could represent distinct lineages with candidate species not previously described, due to the current disjunct distribution. Using molecular and morphometric data the formation of two groups was rescued. One of them consists in a candidate species to be described, which is a lineage restricted to the Central-West region of Brazil. The other one comprises of populations from the South and Southeast Brazil, Paraguay, Uruguay and Argentina. In the third chapter, ecological niche modelling, molecular techniques and simulations of genetic groups were used to verify how future climate changes could alter the genetic diversity and distribution of S. squalirostris. Through two climatic scenarios with different temperature changes to 2100 (scenario 4.5 RCP increases 1.8 ° C and stabilizes, and scenario 8.5 increases 3.7 ° C and continues to increase), ecological niche modelling analysis indicated a decrease of suitable areas in the Central-West and Southeast regions, with a displacement towards the South of Brazil entering the central region of Argentina towards more anthropized areas. Most of the Central West and Northern Southeast populations may be extinct due to the absence of climatic suitable areas for their occurrence and low genetic diversity. In addition, it was observed that Protections Areas (PAs) currently harbors a large part of the genetic diversity of S. squalirostris. Thus, PAs in areas that will be ideal for the occurrence of S. squalirostris will be able to maintain their high levels of genetic diversity, but with losses of genetic diversity in the Midwest and Southeast regions. This work indicates that future climate changes will negatively affect this species, since the appropriate areas for its occurrence will be reduced and displaced. The loss and changes in genetic clusters may lead to a possible loss of the evolutionary potential of S. squalirostris populations in responding to future climate changes, which could result in the extinction of some populations.
Avaliação do dano ao DNA pelo ensaio cometa e análise dos pontos de variação dos genes CYP2E1, CYP1A1 e OGG1 em etilistas do município de Goiânia-GO
(Universidade Federal de Goiás, 2018-05-28) Lopes, Mariana Paiva; Silva, Daniela de Melo e; http://lattes.cnpq.br/9895211901348365; Silva, Daniela de Melo e; http://lattes.cnpq.br/9895211901348365; Parise, Michelle Rocha; Nunes, Hugo Freire; Costa, Emília Oliveira Alves; Avelar, Juliana Boaventura
Ethanol is considered the most consumed drug in the world, is part of several cultures. However, chronic ethanol consumption has become a social and public health problem, since it has serious health consequences and is a risk factor for several diseases and mortality. The CYP2E1 gene encodes an enzyme that is the main constituent of the microsomal oxidation system of ethanol. The CYP1A1 gene encodes an enzyme that, like CYP2E1, acts on the oxidative biotransformation of phase I substrates of the metabolism of xenobiotics such as ethanol. In addition, genes related to DNA repair such as OGG1 are also of great importance as they are involved in protecting the genome by helping to maintain the cellular functions of organisms. Problems in the repair process can interfere with aging and promote disease development. Genetic variations in these genes result in significant differences in enzyme activity. However, there is still no clear correlation between the functional significance of these variations in both ethanol metabolism and ethylene repair. In the present study, the variability of specific regions of the CYP2E1, CYP1A1 and OGG1 genes were analyzed in samples of alcoholics from a Psychosocial Care Center (CAPS) of the State of Goiás, in the city of Goiânia, and the results were compared with the data obtained by the 1000Genomas project. A study of the association between the polymorphism of these genes and the damage to the DNA in ethanol was carried out. Sequencing reactions and the comet assay were performed. There was no relation between the data of the polymorphism found with DNA damage. However, in the analysis of the DNA damage between the case and control groups, greater damage was observed in the alcoholics in relation to the control group. The genotypic frequencies adhered to that expected by the HardyWeinberg Equilibrium for all the polymorphism in the Brazilian samples. Based on the polymorphism of the CYP2E1 gene, 15 haplotypes were inferred. Of these, only 4 haplotypes were common in all populations. While based on the two polymorphism of the CYP1A1 and OGG1 genes, 3 haplotypes were inferred for each gene, all present in the Brazilian study population.
Investigações genéticas em doenças raras: uma contribuição positiva das tecnologias genômicas
(Universidade Federal de Goiás, 2017-08-09) Reis, Fabiana Gonçalves dos; Cruz, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985; Silva, Daniela de Melo e; http://lattes.cnpq.br/9895211901348365; Silva, Daniela de Melo e; Minasi, Lysa Bernardes; Gigonzac, Thaís Cidália Vieira; Paccez, Juliano Domiraci; Costa, Emília Oliveira Alves
Neurological disorders are a group of conditions that manifest early in the development of the child, so that delayed neuropsychomotor development (ADNPM) and intellectual disability (ID) can impair cognitive, language, motor and social behavior. The etiology of ID is quite heterogeneous and prenatal, perinatal and postnatal factors are associated with an increased risk of this deficiency. However, 30 to 50% of the cases remain with the unknown etiology. In this context, the main objective of this study was to identify submicroscopic genomic alterations (<5Mb) by means of microarray chromosomal analysis (CMA) in patients with clinical indication of ADNPM and/or ID, sent by attending physicians of the state public health network of Goiás. We analyzed 149 cases of both sexes. Of the total number of patients, 47 had the diagnosis clarified using cytogenetics by G banding. Of 102 patients with an incomplete diagnosis, 72 agreed to participate in the present study and, therefore, performed the CMA. The elucidation of the diagnosis by CMA was possible in 22 patients. Among the results obtained, three rare cases were selected to compose this thesis. The first case is from a patient in whom a de novo microduplication of 0.45 Mb in the 5q35.2q35.3 region containing the NSD1 gene was identified. The effect of the dosing of this gene has been related to Sotos Syndrome and its inverted phenotype. The second case shows the molecular detection of an absent allele on the X chromosome and the presence of 28 CGG repeats in FMR1 gene in the present allele. The CMA showed that the patient had a de novo microdeletion of 4.176 Mb in the Xq27.3-q28 region that affected 34 genes, including five genes (TMEM185A, TMEM257, FMR1, IDS, and FMR2) that were directly correlated with ID phenotypes and neurological disorders. The third case is a de novo microdeletion of 1.59 Mb in the 1p32.3 region involving the DHCR24 gene, which causes a gene dosage effect influencing the activation of enzymes that cause desmosterolosis, which is a desmosterol conversion disorder in cholesterol. Thus, the results of this thesis showed the relevance of the use of the CMA technology to diagnose patients with clinical signs of ADNPM and/or ID that presented karyotype without alterations, evidencing the importance of this technology for public health.
