Doutorado em Biotecnologia

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Navegando Doutorado em Biotecnologia por Assunto "Biomarcador"

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    Novas abordagens no diagnóstico não invasivo de câncer bucal utilizando espectroscopia ftir em amostras de sangue e saliva
    (Universidade Federal do Espírito Santo, 2021-09-29) Thebit, Marcela Marçal; Gouvea, Sonia Alves; https://orcid.org/000000015180471X; http://lattes.cnpq.br/7268228122543743; https://orcid.org/0000-0002-7305-5753; http://lattes.cnpq.br/7117284804597883; Abreu, Gláucia Rodrigues de; https://orcid.org/0009-0008-8772-8470; http://lattes.cnpq.br/0229590907405570; Endlich, Patrick Wander; https://orcid.org/0000-0002-7597-3603; http://lattes.cnpq.br/9669721267985878; Carvalho, Luis Felipe das Chagas e Silva de; https://orcid.org/0000-0003-1063-4624; http://lattes.cnpq.br/5669726117269568; Silva, Adriana Madeira Alvares da; https://orcid.org/0000000280780304; http://lattes.cnpq.br/6445492335035108
    Oral cancer is usually diagnosed through invasive methods, for although systemic changes can be easily detected, the identification of an effective systemic biomarker using a simple and affordable approach has yet to take place. Attenuated total reflection Fourier transform infrared (ATR-FTIR) appears as an alternative for the chemical and structural characterization of organic and inorganic materials, including liquid samples as serum, plasma and saliva and a possible bridge between molecular analysis and clinical practice. Objective: The aim of this study was to evaluate if ATR-FTIR spectroscopy can be applied as a tool in oral cancer screening through the analysis of plasma and salivary samples. Methodology: Plasma and saliva samples from oral cancer patients and controls were analyzed by ATR-FTIR spectroscopy. Experimental data were fed into the MATLAB 2018b software and them analyzed through two different routines in order to identify points of differentiations between the groups. Results: Biochemical analysis of the plasma samples through ATR-FTIR yielded 92% sensitivity and 100% specificity in differentiating between groups. The biggest difference was found between peaks that might be related to nucleic acids and nucleic acid phosphate. Pilot study with salivary samples reached similar accuracy Conclusion: We conclude that ATR-FTIR successfully discriminated between oral cancer patients and healthy subjects using plasma samples, with nucleic acids being found as major biomarkers.
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    Potencial prognóstico de células inflamatórias e PD-L1 solúvel em carcinoma epidermoide de cabeça e pescoço
    (Universidade Federal do Espírito Santo, 2024-04-16) Daniel, Camila Batista; Co-orientador1; https://orcid.org/; http://lattes.cnpq.br/; Co-orientador2; https://orcid.org/; http://lattes.cnpq.br/; Co-orientador3; https://orcid.org/; http://lattes.cnpq.br/; Co-orientador4; ID do co-orientador4; Lattes do co-orientador4; Zeidler, Sandra Lúcia Ventorin von ; https://orcid.org/0000-0002-8897-5747; http://lattes.cnpq.br/5785612863130498; Orientador2; https://orcid.org/; http://lattes.cnpq.br/; https://orcid.org/; http://lattes.cnpq.br/; Santos, Eldamária de Vargas Wolfgramm dos ; https://orcid.org/; http://lattes.cnpq.br/; Errera, Flavia Imbroisi Valle ; https://orcid.org/; http://lattes.cnpq.br/; Arantes, Lidia Maria Rebolho Batista ; https://orcid.org/; http://lattes.cnpq.br/; Mendonça, Elismauro Francisco de ; http://lattes.cnpq.br/; 5º membro da banca; https://orcid.org/; http://lattes.cnpq.br/; 6º membro da banca; https://orcid.org/; http://lattes.cnpq.br/; 7º membro da banca; https://orcid.org/; http://lattes.cnpq.br/
    Immunosuppression is recognized as a hallmark of cancer and has been associated with worse outcomes. In head and neck squamous cell carcinoma (HNSCC), immunosuppression is a hallmark of the tumour and may be mediated by immunosuppressive inflammatory cells and PD-L1 expression. Given their participation in this process, our study aimed to describe the prognostic impact of these elements in tumour tissue and peripheral blood HNSCC patients. We conducted a multicentre retrospective study with biological samples and clinical data from HNSCC patients recruited at two centres in Brazil and the United Kingdom and healthy individuals. To analyse the inflammatory infiltrate, we used tumour tissue slides stained with haematoxylin and eosin and immunohistochemistry for PD-L1 analysis and neutrophil quantification. Absolute leukocyte counts were retrieved from pretreatment blood counts available in medical records. To evaluate the prognostic value of PD-L1 in liquid biopsy, we used serum and plasma samples obtained from patients and healthy individuals for quantification of soluble PD-L1 (sPD-L1) by ELISA. Fresh tumour tissue samples were used to analyse CD274 gene expression levels using the RT-qPCR technique. Furthermore, we analysed PD-L1 expression by flow cytometry in HNSCC cell lines and quantified sPD-L1 levels in the supernatant. The tumour microenvironment analysis showed that low levels of lymphocytes are associated with worse overall survival and disease-free survival. However, we did not observe this relationship with tumour-associated neutrophils and tumour PD-L1. When analysed together in a scoring system, we demonstrated that low levels of lymphocytes, high expression of PD-L1 and high infiltration of neutrophils are associated with a worse outcome. In blood, a high ratio of neutrophils and lymphocytes (NLR) was also associated with worse survival but was not correlated with inflammatory components of the tumour. High levels of sPD-L1 protein were associated with reduced overall survival, however our study did not identify a relationship between sPD-L1 levels in the blood and PD-L1 expression in tumour tissue, determined by immunohistochemistry and RT-qPCR. In contrast, the in vitro study showed that the levels of sPD-L1 released 14 into the supernatant are strongly correlated with cytoplasmic and membrane expression. Our data further showed that sPD-L1 levels in patients were positively correlated with the number of leukocytes, neutrophils and monocytes in peripheral blood. Taken together, our results highlight the prognostic potential of markers of the tumour microenvironment analysed in a combined manner, in a scoring system, as a way of providing a more comprehensive overview of tumour behaviour, highlighting important events, such as immunosuppression. Our results highlight the prognostic potential of PD-L1 detected by liquid biopsy in HNSCC and indicate that the levels detected in the blood do not necessarily correspond to what is observed in the tumour. Therefore, it is believed that it should be analysed as an independent marker whose immunosuppressive role is carried out at a systemic level