Doutorado em Biotecnologia
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Navegando Doutorado em Biotecnologia por Assunto "Biomarcadores"
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- ItemAvaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua(Universidade Federal do Espírito Santo, 2023-11-22) Có, Anna Clara Gregório; Camillo, Cláudia Malheiros Coutinho; https://orcid.org/0000-0001-9016-2668; http://lattes.cnpq.br/3184503163639480; Zeidler, Sandra Lúcia Ventorin von; https://orcid.org/0000-0002-8897-5747; http://lattes.cnpq.br/5785612863130498; https://orcid.org/0000-0002-7737-0371; http://lattes.cnpq.br/3678557620411441; Nunes, Fabio Daumas; https://orcid.org/0000-0002-7785-6785; http://lattes.cnpq.br/4909755821591847; Errera, Flavia Imbroisi Valle; https://orcid.org/0000-0002-8069-6372; http://lattes.cnpq.br/9337327437538048; Arantes, Lidia Maria Rebolho Batista; https://orcid.org/0000-0001-8230-1218; http://lattes.cnpq.br/2019308149950531; Paula, Flavia de; https://orcid.org/0000-0001-8679-2982; http://lattes.cnpq.br/7913201450663683Oral squamous cell carcinoma (OSCC) is the 16th most commonly diagnosed form of cancer globally, with a higher prevalence in the tongue compared to other areas of the oral cavity. However, the lack of effective biomarkers for diagnosis, especially for precancerous lesions, poses a limitation, as visual or histological examination cannot predict the progression of dysplastic lesions, making it difficult to determine whether they will develop into cancer or return to normal epithelium. In this context, the present research aims to investigate molecular targets that may indicate the irreversible transformation of these cells, to provide a basis for broader studies aimed at using these targets as biomarkers for early OSCC diagnosis. To achieve this goal, this experimental study addressed the evaluation of the expression of a panel of microRNAs and proteins in tumour tissues and adjacent tumour-adjacent epithelium obtained from patients with tongue squamous cell carcinoma and oral epithelium from healthy individuals. Additionally, the research explored the association between these biomarkers, seeking to determine their potential application as diagnostic biomarkers for tongue squamous cell carcinoma. A total of 75 cases of tongue squamous cell carcinoma and adjacent tumouradjacent epithelium were included in the study, and the expression of the proteins survivin, Bcl-2, PLK1, p16, p40, p63, EGFR, and cyclin D1 was analysed by immunohistochemistry. The analysis of the microRNA panel's expression involved 31 samples of tongue squamous cell carcinoma, 10 samples of healthy gingival tissue, and 10 samples of serum from healthy individuals, as well as 7 samples of serum from patients diagnosed with OSCC, using the RT-qPCR technique. In silico analysis by bioinformatics validated the findings related to the expression of differentially expressed microRNAs in the sample group. The results showed differences in the expression of miRNA-31-5p (p<0.001) and miRNA-21-5p (p=0.001) in tumour samples compared to control samples. Significant differences were not observed in the expression of miRNA-24-3p, while miRNA-542-3p and 196a-5p were not detected in the sample group. No significant difference was observed in the expression of miRNAs in serum samples. The assessment of the diagnostic potential of microRNAs included ROC curve analysis, which revealed that miR-21-5p had an area under the curve (AUC) of 0.803, while miR-31-5p obtained an AUC of 0.777. The results also identified differential expression among the proteins survivin, PLK1, and p63, all of which showed increased expression in tumour tissue. Additionally, a correlation was observed between the expression of miR-21-5p and the protein p40 (chi-square: p=0.047; Spearman correlation: r=0.402; p=0.023). In conclusion, the results suggest that miR-21-5p and miR-31-5p may be potential diagnostic biomarkers for tongue squamous cell carcinoma, providing a foundation for further exploration for large-scale studies to explore miRNA-protein correlations, considering the site specificity of miRNAs.
