Doutorado em Biotecnologia

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Navegando Doutorado em Biotecnologia por Autor "7º membro da banca"

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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
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    Transmissão por insetos do complexo papaya meleira virus (PMeV e PMeV2) e prospecção e caracterização de uma fração da proteína estrutural do PMeV
    (Universidade Federal do Espírito Santo, 2024-05-06) Almeida, Joellington Marinho de; 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; Orientador1; https://orcid.org/; http://lattes.cnpq.br/; Orientador2; https://orcid.org/; http://lattes.cnpq.br/; https://orcid.org/; http://lattes.cnpq.br/; 1º membro da banca; https://orcid.org/; http://lattes.cnpq.br/; 2º membro da banca; https://orcid.org/; http://lattes.cnpq.br/; 3º membro da banca; https://orcid.org/; http://lattes.cnpq.br/; 4º membro da banca; 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/
    Papaya blight is a disease caused by a virus that can lead to a loss of productivity in Carica papaya orchards in producing countries such as Brazil, Mexico, Ecuador and Australia where the disease occurs. Infection of the plant causes burning of the tips of young leaves, spontaneous exudation of latex and spots on the fruit. The latex oxidizes in the presence of air, giving rise to a "honey-like" appearance, which together with other factors interferes with the commercial acceptance of the fruit. However, the vector that transmits the papaya late blight disease is still unknown in all producing countries. Studies carried out by various research groups have pointed to the leafhopper as a potential disseminator of the disease, in addition to the relationship with fungi, seeds and mechanical transmission. In Brazil, it is caused by the viral complex papaya meleira virus (PMeV) and papaya meleira virus 2 (PMeV2), and the coexistence of the two viruses has proven that PMeV2 is encapsidated by PMeV, and it is known that PMeV has two ORFs responsible for encoding structural proteins that make up the capsid (ORF1) and a putative RdRp protein (ORF2). Knowing that the structural proteins of PMeV are responsible for encapsidating the two genetic materials, the use of the expression of a recombinant protein fraction of the capsid protein (CP) of PMeV could become viable in the development of rapid tests, identification of viral particles in different locations of the plant and potential application in studies of virus-plant-vector transmission. In this study, we developed material on understanding the viral transmission of the papaya meleira virus complex by insects using the available literature and the prospecting and characterization of a fraction (p441) of the PMeV structural protein. The p441 fraction of the PMeV capsid protein was expressed in E. coli BL21(DE3) and extracted from the SDS-PAGE gel. The protein fraction was expressed at four different times (30 minutes, 1 hour, 2 hours and 4 hours) to check which would correspond to its highest production and its extracted fraction was used to produce polyclonal antibodies. Based on the available sequence, an in silico characterization was carried out to determine ab initio secondary and tertiary structure models, physicochemical parameters and the prediction of immunogenic peptides that could be identified in its primary sequence. The results obtained open up possibilities for the design of tests that are effective in detecting PMeV, based on observations made about the protein structure of the p441 fraction as being a stable protein with low thermal mobility. In addition, 18 peptides found on its surface have been shown to be capable of triggering an immune response and to be a region that can be used to produce antibodies that can be applied in future studies to locate the virus in different parts of the plant. Finally, it was found that the conditions of expression in a time of 4 hours obtained the best response, generating protein concentrations of around 633 to 923 mg/mL verified after SDS-PAGE gel extraction