Mestrado em Biotecnologia
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- ItemAnálise de variantes nos genes ACE1 e LRP1 e sua associação com a doença de Alzheimer na Grande Vitória/ES : potenciais biomarcadores genéticos para diagnóstico complementar(Universidade Federal do Espírito Santo, 2025-02-26) Lopes, Victor Alves; Paula, Flávia de ; https://orcid.org/0000-0001-8679-2982; http://lattes.cnpq.br/7913201450663683; https://orcid.org/0009-0006-1332-6079; http://lattes.cnpq.br/1133525392804189; Errera, Flavia Imbroisi Valle; https://orcid.org/0000-0002-8069-6372; http://lattes.cnpq.br/9337327437538048; Maranduba, Carlos Magno da Costa ; https://orcid.org/0000-0001-7327-1934; http://lattes.cnpq.br/4763153859701731The accelerated aging of the Brazilian population has contributed to the increasing prevalence of chronic diseases such as Alzheimer’s disease (AD), a multifactorial condition involving complex interactions between genetic and environmental factors. This study aimed to analyze genetic variants in the ACE1 (INDEL rs4646994) and LRP1 genes and their association with AD in a population from Greater Vitória, Espírito Santo, to identify potential genetic biomarkers for complementary diagnosis. A cross-sectional, analytical, and observational study was conducted with 246 participants clinically diagnosed with sporadic AD, matched by sex, age, and ethnicity. Blood samples were collected for genomic DNA extraction, followed by PCR and agarose gel electrophoresis for genotyping. The results showed no significant association between the INDEL variant of the ACE1 gene and AD or the APOE ε4 allele status, suggesting a limited contribution of this variant in the studied population. Conversely, the LRP1 gene showed a significant association, with the D allele (DD+ID) displaying a potentially protective role against AD. These findings reinforce the multifactorial nature of AD and highlight the influence of regional genetic characteristics, considering the historical diversity and genetic admixture of the Espírito Santo population. This study contributes to understanding genetic differences between populations and supports the development of personalized medicine strategies for the diagnosis and treatment of AD
- ItemBioprospecção do fucoidan e ouabaína: efeitos na motilidade e vitalidade espermática humana in vitro(Universidade Federal do Espírito Santo, 2025-03-18) Silva, Paula Gabriella Pedras; Nogueira, Breno Valentim; Meira, Débora Dummer; https://orcid.org/0000-0002-6092-2459; Paula, Flavia de; Maranduba, Carlos Magno da CostaOral hormonal contraceptives (OC) are widely used in family planning but pose significant health risks to women. Although effective, they can cause severe adverse effects, such as cardiovascular alterations that increase the risk of pulmonary embolism, venous and arterial thrombosis, and metabolic diseases such as diabetes and dyslipidemia. Cardiovascular diseases (CVD) are the leading cause of death in Brazil and globally, and hormonal contraceptive methods are associated with an increased risk of cerebrovascular accidents and other CVDs. Plant-based spermicides are alternatives to chemical spermicides like Nonoxynol-9 (N-9), which can cause vaginal mucosa damage and increase the risk of infections. Alternatives to N-9, such as Sodium Lauryl Sulfate (SLS), also present adverse effects, including skin and mucosal irritation. Fucoidan, a sulfated polysaccharide found in brown seaweeds, has beneficial biological activities and is investigated as a drug carrier due to its low toxicity. Ouabain, extracted from the plant Strophanthus gratus, acts on sperm capacitation and inhibits Na+/K±ATPase, playing a significant role in various physiological and pathological processes. This study aimed to analyze the spermicidal activity of fucoidan, extracted from the seaweed Fucus vesiculosus, and ouabain on human sperm in vitro. The methodology involved collecting semen samples from volunteers aged 18 to 45 years, after approval from the Ethics and Research Committee under the Ethical Appreciation Presentation Certificate (CAAE) 79783124.5.0000.5060, which were subjected to treatments with negative control, positive control (SLS), fucoidan, ouabain, and a combination of both. Sperm motility and vitality were assessed at two different time points (0 and 15 minutes) using seminal analysis techniques standardized by the World Health Organization. The treatments significantly reduced sperm motility and vitality compared to the positive control (SLS). The combination of fucoidan and ouabain showed the greatest reduction in sperm motility, with approximately a 28.3% decrease in progressive