Doutorado em Biotecnologia e Biodiversidade Rede Pró-Centro-Oeste (PRPG/UnB)

URI Permanente para esta coleção

http://200.137.215.59/tede/handle/tde/313

Navegar

Navegando Doutorado em Biotecnologia e Biodiversidade Rede Pró-Centro-Oeste (PRPG/UnB) por Por Orientador "Cruz, Aparecido Divino da"

Agora exibindo 1 - 4 de 4
  • Nenhuma Miniatura disponível
    Item
    Caracterização citogenética de bovinos da raça Nelore (Bos taurus indicus, Linnaeus 1758)
    (Universidade Federal de Goiás, 2019-09-13) Amancio, Andréia Pires; Cruz, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985; Cruz, Alex Silva da; Amaral, Alliny das Graças; Cruz, Aparecido Divino da; Silva, Claudio Carlos da; Brito, Cintia Pelegrineti Targueta de Azevedo
    The agricultural sector has been standing out in the Brazilian economy in recent decades for significant increase in productivity and its growing importance for maintaining the country's balance of trade. After the introduction of cattle in Brazil and their subsequent dispersal throughout the national territory, there was an intense process of adaptation of certain groups to specific environmental conditions of each place or region, which is responsible for the formation of several local Brazilian cattle breeds. These animals may present numerical and/or structural chromosomal alterations, which may result in a group of undesired characteristics to the producer that intends to expand his herd or even for the preservation of characteristics of a species. The aim of this study was to standardize the C, GTG and NOR banding for the bovine subspecies (Bos taurus indicus, Linnaeus 1758) and subsequently produce a pancentromeric probe by amplification and labeling the centromeric regions of autosomal chromosomes of cattle. Bovine blood samples were collected to detect metaphasic chromosomes using the cell culture method. Subsequently, cytogenetic banding, amplification, chromosomal probe labeling and FISH technique were performed. Through the banding it was possible the correct chromosomal pairing of these animals and with the use of the pancentromeric probe, the marking of the centromere regions of all autosomal chromosomes. These results may offer a new tool that meets the needs of genetic improvement of these animals.
  • Nenhuma Miniatura disponível
    Item
    Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1
    (Universidade Federal de Goiás, 2016-02-02) Gigonzac, Marc Alexandre Duarte; Pereira, Rinaldo Wellerson; http://lattes.cnpq.br/9065501029560884; Cruz, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985; Cruz , Aparecido Divino da; Ayres, Flavio Monteiro; Oliveira , Kátia Karina Verolli de; Silva, Claudio Carlos da; Reis, Angela Adamski da Silva
    X-Fragile Syndrome (FXS) is the leading cause of inherited intellectual disability in the world and the second of genetic etiology, with an estimated prevalence of 1/4000 men and 1/8000 women. The most common molecular mechanism in SXF is due to changes in the expression of the FMR1 gene, located in Xq27.3, due to CGG trinucleotide expansions in the promoter region and subsequent methylation of the gene. In spite of presenting consistent clinical findings, they are not exclusive, and the existence of carriers of alteration in the FMR1 gene without apparent clinical manifestations makes it impossible to diagnose SXF based only on the evaluation. In the present study, a methodological proposal for the molecular diagnosis of X-Fragile Syndrome was developed from the methylation-specific triple amplification of the promoter region of the FMR1 gene combined with capillary electrophoresis. Thirty-four patients with clinical indication of SXF were referred to a laboratory of the public health network. After extraction and quantification of the DNA, the samples were amplified in an optimized protocol and the products submitted to 36cm capillary electrophoresis to verify the amount of CGG repeats and the degree of DNA methylation of each sample. Pre-mutation (3%) and six complete mutations (18%) were detected, all of which revealed a high degree of methylation. Considering the clinical signs commonly presented, the patients were also analyzed for the occurrence of Autism Spectrum Disorder (ASD), which shadowing and overlapping the SXF, verifying that 100% of the individuals with complete mutation presented the phenotype. Thus, it was possible to observe small behavioral differences in the patients analyzed, indicating a lighter clinical picture regarding aspects of social interaction and stereotypies. Thus, the new methodological proposal allows to effectively determine the CGG trinucleotide expansions in FMR1 allowing an assertive diagnosis of SXF for the families of patients attended in the public health network in Goiás.
  • Nenhuma Miniatura disponível
    Item
