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- ItemOvarian cancer chemoresistance: the role of inflammasomes in cisplatin resistance and the impact of mitochondrial reprogramming in parp inhibitor resistance(Universidade Federal do Espírito Santo, 2025-12-17) Souza, Josiany Carlos de; Rangel, Leticia Batista Azevedo; https://orcid.org/0000-0002-6150-4064; http://lattes.cnpq.br/6706127386696883; https://orcid.org/0000-0001-6777-0227; http://lattes.cnpq.br/1856469122580933; Silva, Adriana Madeira Alvares da; https://orcid.org/0000-0002-8078-0304; http://lattes.cnpq.br/6445492335035108; Paula, Flavia de; https://orcid.org/0000-0001-8679-2982; http://lattes.cnpq.br/7913201450663683; Tessarollo, Nayara Gusmão; http://lattes.cnpq.br/8607496356516938; Lyra Junior, Paulo Cilas Morais; https://orcid.org/0000-0002-4403-6684; http://lattes.cnpq.br/5084773867229309Epithelial ovarian cancer (EOC) is the most lethal gynecological neoplasm, characterized by high recurrence rates and resistance to cisplatin (CDDP) and PARP inhibitor treatments. Understanding the mechanisms related to these phenotypes is urgent. Therefore, in the first chapter of this study, the role of NLRP1 and NLRP3 inflammasomes and pyroptosis in CDDP chemoresistance is investigated. Cell viability of A2780, ACRP, and OVCAR3 cell lines were measured by the MTT assay, and IC50 values were determined. Cytokine levels in the supernatant were measured by ELISA. Caspase-1 activation was detected by the Caspase-Glo® 1 assay. The cell death profile was determined by flow cytometry (Annexin V/PI), and membrane pore formation was measured using PI. A cell migration assay was performed combining CDDP and the caspase-1 inhibitor. The IC50 of primary cells (PtAF) and viability after treatment with NLRP3 inhibitor (MCC950)+CDDP were determined by Cell Counting Kit-8 (CCK-8). The IC50 values indicated different CDDP resistance profiles in the cell lines. Secretion of inflammatory cytokines, caspase-1 activation, lytic cell death profile, and pore formation were higher in cells treated with CDDP. Caspase-1+CDDP inhibition reduced cell migration. PtAF treated with MCCC950+CDDP decreased cell viability and proliferation. The data suggest that inflammasome activation and pyroptosis are mechanisms associated with EOC chemoresistance to CDDP, contributing to the devastating scenario of this disease. Targeting these pathways may represent a promising treatment strategy. In the second chapter, the effect of the PARP inhibitor (Rucaparib) on mitochondrial reprogramming and its contribution to resistance is explored. Using primary cells: PtAF, PtD, and PtAD, viability assays with Rucaparib and Fludarabine Phosphate were performed by CCK-8. Oxygen consumption and extracellular acidification were detected by Seahorse. Mitochondrial content and mitochondrial membrane potential were determined by immunofluorescence with Mitotracker deepened and TMRE. NADH content was identified by autofluorescence microscopy (FLIM). The viability of cells treated with mitochondrial complex inhibitors was determined by crystal violet. The results indicated increased mitochondrial content, oxygen consumption, extracellular acidification especially in PtAF and increased NADH levels in the other cell lines. The combination of fludarabine phosphate with rucaparib had an additional cytotoxic effect on PtAF and PtD. These data suggest that PARPi affects mitochondrial metabolism and mass of cell lines differentially and that NAD transport (via SLC25A51) may play a key role in resistance, representing a potential target. Taken together, these data contribute to understanding the mechanisms underlying treatment resistance in EOC, highlighting inflammatory and mitochondrial targets as potential pathways for new therapeutic approaches.
