Mestrado em Biotecnologia
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- ItemVariantes do gene NOTCH1 como biomarcadores para transtornos neurocognitivos(Universidade Federal do Espírito Santo, 2025-03-26) Rodrigues, Alda dos Santos; Paula, Flávia; https://orcid.org/0000-0001-8679-2982; http://lattes.cnpq.br/7913201450663683; Errera, Flávia Imbroisi Valle; https://orcid.org/0000-0002-8069-6372; http://lattes.cnpq.br/9337327437538048; https://orcid.org/0000-0002-8069-6372; http://lattes.cnpq.br/5759084443362110; Santos, Eldamaria de Vargas Wolfgramm dos ; https://orcid.org/0000-0003-3815-0760; http://lattes.cnpq.br/4688343262832362; Barcelos, Estevão Carlos Silva; https://orcid.org/0000-0001-7076-6165; http://lattes.cnpq.br/6899125943013073Neurocognitive disorder (NCD) can be mild (TNCL) when it does not impact independence in activities of daily living or major (TNCM), also known as dementia, is clinically heterogeneous and has been associated with different risk factors, such as metabolic diseases (diabetes, obesity), stroke, tobacco and alcohol use, among others. Genetic factors have been investigated, but are not yet fully known. Thus, the search for biomarkers that can be used in diagnosis, risk screening, prognosis and response to treatment is very important. In this context, the NOTCH1 gene is expressed in several tissues, mainly in the cerebral cortex, hippocampus and cerebellum, hematopoietic stem cells, heart and blood vessels, and is associated with 118 phenotypes. NOTCH1 encodes a protein homologous to the neurogenic NOTCH locus in humans (Homo sapiens), important for glucose homeostasis, lipid metabolism, cell growth and survival. The NOTCH signaling pathway is very important in neurogenesis and maintenance of adequate neuronal function in the human brain and has been implicated in NCD//dementia, including Alzheimer's disease (AD). Recent evidence points to a shared genetic architecture between metabolic alterations, NOTCH1 and NCD. NOTCH1 is expressed in the mature human hippocampus, and shows increased immunoreactivity in AD, and in cases with tau-positive Pick bodies. The absence of NOTCH1 is capable of affecting a cascade of secondary messengers associated with plasticity and spatial learning, and with reduced volume of brain regions, being associated with AD and, therefore, with the risk of NCD. Thus, the hypothesis tested in this work is that variants in this gene are associated with NCD. The objective is to verify whether variants in NOTCH1 are associated with NCD. Participants in the PAHO study "Health, Well-being and Aging" (SABE) from São Paulo, with complete data (N=1103), were classified for NCD using the score obtained in the Mini Mental State Examination (MMSE). Participants who scored below the MMSE cutoff (13 points) were considered cases [carriers of NCD (N=152)] and the remaining participants (951), with a score of 13 or higher, were used as controls. Clinical data were obtained from a standardized questionnaire and genetic data from whole-genome sequencing to identify genetic variants. Genotypes for the variants and their respective frequencies were obtained from the Brazilian Online Mutation Archive (ABraOM). Females comprised the majority of the population, corresponding to 64.09% (N=707). The prevalence of NCD was 13.78% (152 patients). The variants rs10870087, rs2990585, rs7874114 and rs7864342 associated with NCD were identified in DNA regulatory regions, mainly in active enhancers, suggesting their role in the regulation of gene expression, especially in brain tissues. The rs2990585 and rs7874114 variants have epigenetic marks (H3K27ac and H3K4me1), indicating involvement in the activation of genes related to synaptic plasticity and neurodevelopment. rs2990585 with a high probability of regulatory function (1f), binding to the CEBPA gene, involved in cell differentiation and inflammatory response. Enhancer 2 of this variant is active in several brain regions, such as the midbrain and frontal cortex. Furthermore, the rs10870087, rs7864342 and rs7874114 variants demonstrated eQTL activity for CYSRT1 in breast tissue, while rs2990585 presented eQTLs in genes such as NOTCH1 (blood and tibial artery) and NRARP (lymphocytes). These findings suggest that variants in NOTCH1 have an impact on gene regulation and are promising candidates for use as biomarkers of NCD. However, functional studies and studies in other populations are needed for generalization and application of these results