    Aumento na taxa de diagnóstico genético dos pacientes a partir da identificação de CNVs, por CMA, envolvendo genes implicados com a manifestação clínica da deficiência intelectual
    (Universidade Federal de Goiás, 2019-05-27) Pinto, Irene Plaza; Pogue, Robert Edward; http://lattes.cnpq.br/0453496208931198; Cruz, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985; Cruz, Aparecido Divino da; Silva, Cláudio Carlos da; Minasi, Lysa Bernardes; Moura, Katia Karina Verolli de Oliveira; Brasil, Maria das Graças Nunes
    Intellectual disability (ID) is characterized by significant impairment in both cognitive and adaptive functions, originating before the age of 18 years. In addition, it is a common phenotype sign in a cluster of heterogeneous syndromic or non-syndromic disorders, associated with some comorbidities such as autism and congenital malformations. In the worldwide, ID affects around 1–3% of the general population and in Brazil ID affects approximately 0.8% of the population. The Copy number variations account for about 15– 20% of children with unexplained ID, compromising the functioning of several genes, with more than 1,416 genes described as causative of this phenotype sign. The aim of the study was to evaluate the occurrence of CNVs, identified by CMA with the size filter of < 100 kb, harboring genes functionally associated with ID in patients from SUS with a clinical diagnosis of ID referred for the genetic diagnosis. During January 2013 to December 2016, GTG banding karyotype was performed in 325 patients with ID, achieve the genetic diagnostic in 57.2%, demonstrating to be an important screening approach for patients with DI. However, 42,8% of the patients showed the karyotype with no visible numerical or structural alterations. The CMA analysis with the size filter of ≥ 100 kb was performed in these patients, where it was possible to elucidate the genetic diagnose in 29.8% of the patients, demonstrating 7,1 % of the increment on the diagnostic. All the cases remained without a diagnosis were submitted to the CMA analysis with size filter of < 100 kb, where it was identified loss CNVs in regions harboring CNTNAP2, FGF13, MID1, MID2, SHANK3, IL1RAPL1, DMD, and PAK3 genes. The reduction of the size filter demonstrated an increase of 12% in the ratio of diagnosis, expanding the spectrum of CNVs identification in regions which harboring genes related to the clinical manifestation of ID. The application of both GTG banding and CMA with the size filter of ≥ 100 kb and later the size filter of < 100 kb allowed an increase in the genetic diagnosis of ID and comorbidities, giving a broad understanding of the genetic aspects related to these conditions and allowing the adequate management of families. Finally, the genetic counseling provides a better understanding of the genetic causes of ID, the familial implications of the genetic contribution and the chance of recurrence.
  • Nenhuma Miniatura disponível
    Item
    Desenvolvimento de um sistema multiplex de loci STRs autossômicos polimórficos para a identificação humana
    (Universidade Federal de Goiás, 2017-09-05) Rodovalho, Ricardo Goulart; Pereira, Rinaldo Wellerson; http://lattes.cnpq.br/9065501029560884; Cruz, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985; Cruz, Aparecido Divino da; Vieira, Thaís Cidália; Rodrigues, Flávia Melo; Moura, Kátia Karina Verolli de Oliveira; Junior, Walter Pinto
    The main scope of the current study was to develop a short tandem repeat (STR) multiplex system, made up of 22 highly informative loci, for application in forensic genetics. The system comprised of 21 polymorphic autosomal short tandem repeat loci, namely D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX, FGA, D2S441, D17S1301, D19S433, D18S853, D20S482 and D14S1434, and the amelogenin gene locus. Strategies were developed to overcome the challenges involved in creating a multiplex system. Based on the literature and available databases, STR loci were selected to obtain discriminatory markers, and followed specific criteria for this purpose. Primers were designed using the Primer3 software and the AutoDimer was used to evaluate potential interactions between them. The 21 selected STR loci were validated individually and jointly, both to assess their sensitivity and to test the efficiency of the multiplex system. Statistical analyses were based on the genetic data of 450 unrelated individuals living in the State of Goiás, thus allowing the establishment of the parameters necessary to use this system. A total of 239 alleles were detected for the 21 loci in the set, allowing for a probability of identity of 4.23 x 10-25 to be obtained. The combined power of discrimination was 0.999999999999999999999999 and the combined power of exclusion was 0.99999. Upon complete validation of the entire system, this multiplex assay was considered to be a powerful tool for application in human identification by DNA analysis.