- ItemAnálise da metilação do gene BDNF e a relação com estresse em policiais militares da Grande Vitória - ES(Universidade Federal do Espírito Santo, 2025-08-01) Quaioto, Bárbara Risse; Souza, Marcele Lorentz Mattos de; https://orcid.org/0000-0002-7364-8129; http://lattes.cnpq.br/8021072835101893; Silva, Adriana Madeira Álvares da; https://orcid.org/0000-0002-8078-0304; http://lattes.cnpq.br/6445492335035108; https://orcid.org/0000-0002-3062-7619; http://lattes.cnpq.br/7948833367111238; Errera, Flávia Imbroisi Valle; https://orcid.org/0000-0002-8069-6372; http://lattes.cnpq.br/9337327437538048; Gouvea, Sonia Alves; https://orcid.org/0000-0001-5180-471X; http://lattes.cnpq.br/7268228122543743; Rocha, José Cláudio Casali da; http://lattes.cnpq.br/6612912880886333; Arantes, Lídia Maria Rebolho Batista; https://orcid.org/0000-0001-8230-1218; http://lattes.cnpq.br/2019308149950531Stress is characterized by an imbalance in homeostasis, caused by factors known as stressors, which trigger physiological and psychological responses in the body. Repeated and high-level stress situations can alter the functioning of various bodily systems, compromising both physical and mental health. Due to the multiple adverse conditions faced by military police officers, these professionals have become a key target for scientific research. It is known that chronic stress reduces the activity of brain-derived neurotrophic factor (BDNF), which impairs several regions critical for the stress response, such as the hippocampus, prefrontal cortex, and amygdala. Moreover, this reduction is associated with the development of mental disorders. Stress can also lead to changes in epigenetic mechanisms, such as DNA methylation, thereby influencing gene expression. Therefore, this study aimed to investigate the relationship between BDNF gene methylation and stress in military police officers. To this end, 123 military police officers from the metropolitan region of Greater Vitória, Espírito Santo, Brazil, were recruited for the assessment of socioeconomic factors, health conditions, lifestyle, and stress levels. Blood samples were collected for the extraction of genomic DNA and subsequent analysis of methylation in the promoter region of exon IV of the BDNF gene. A generalized linear model was used to analyze the effect of stress on methylation levels, incorporating potential confounding factors. The results showed that stress was significantly associated with DNA methylation at CpG sites 5, 6, and 11 in exon IV of the BDNF gene, indicating that stress increases the likelihood of higher methylation levels at these sites. In light of these findings, we suggest that stress may act as an environmental pressure factor that modifies BDNF gene methylation. Additionally, the study provides support for public policies focused on stress management, especially for high-risk professionals.
- ItemEfeito Neuroprotetor da Resina de Virola oleifera na Isquemia Cerebral Induzida em Camundongos Hipercolesterolêmicos Knockout para ApoE(Universidade Federal do Espírito Santo, 2025-03-17) Peçanha, Marcos Alves de Souza; Nogueira, Breno Valentim ; https://orcid.org/0000-0002-2199-0635; http://lattes.cnpq.br/0011229320439147; https://orcid.org/0000-0002-5181-5600; http://lattes.cnpq.br/0043785257791561; Oliveira, Jairo Pinto de; https://orcid.org/0000-0001-7595-1183; http://lattes.cnpq.br/2228283301316218; Prado, Adilson Ribeiro ; https://orcid.org/0000-0001-8808-4488; http://lattes.cnpq.br/3085491325255749; Aires. Rafaela ; https://orcid.org/0000-0002-9532-9401; http://lattes.cnpq.br/5449202639864996; Larissa Zambom Côco; http://lattes.cnpq.br/6978169622337568; Larissa Zambom Côco; https://orcid.org/0000-0003-1601-8419; http://lattes.cnpq.br/6978169622337568Ischemic stroke (IS) remains a global public health challenge, affecting millions of people and impacting the quality of life of patients and their families. Characterized by complex pathophysiological mechanisms such as cerebral hypoperfusion, excitotoxicity, and oxidative stress, stroke is aggravated by hypercholesterolemia, an important factor that intensifies neuronal lesions and inflammatory processes. Given the limitations of conventional therapies, interest is growing in natural alternatives capable of offering neuroprotection. This study evaluated the neuroprotective potential of Virola oleífera resin, an Amazonian plant rich in phenolic compounds and flavonoids, in ApoE-/- mice with hypercholesterolemia submitted to global cerebral ischemia. The animals were given resin orally (300 mg/kg/day) for seven days before stroke induction. After 48 hours, behavioral, biochemical, and histological evaluations were carried out. The results revealed that treatment with Virola oleifera significantly reduced the area of cerebral infarction, improved motor and cognitive functions, and reduced the amount of necrotic neurons in the affected cortex. In addition, there was a reduction in total cholesterol, LDL, and triglyceride levels, as well as advanced protein oxidation products (AOPPs), a reduction in oxidative stress, and inflammation. These findings suggest that Virola oleifera resin acts in a multifactorial way, protecting the brain against ischemic damage, especially in conditions of metabolic risk. The data obtained reinforces the biotechnological potential of this natural resource as a basis for developing new therapeutic strategies aimed at the prevention and treatment of stroke, encouraging future translational and clinical research.
