Mestrado em Biotecnologia
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- ItemEfeito modulador da laser terapia sistêmica na inflamação em modelo de acidente vascular encefálico(Universidade Federal do Espírito Santo, 2024-10-18) Araújo, Stephanie Oliveira de; Pedrosa, Diego França ; https://orcid.org/; http://lattes.cnpq.br/; Nogueira, Breno Valentim ; https://orcid.org/0000-0002-7169-5634; http://lattes.cnpq.br/; https://orcid.org/; http://lattes.cnpq.br/4937376875703210; Bastos Filho, Teodiano Freire ; https://orcid.org/0000-0002-1185-2773; http://lattes.cnpq.br/; Silva, André Romero da ; https://orcid.org/0000-0002-1497-6093 ; http://lattes.cnpq.br/This master's dissertation analyzed the impacts of systemic photobiomodulation, through the non-invasive technique of Intravascular Laser Irradiation of Blood (ILIB), in an experimental model of global cerebral ischemia in mice. The main focus was to examine how this therapy acts in a dose-dependent manner in the recovery of brain injuries caused by bicarotid occlusion. The mice were exposed to this occlusion, after which ILIB was applied at a dose of 3.2 J (TL group) and 6 J (TH group), and the therapeutic effects were measured through functional and biochemical parameters. Motor evaluations of the mice were performed at two different times: before and after the induction of stroke, showing a significant improvement in the groups treated with ILIB, especially at the higher doses, improving the motor ability, strength and mobility of the mice, as well as a decrease in inactivity time. The macroscopic area of cerebral infarction was analyzed, and the treated group showed a reduction (p<0.01) in the area damaged by ischemia. In addition, blood samples were collected for analysis of blood cell counts, revealing a modulation in the inflammatory response in the groups that received ILIB (p<0.05). Plasma and brain were also extracted to measure oxidative stress (OS) levels and inflammatory markers, and in the TL and TH groups there was a modulation in the production of interleukins (IL-12p70, IL-6, IL-10, MCP-1 and IFN-γ); among these, there was a significant increase in anti-inflammatory interleukin and others that are responsible for the response to post-stroke infections; these effects were dose-dependent. A decrease in OS was observed in the TH group compared to the AVC group in the analysis of protein oxidation in plasma of 37% and in the brain of 35%, however, in lipid peroxidation it was 70% in plasma and 37% in the brain. It is noteworthy that ILIB in this model showed surprising results. Clinical studies should be carried out to standardize the dose in humans and the number of applications, which prove the effect, as well as clarify the specific mechanisms that allow ILIB to exert its neuroprotective effects
- ItemAnálise dos efeitos da ativação cerebral por eletroencefalografia em pacientes pós-AVC durante imagética motora, estimulação elétrica funcional e luva robótica(Universidade Federal do Espírito Santo, 2024-08-15) Souza, Fernanda Vaz de; Bastos Filho, Teodiano Freire ; https://orcid.org/0000-0002-1185-2773; http://lattes.cnpq.br/3761585497791105; https://orcid.org/0009-0005-0201-9260; http://lattes.cnpq.br/6822429982883146; Rodriguez, Denis Delisle ; https://orcid.org/0000-0002-8937-031X; http://lattes.cnpq.br/7140331839822423; Fiorin, Fernando da Silva; https://orcid.org/0000-0002-6517-7160; http://lattes.cnpq.br/1783079128689681Objective: to analyze the effects of brain activation by electroencephalography (EEG) in post-stroke patients during stimulation with Motor Imagery (MI), Functional Electrical Stimulation (FES) and Robotic Glove (RG). Methodology: EEG recordings of three (03) post-stroke subjects, with hand movement impairment during MI, FES, RG and combination of FES+RG, were analyzed and compared. Results: The analysis of the Mu rhythm demonstrated that therapies with RG, FES and FES+RG were able to significantly reduce the Relative Power (RP) of this rhythm in volunteer 1 (V1), when compared to IM therapy. In volunteer 2 (V2), we observed the same pattern, however, in volunteer 3 (V3) no statistical significance was observed in the application of FES and FES+RG, when compared to IM. Contrary to expectations, RG therapy in V3 demonstrated a pattern of elevation in the relative power of the Mu rhythm. However, when comparing RG with other therapies in this participant, significant and encouraging results were obtained. After analyzing the Beta Low rhythm, it was demonstrated that FES+RG therapy was superior and significant when compared to IM in the three individuals studied. FES-only therapy was also found to be significant in V2 and V3 compared to IM. During the Beta High frequency analysis, it was observed that FES+RG therapy was superior and significant when compared to IM in V1 and V3. V2 did not demonstrate statistically significant results. Conclusions: Relevant results were obtained using associated FES+RG stimulations. Therefore, it is concluded that the association of these therapies can assist in the post-stroke neuro-motor rehabilitation process, enabling the development of new neuro-rehabilitation protocols that are accessible and safe
- ItemNicotiana benthamiana como organismo modelo para estudos com o complexo viral causador da meleira do mamoeiro(Universidade Federal do Espírito Santo, 2024-03-25) Cunha, Esdras Mendes da; Fernandes, Patrícia Machado Bueno; https://orcid.org/; http://lattes.cnpq.br/; Fernandes, Antônio Alberto Ribeiro; https://orcid.org/0000-0002-8261-9717; http://lattes.cnpq.br/; https://orcid.org/; http://lattes.cnpq.br/; Xavier, André da Silva; https://orcid.org/; http://lattes.cnpq.br/; Araújo, Marlonni Maurastoni de; https://orcid.org/; http://lattes.cnpq.br/The papaya tree (Carica Papaya L.) is a plant native to the Americas and due to the economic value of papaya, the plant is produced in tropical areas around the world. One of the viruses that plague producers is papaya stick disease, which is caused by a viral complex, Papaya meleira virus and Papaya meleira virus 2. This work aims to establish the Nicotiana benthamiana plant as a model organism for future studies with the PMeV and PMeV2 viruses, the viral complex that causes papaya honey. N. benthamiana is the most used experimental host in plant virology, mainly due to the large number of diverse plant viruses that can infect it. Since, working with C. papaya presents significant challenges due to its biological complexity, especially related to its pre- and post-flowering time, with a flowering period that can take four to seven months. N. benthamiana in fact accumulates viral loads for sixty days, as occurs with longer-established viruses. During the entire period of sixty days after inoculation, infection levels are considerable to establish N. benthamiana as a host. The inoculum is passed between different individuals, maintaining the infectious capacity of the complex without the need to pass through a vector. The presence of viruses was not observed in plants derived from seeds of infected plants. The results obtained in this work indicate that N. benthamiana is a model organism for future studies of gene silencing in order to establish ways to control papaya honey.