motility and a 28.1% decrease in vitality, suggesting a synergistic effect between the compounds. Statistical analysis using non-parametric tests of Kruskal-Wallis and Wilcoxon post hoc confirmed the significance of these reductions at times T0 and T15. Moreover, the interaction of the compounds with reactive oxygen species (ROS) suggests a mechanism of action that compromises cellular integrity and sperm function. These findings highlight the potential of fucoidan and ouabain as effective spermicidal agents, contributing to the development of safer and more efficient contraceptive methods. However, it is necessary to consider the costs associated with each treatment and conduct further research to confirm these hypotheses and explore their future biotechnological applications
- ItemClassificação de gravidade e identificação de biomarcadores na Covid-19: análise do exoma de pacientes através de máquinas de vetores de suporte com kernel linear (SVM)(Universidade Federal do Espírito Santo, 2025-02-24) Zetum, Aléxia Stefani Siqueira; Meira, Débora Dummer; https://orcid.org/0000-0002-6092-2459; http://lattes.cnpq.br/7199119599752978; Louro, Iuri Drumond; https://orcid.org/0000-0001-5160-9615; http://lattes.cnpq.br/3817361438227180; https://orcid.org/0000-0002-5086-411X; Paula, Flávia de; https://orcid.org/0000-0001-8679-2982; Carvalho, Elizeu Fagundes de; https://orcid.org/0000-0003-4620-7253Introduction: SARS-CoV-2 infection presents a wide spectrum of clinical manifestations. Genetic variations may influence the host's response to the virus. The use of Machine Learning (ML) has shown promise in identifying genetic biomarkers and individuals who may develop severe forms of the disease. Objective: To develop an ML model using exome data to predict clinical outcomes in COVID-19 patients and identify genes potentially associated with disease severity. Methodology: The study involved data from 239 COVID-19 patients ("Non-severe" and "Severe"). DNA sequencing was performed, and ancestry analysis was conducted. A Support Vector Machine (SVM) model with a linear kernel was developed to predict COVID-19 severity, utilizing Recursive Feature Elimination (RFE) to select the most influential variants. Metrics such as Area Under the Curve-Receiver Operating Characteristic (AUC-ROC), accuracy, F1 score, sensitivity, and specificity were used. Subsequently, logistic regression (LR) analysis was performed with the variants selected by SVM-RFE and confounding variables. Results and Discussion: The SVM model with a linear kernel achieved an AUC-ROC of 0,81, accuracy of 83%, and an F1 score of 0,78, indicating a good capacity to discriminate between "Severe" and "Non-severe" cases of COVID-19. Fifteen variants were selected by the model, of which seven were significantly associated with disease severity in the LR analysis. Risk variants include WSCD1 (rs2302837 "A/A" or "A/G," 95% CI: 1,32–7,24, OR: 3,09, P < 0,01), PTPRS (rs1143700 "A/A" or "A/G," 95% CI: 1,54–7,07, OR: 3,30, P < 0,01), ARVCF (rs2073744 "A/A" or "A/G," 95% CI: 1,31–6,30, OR: 2,88, P < 0,01), and LVRN (rs10078759 "G/G" or "G/C," 95% CI: 1,07–4,31, OR: 2,08, P = 0,04). Conversely, protective variants include ALDH4A1 (rs6426813 "G/G" or "G/A," 95% CI: 0,23–0,93, OR: 0,48, P = 0,02), ARHGAP22 (rs10776601 "C/C" or "C/T," 95% CI: 0,09–0,56, OR: 0,23, P < 0,01), and C3 (rs423490 "A/A" or "A/G," 95% CI: 0,14–0,70, OR: 0,32, P < 0,01). The results demonstrated that the SVM with a linear kernel is effective in predicting COVID-19 severity using exome data. The protein-protein interaction (PPI) network analysis identified biological pathways associated with the immune system, inflammatory response, and blood coagulation. Genes such as C3, PTPRS, and LVRN stood out in functions related to immune response regulation and inflammation modulation, suggesting these pathways are directly linked to adverse COVID-19 outcomes. The network also revealed the interconnection between cellular signaling processes and stress response mechanisms, which may explain the variability in clinical responses observed among patients. Conclusion: The SVM with a linear kernel using our data proved effective in predicting COVID-19 severity. This study highlights the importance of integrative approaches to better understanding the disease. Identifying genetic biomarkers can aid in treatment and management of future pandemics