Metabolismo do acetato entre membros do gênero paracoccidioides spp
(Universidade Federal de Goiás, 2017-04-13) Mata, Fabiana Ribeiro da; Soares, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637; Soares, Célia Maria de Almeida; Ricart, Carlos André Ornelas; Ulhoa, Cirano José; Bailão, Alexandre Melo; Brito, Wesley de Almeida
Members of the genus Paracoccidioides are known pathogens of humans that can be isolated from different infection sites. To investigate the expression rates of proteins expressed by different isolates, of the genus Paracoccidioides, including one isolate of P. lutzii (Pb01), and three isolates of P. brasiliensis (Pb03, Pb339 and PbEPM83) using sodium acetate, as carbon source, proteins quantities were determined using label-free and data independent LC-MSE. Statistical analysis provided the comparison of proteins profiles in the isolates. A total of 1160, 1211, 1280 and 1462 proteins were reproducibly identified, and relatively quantified in P. lutzii e P. brasiliensis isolates Pb03, Pb339 e PbEPM83, respectively. Notably, a total of 526, 435, 744 and 747 proteins were differentially expressed among P. lutzii and the P. brasiliensis isolates Pb03, Pb339 and PbEPM83, respectively with a fold change equal or higher than 1.5. The analysis revealed the reorganization of metabolism through the induction of proteins related to gluconeogenesis, stress response and amino acid degradation in the four isolates evaluated. The differences between the isolates were observed as follows: greater increases in the expression levels of proteins belonging to the cycle of glyoxalate, TCA and respiratory chain, ethanol production and β-oxidation were observed in the isolatesPPEPM83 and Pb01; Cyclic and cyclocyclic proteins of the methylcitrate were induced in Pb339. Proteomic profiles indicate that the four isolates reorganize the metabolism for the use of acetate as a carbon source.
Aplicações toxicogenômicas em cultura de células expostas a nanomateriais e populações humanas expostas a pesticidas
(Universidade Federal de Goiás, 2017-09-04) Franco, Fernanda Craveiro; Silva, Daniela de Melo e; http://buscacv.cnpq.br/buscacv/#/espelho?nro_id_cnpq_cp_s=9895211901348365; Parise, Michelle Rocha; Silva, Claudio Carlos da; Vieira, Thais Cidália; Costa, Emília Oliveira Alves; Silva, Daniela de Melo e
The interaction of some compounds and DNA molecule can cause genotoxic changes. The tests used to evaluate genotoxicity have evolved over the years, and they have great efficiency. Currently, several traditional methodologies are used, such as the comet assay, and some more modern, based on molecular biology. Considering the importance of detecting the effects caused by exposure to potentially genotoxic agents, this thesis presents three chapters with different approaches to this topic. The first chapter aimed at evaluating published studies on the mutagenic, genotoxic and oxidative effects caused by occupational exposure to pesticides. For this, a systematic review of articles published between the years of 2011 and 2015 was carried out, focusing on these exposure conditions. The results showed that 44% of the studies addressed only men and 44% approached men and women, and 78% used rural workers as an exposed population. The most evaluated class of pesticide was organophosphorus. The studies were separated by methodologies of analysis used and all showed some type of alteration in the populations occupationally exposed to pesticides. The second chapter, published in the journal Environmental Science and Pollution Research, considered the dengue scene in the state of Goias and techniques of mosquitoes control, and sought to understand the genotoxic effects caused by exposure of endemic agents to pesticides used against Aedes aegypti, being the first study published with this group of workers. The main pesticides used were insecticides and larvicides and an increase in DNA damage was observed in the exposed population, without influence of the lifestyle factors. In the whole transcriptome assay the differential expression of genes related to various chronic diseases was observed. Finally, the third chapter was carried out in collaboration with the Institute of Experimental Medicine of the Czech Academy of Sciences, aiming the determination of cytotoxic and genotoxic effects caused by in vitro exposure to different types of metallic nanomaterials. In nanomaterials that showed cell viability reduction (cytotoxic), an increase in genotoxicity was also identified. In addition, three of the four non-cytotoxic nanomaterials evaluated showed genotoxic effects. Studies aimed at detecting the compounds genotoxic potential are extremely important for the regulatory definition and identification of risks caused by exposure. In this study, the toxicological and genotoxic effects of different compounds and mixtures were evaluated using traditional and modern techniques, in vitro or by human biomonitoring, seeking to understand and determine the risks involved in exposure to these substances.

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