- ItemModulação da atividade enzimática e estabilidade da tripsina porcina por carboidratos(Universidade Federal do Espírito Santo, 2024-06-27) Couto, Aurelio dos Santos; 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/Trypsin is a serine protease with catalytic activity sensitive to its environment, which limits its applications in complex biological systems, high temperatures, or extreme pH conditions. Modulating enzyme activity and stability with carbohydrates emerges as a promising strategy to overcome these restrictions. Thus, the modulation of porcine trypsin activity and stability by different carbohydrates (arabinose, L-rhamnose, beta cyclodextrin, maltose, and fructose) at varying concentrations (1:10, 1:30, 1:60, and 1:100 w/w) was investigated. Enzymatic activity was assessed through biochemical assays, while biophysical methods (UV spectroscopy, foldrate calculation, dynamic light scattering (DLS), and zeta potential) were employed to investigate conformational stability and enzyme supramolecular states. Arabinose and L-rhamnose significantly increased trypsin enzymatic activity at certain concentrations between 1:10 and 1:30 (w/w), whereas the other carbohydrates not only failed to increase activity but also decreased enzyme activity. UV spectroscopy did not reveal significant conformational differences in the protein after carbohydrate addition. Foldrate calculation indicated that trypsin samples with 1:10 w/w arabinose and 1:30 w/w L-rhamnose showed significant conformational changes, suggesting greater stability at these tested concentrations. Aggregate profiling showed an increase in particle diameter with 1:30 w/w L-rhamnose, suggesting interaction between the protein and carbohydrate. Regarding protein surface charge (zeta potential), 1:10 w/w arabinose and 1:30 w/w L-rhamnose exhibited higher readings but did not indicate the ability to prevent supramolecular state formation. L-rhamnose at 1:30 w/w is suggested as the optimal condition to optimize porcine trypsin activity without significantly affecting its colloidal stability. These findings may have important implications for trypsin use across various research and industrial applications. Future studies could explore the underlying molecular mechanisms of carbohydrate-induced trypsin modulation, elucidating the basis for developing more efficient enzyme stabilizers
- ItemPreservação de amostras de folhas do mamoeiro para detecção dos vírus da meleira (PMeV E PMeV2) por diagnóstico molecular(Universidade Federal do Espírito Santo, 2024-03-27) Santana, Ana Paula de Jesus; Ventura, José Aires ; https://orcid.org/0000-0003-1422-1739 ; http://lattes.cnpq.br/8687116881326074 ; https://orcid.org/; http://lattes.cnpq.br/5112084593175979 ; Silva, Diolina Moura ; https://orcid.org/0000-0003-3885-280X ; http://lattes.cnpq.br/0341541450627705 ; Berilli, Ana Paula Candido Gabriel ; https://orcid.org/0000-0002-9229-4653 ; http://lattes.cnpq.br/8154953381311097The papaya blight disease is caused by the papaya blight virus complex (PMeV and PMeV2). The strategy for controlling the disease is to identify symptomatic plants that are removed from the field using the roguing technique. However, there is often a need for molecular confirmation of plants suspected of being infected. Improving techniques for maintaining the virus in biological material, such as herborization, can help in time scale studies of viral genomics. The aim of this study was to evaluate a methodology for collecting, preserving and storing samples of papaya leaves infected by the blight virus complex for molecular diagnosis. The efficiency of herborization and storage in preserving papaya leaf samples for diagnosis was determined. The samples were collected manually by cutting young leaves, preferably with apical lesions characteristic of the symptoms of blight. A total of 48 samples were separated into two differently stored groups, 24 kept in an ultrafreezer and 24 herbarized, which in turn were subdivided into two subgroups with 12 asymptomatic samples (UM and HM) and 12 symptomatic samples (UA and HA). Analyses for the molecular diagnosis of the PMeV and PMeV2 viruses were carried out at intervals of zero (T0), five (T5), ten (T10), twenty (T20) and forty (T40) days after storage. It was observed that at T20 and T40 all the samples showed the presence of the PMeV viral complex in both forms of preservation. However, the analyses at times T5 and T10 did not obtain a uniform pattern for identifying the virus, possibly due to the non-uniform distribution of the viruses in the cells of the papaya leaves. The results validated an alternative methodology for recommending a new low-cost and easy-to-use sample preservation and storage protocol for the molecular diagnosis of papaya blight