- ItemEfeitos do tratamento com L-Arginina sobre a inflamação em células MDA-MB-231 de câncer de mama triplo negativo(Universidade Federal do Espírito Santo, 2025-03-21) Maurício, Lorena Souza Rittberg; Gouvea, Sônia Alves; https://orcid.org/0000-0001-5180-471X; http://lattes.cnpq.br/7268228122543743; https://orcid.org/0009-0000-9398-1423; http://lattes.cnpq.br/9481300734961481; Errera, Flávia Imbroisi Valle; https://orcid.org/0000-0002-8069-6372; http://lattes.cnpq.br/9337327437538048; Amorim, Girlandia Alexandre Brasil; https://orcid.org/0000-0002-5455-7141; http://lattes.cnpq.br/1402295792093274Triple-negative breast cancer (TNBC) is an aggressive form of breast cancer lacking hormone receptors or HER2, which limits therapeutic options and worsens prognosis. Given the scarcity of therapeutic targets, the search for strategies that influence the cellular mechanisms involved in tumor progression has been investigated. In this context, L-arginine, a conditionally essential amino acid, has stood out for its potential to regulate inflammation and tumor metabolism, which may, in turn, positively influence the response to treatment. The main objective is to evaluate the effects of L-arginine on cell viability and inflammation-related pathways in MDA-MB 231 cells. MDA-MB-231 cells were subjected to different concentrations of L-arginine (800ug/ml; 1600ug/ml; and 3200ug/ml) for 48h. The AlamarBlue® assay was used to measure proliferation and evaluate cell viability. To evaluate the expression of Nrf2 and NF-kB, the Western Blot method was performed and inflammatory cytokines were quantified by flow cytometry. The results showed that L-arginine decreased the cell viability of MDA MB 231 cells at higher concentrations of 1600ug/ml; and 3200ug/ml there was an increase in the expression of Nrf2 and NF-kb, which can activate the ferroptosis pathway; increased levels of cytokines such as IL-17A, IL-6, IL-2 and IFN-γ; decreased IL-10 and there were no significant changes between the IL-4 and TNF-α groups. These findings suggest that L-arginine has potential as a therapeutic strategy in breast cancer, especially in the context of aggressive tumors such as TNBC
- ItemAvaliação do efeito de citotoxicidade dos microplásticos na linhagem mamária humana MCF10A(Universidade Federal do Espírito Santo, 2025-03-10) Silva, Laíza dos Santos Ribeiro da; Co-orientador1; Santos, Eldamária de Vargas Wolfgramm dos; https://orcid.org/0000-0003-3815-0760; Paula, Flávia de; Tavares, Marcella PortoIntroduction: It is known that the use of plastic and its derivatives has been growing exponentially, accompanied by advances involving poor management, detachment and degradation of plastic waste. Studies also indicate the presence of components such as microplastic in the human body, which may pose health risks. Furthermore, compounds like resveratrol (RSV) have been extensively studied for their antiinflammatory and anticancer properties, as well as their interactions with various substances. Given this, investigating the impacts of microplastics and compounds such as resveratrol is essential to understanding the implications for human health. Objective: To evaluate the impact of microplastics on MCF10A cells and the influence of resveratrol in this context. Methodology: This is a laboratory study using MCF10A cell culture exposed to six different concentrations of MPs (20 μm and 1 μm), RSV exposure at six different concentrations, and a mixed group (combining 20 μm MPs and RSV) at different concentrations. Cell viability was analyzed at 24h and 72h time points through an MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide). Results: The isolated exposure of cells to MPs (20 and 1 μm) did not show significant changes in cell viability at any time point. However, an increase in cell proliferation was observed when exposed to 1 μm MPs. Regarding RSV, cell viability was reduced to 11.74% after 72h of exposure at a concentration of 500 μM. In the mixed treatment, cell viability was reduced to 46.90% (p<0.0025) with 500 μg/mL MPs + 250 μM RSV and to 11.36% (p<0.0001) with 1000 μg/mL MPs + 500 μM RSV, suggesting a potential interaction between the compounds. Furthermore, the IC50 of RSV was identified as 263.8 μM at 72h. Conclusion: The results showed that the microplastics (20 and 1 μm) did not cause significant cytotoxic effects, whereas resveratrol (RSV), at high concentrations, reduced cell viability, with an IC50 of 263.8 μM. The combination of microplastics and RSV indicates a possible synergistic interaction, increasing cytotoxicity at higher concentrations. Given the limitations of the study, it is necessary to investigate whether chronic exposure to microplastics could increase the risk of diseases, including breast cancer, requiring new experimental conditions. Therefore, further studies are needed to better understand the impact of microplastics on human health and the mechanisms involved