- ItemFluorescência da clorofila a como ferramenta indicadora de estresse mineral em Schinus terebinthifolia Raddi(Universidade Federal do Espírito Santo, 2025-03-31) Dalla, Rhaira Rodrigues; Guimarães, Marco Cesar Cunegundes; https://orcid.org/0000-0003-2146-0180; http://lattes.cnpq.br/0261991057482057; https://orcid.org/; Errera, Flavia Imbroisi Valle; https://orcid.org/0000-0002-8069-6372; http://lattes.cnpq.br/9337327437538048; Medeiros, Marcos Barros de; https://orcid.org/0000-0002-1633-3227; http://lattes.cnpq.br/6453244305834445The aim of this study was to evaluate the effectiveness of chlorophyll a fluorescence as a diagnostic tool to identify and monitor abiotic stress in Schinus terebinthifolia Raddi, which occurs due to nutrient deficiency or excess, affecting its growth and metabolism. The deficiency reduces photosynthesis and development, while excess can cause toxicity. This results in chlorosis, necrosis and reduced resistance to pathogens, compromising its adaptation and regeneration. In addition, we sought to correlate changes in fluorescence indices and the modulation of proteins involved in the protection and adaptation of the photosynthetic apparatus. In vivo analyses of chlorophyll a fluorescence were performed using a portable fluorometer at four stations located in Espírito Santo (AE1, AE2, AE3 and AE4) in plants under abiotic stress conditions. These analyses allowed obtaining parameters such as maximum photosynthetic efficiency (Fv/Fm) and non-photosynthetic energy dissipation (NPQ), evaluated by OJIIP and SPAD. In addition, a search was conducted at NCBI for potential proteins to be used in the development of diagnostic tests in plants under mineral stress. The results of the study demonstrated that the exposure of Schinus terebinthifolia Raddi to different abiotic stress conditions caused marked changes in photosynthetic efficiency and a significant reduction in maximum photosynthetic efficiency (Fv/Fm) indices, indicating damage to the photosynthetic system resulting from the applied stresses. The increase in energy dissipation via NPQ evidences functional changes in the photosynthetic system that occur early in the face of mineral stress conditions suffered by the red pepper plant. Furthermore, five proteins, OHP1, OHP2, PSBS, ELIP1 and cpSRP43, were identified as potential proteins to be tested in a greenhouse using Arabidopsis thaliana as a model organism. Thus, the study showed that chlorophyll a fluorescence is a promising tool for the diagnosis of abiotic stress in Schinus terebinthifolia Raddi, allowing the early detection of imbalances in the photosynthetic apparatus. Furthermore, the proteins mentioned can be applied in later studies because they are associated with the assembly of the photosynthetic apparatus and interaction with pigments for the synthesis of chlorophyll
- ItemO papel da L-arginina sobre o sistema oxidativo em células mda-mb-231 de câncer de mama triplo negativo(Universidade Federal do Espírito Santo, 2025-03-18) João Augusto Diniz Moura; Gouvea, Sonia Alves; https://orcid.org/0000-0001-5180-471X; http://lattes.cnpq.br/7268228122543743; Meira, Débora Dummer; Sartório, Carmem LuízaL-Arginine is a semi-essential amino acid whose role in cancer has been under investigation, showing beneficial results due to its involvement in various physiological systems. Therefore, it is necessary to investigate which pathways involving this amino acid directly contribute to tumor regression, such as those related to oxidative stress. In this context, the use of the MDA-MB-231 triple-negative breast cancer cell line, known for its high aggressiveness and limited treatment options due to the lack of therapeutic targets, highlighted the effects of different concentrations of L-ARG on cell viability and the oxidative system. The treatment involved exposure to L-ARG at concentrations of 800 µg/mL, 1600 µg/mL, and 3200 µg/mL for 48 hours. A dihydroethidium (DHE) staining assay was performed to detect the increased presence of superoxide anion (O₂⁻). The AlamarBlue assay was employed to measure cell proliferation and assess cell viability. AOPP values were determined according to the protocol described by Witko-Sarsat et al. (1996) to evaluate the production of advanced oxidation protein products. Western blotting was used to determine the influence of L-arginine treatment on SOD1 protein expression in MDA-MB-231 cells across the NT, 800 µg/mL, 1600 µg/mL, and 3200 µg/mL groups. Data were analyzed using GraphPad Prism® version 8.0.2 and corrected by Tukey’s test. The main findings of the study were: (I) Reduced cell viability; (II) Increased production of superoxide anion radicals (O₂⁻) at all concentrations; (III) Elevated levels of advanced oxidation protein products; and (IV) Reduced SOD1 expression. Therefore, through the significant results that contributed to elucidating the effects of L-arginine on the oxidative system in MDA-MB-231 cells, this study opens new perspectives with potential translational implications.