- ItemDeterminação do perfil molecular dos extratos de folhas de Carica papaya (L.) em busca de biomarcadores estágio-dependentes relacionados à doença da meleira do mamoeiro(Universidade Federal do Espírito Santo, 2024-07-12) Britto, Isabella Oliveira; Santos, Alexandre Martins Costa; https://orcid.org/0000-0002-8801-8875; http://lattes.cnpq.br/; https://orcid.org/; http://lattes.cnpq.br/; Leite, João Paulo Viana; https://orcid.org/; http://lattes.cnpq.br/; Silva, Diolina Moura ; https://orcid.org/; http://lattes.cnpq.br/; Gonçalves, Juliana Barbosa Coitinho; https://orcid.org/; http://lattes.cnpq.br/; Lima, Graziela Domingues de Almeida; http://lattes.cnpq.br/Papaya Sticky Disease is caused by PMeV viral complex and poses a significant threat to papaya production worldwide. Infected plants remain asymptomatic until flowering and fructification, acting as silent reservoirs of the virus in the field. Secondary metabolites could act as potential biomarkers of disease progression and can be detected using chromatographic techniques for early disease diagnosis. In this study, molecular profiles of ethanolic extracts from C. papaya leaves in pre- and postflowering (2.5, 5.0, and 7.5 mg.mL-1) and fractions were evaluated by UV-Vis spectroscopy, reversed-phase chromatography, ion exchange chromatography, and mass spectrometry. The 7.5 mg.mL-1 concentration was selected as the experimental concentration for crude extracts. Chromatograms showed symmetrical peaks, eluted within similar retention time ranges across the three plant stages, with narrow base widths, similar peak shapes, and no tailing, suggesting the presence of a limited number of potentially isolable compounds. Analytical parameters indicated quantitatively larger chromatographic peaks in pre-flowering leaf extracts compared to post-flowering ones, reflecting a possible defense response against viral infection. Significant differences in chromatographic profiles between the different plant stages were observed, indicating the potential of these groups as biomarkers for Papaya Sticky Disease. This study developed a sensitive and reproducible chromatographic method to distinguish plant stages, identifying flavonoids and alkaloids as components of strategic groups related to the defense system against viral infections.
- ItemValor prognóstico do infiltrado linfocitário tumoral e da expressão de PLK1 e FOXM1 em carcinoma epidermoide oral(Universidade Federal do Espírito Santo, 2022-07-25) Damasceno, Thabata Coeli Dias; Zeidler, Sandra Lucia Ventorin Von; https://orcid.org/0000000288975747; http://lattes.cnpq.br/5785612863130498; https://orcid.org/0000000244434025; http://lattes.cnpq.br/0520491056597962; Mendonça, Elismauro Francisco de; https://orcid.org/0000-0002-6751-3279; http://lattes.cnpq.br/2305019128015847; Lima, Sheila Coelho Soares; https://orcid.org/0000-0002-6742-3708; http://lattes.cnpq.br/7690812849140276; Peterle, Gabriela Tonini; https://orcid.org/0000-0001-7735-936X; http://lattes.cnpq.br/6804646896381238; Silva, Adriana Madeira Alvares da; https://orcid.org/0000000280780304; http://lattes.cnpq.br/6445492335035108Oral squamous cell carcinoma (OSCC) has an estimated 377,713 new cases each year in the 2020-2022 triennium worldwide. Individuals affected by OSCC exhibit heterogeneous clinical behavior and diagnosis at an advanced stage, causing a worse prognosis. Early detection of the disease increases survival rates by up to 80%. Thus, the analysis of the dynamics between the immune response, through the analysis of the tumor lymphocytic infiltrate (TIL), and the tumor progression, evaluated by the use of the biomarkers PLK1 and FOXM1, can be essential tools to assist in the detection, recurrence and prognosis. of the disease. Serum PLK1 promoter region methylation, PLK1 and FOXM1 gene expression in tumor tissue, and Plk1 expression in tumor adjacent epithelium, dysplasia and tumor were evaluated as prognostic indicators in patients with OSCC. Plk1 expression was evaluated in 109 paraffinized tissue samples, 21 frozen tumor fragments and 30 serum (17 before treatment initiation and 13 after treatment). The Chi-Square and Fisher's Exact tests were used to establish an association between the variables studied with TIL density, methylation status and Plk1 expression; to compare the mean expression of Plk1 in the segments studied, the Mann-Whitney U test and the Bonferroni post-test were used; One-Way analysis of variance (ANOVA) and application of Dunnett and/or Tukey tests were applied to verify patterns of gene expression; and survival curves were evaluated using the Kaplan-Meier model. Low TIL density was associated with T3/T4 tumor size (p = 0.001) and clinical stage III/IV (p = 0.011); while high TIL was associated with T1/T2 tumor size (p = 0.001), clinical stage I/II (p = 0.01) and surgery as treatment performed (p = 0.046). The PLK1 methylation status showed no association with the variables analyzed. High expression of PLK1 was observed in 11 samples in relation to the control (p < 0.0001), as well as two samples had a different expression when in parity to the others (p < 0.0001). FOXM1 did not show expression in the sample group. The analysis of Plk1 expression in the segments showed a correlation between the adjacent epithelium pairs and the dysplasia and tumor regions (p < 0.001). High Plk1 expression was associated (p < 0.001) with the variables T3/4 tumor size, lymph node metastasis and stage III/IV. Likewise, low expression was associated (p < 0.001) with smaller tumor size, no lymph node involvement and early stages. Tumor Plk1 expression and TIL density were not shown to be prognostic factors when analyzed for the overall survival and disease free survival curves, but they did show to be related to tumor development and progression in OSCC.