- ItemIdentificação de potenciais biomarcadores genéticos da rede de inflamação e seus riscos de desenvolver dispneia pós-COVID-19(Universidade Federal do Espírito Santo, 2024-02-24) Ventorim, Vinicius do Prado; Meira, Débora Dummer; https://orcid.org/0000-0002-6092-2459; http://lattes.cnpq.br/7199119599752978; Louro, Iúri Drumond ; https://orcid.org/0000-0001-5160-9615; http://lattes.cnpq.br/3817361438227180; https://orcid.org/0009-0001-9958-5023; http://lattes.cnpq.br/2427358142499457; Paula, Flávia; https://orcid.org/0000-0001-8679-2982; http://lattes.cnpq.br/7913201450663683; Carvalho, Elizeu Fagundes; https://orcid.org/0000-0003-4620-7253; http://lattes.cnpq.br/2742420738858309The study aimed to identify genetic biomarkers associated with the progression of dyspnea in individuals post-COVID-19. Following approval by the Research Ethics Committee (CEP), a case-control study was conducted involving 277 individuals. Participants were selected based on clinical and epidemiological criteria, allowing a representative sample of the study population. For genetic analysis, whole-exome sequencing was performed, the identification of genetic variants potentially associated with the clinical outcome. Rigorous statistical methodologies were applied to select the most relevant variants, focusing on determining the allelic risk for the progression of dyspnea. The prevalence of dyspnea among participants was 15.16%, underscoring the importance of investigating genetic factors that may influence its manifestation. Eleven genetic variants with p-values lower than 0.05 were identified, suggesting a statistically significant association with the risk of dyspnea progression. The identified variants are located in ten genes that play key roles in the immune system, in the regulation of vascular hemostasis, and in the response to epithelial damage. These findings indicate that genetic predisposition may contribute to the persistence of dyspnea in individuals who have overcome the acute phase of COVID 19. Gene analysis revealed a strong correlation with critical biological processes in the inflammatory response and tissue recovery. The immune system plays a central role in regulating the persistent inflammation observed in some post-COVID-19 individuals, and may influence the maintenance of chronic cardiorespiratory symptoms. Vascular hemostasis, on the other hand, may be associated with coagulation disorders previously described in patients with COVID-19, impacting pulmonary perfusion and contributing to dyspnea. Genes related to epithelial damage are possibly involved in lung tissue repair, being decisive for the functional recovery of the organ after infection. These results provide valuable directions for future clinical and epidemiological research in the State of Espírito Santo (Brazil). The identification of genetic biomarkers can help in the risk stratification of patients, allowing for more personalized and targeted therapeutic approaches. Moreover, the findings reinforce the need for further investigations to understand the mechanisms by which these genetic variants influence the evolution of post-COVID-19 dyspnea. Furthermore, this study contributes to the growing body of knowledge regarding sequelae of COVID-19 and its genetic bases, highlighting the importance of the integration between genomics and epidemiology for the development of public health strategies. Based on these findings, it is expected that further research can validate the identified biomarkers and deepen the understanding of the genetic factors involved in the recovery of post COVID-19 patients. This approach may, ultimately, lead to the development of more effective interventions to mitigate the impacts of persistent dyspnea on the quality of life of affected individuals
- 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