- ItemEfeitos da aplicação de estimulação transcraniana por corrente contínua e interface cérebro-computador na atividade cerebral e função física de membros inferiores de pacientes pós-AVC(Universidade Federal do Espírito Santo, 2024-08-28) Lima, Jéssica Paola Souza; Bastos Filho, Teodiano Freire ; https://orcid.org/0000-0002-1185-2773; http://lattes.cnpq.br/3761585497791105; https://orcid.org/0000-0002-6294-5179; http://lattes.cnpq.br/0393395559098385; Andrade, Adriano de Oliveira ; https://orcid.org/0000-0002-5689-6606; http://lattes.cnpq.br/1229329519982110; Espírito Santo, Caroline Cunha do ; https://orcid.org/0000-0001-8657-9532; http://lattes.cnpq.br/4920759696380516; Nogueira, Breno Valentim ; https://orcid.org/0000-0002-2199-0635; http://lattes.cnpq.br/0011229320439147; Caldeira, Eliete Maria de Oliveira ; https://orcid.org/0000-0002-3742-0952; http://lattes.cnpq.br/8508979665054143Restoring motor function after a stroke is complex, involving neuroplasticity, spontaneous recovery, and therapeutic interventions. Despite advances in motor rehabilitation for post-stroke patients, gaps remain, especially for chronic patients. This research aims to evaluate the functional evolution of a subject who suffered a stroke after applying a protocol that combines transcranial Direct Current Stimulation (tDCS) with a Brain-Computer Interface (BCI) based on Motor Imagery (MI), also integrating Virtual Reality (VR) and a Motorized Pedal (MP). Inicialmente, uma revisão sistemática da literatura foi realizada a fim de embasar cientificamente as escolhas metodológicas do estudo clínico. This study adopted the Alternating Treatment Design (ATD), with tDCS applied for 20 minutes with a current intensity of 2 mA, followed by 20 minutes of motor training with the BCI, five times a week, for two weeks. Functional and somatosensory assessments were performed weekly using standardized scales, and compared using the Friedman Test. Cortical effects were analyzed through Event-Related Desynchronization (ERD) patterns and brain connectivity changes, collected via electroencephalogram (EEG). Results showed improvements in muscle strength, range of motion, and functional capacity of the selected patient. Additionally, EEG analysis revealed increased neural activity with the combined use of tDCS and BCI compared to Baseline and Sham. Continuing research in this field is strongly recommended, exploring different stimulation parameters, patient populations, and emerging technologies, such as functional neuroimaging
- ItemAnálise da diversidade genética e identificação de citros (Citrus spp) com marcadores moleculares e High Resolution Melting (HRM)(Universidade Federal do Espírito Santo, 2024-12-12) Britto, Karolinni Bianchi; Paneto, Greiciane Gaburro; https://orcid.org/0000-0001-8035-4199; http://lattes.cnpq.br/8176374147579841; Ventura, José Aires ; https://orcid.org/0000-0003-1422-1739; http://lattes.cnpq.br/8687116881326074; https://orcid.org/0000-0002-1614-0898; http://lattes.cnpq.br/3604508015056223; Guimarães, Marco Cesar Cunegundes ; https://orcid.org/0000-0003-2146-0180; http://lattes.cnpq.br/0261991057482057; Oliveira, Jairo Pinto de ; https://orcid.org/0000-0001-7595-1183; http://lattes.cnpq.br/2228283301316218; Ferreira, Marcia Flores da Silva ; https://orcid.org/0000-0003-1541-6634; http://lattes.cnpq.br/5719813884063445; Costa, Helcio; https://orcid.org/0000-0002-7056-9927; http://lattes.cnpq.br/2086091514383462Citrus farming plays a crucial role in national fruit production, encompassing the cultivation of several species and varieties of the Citrus genus. Citrus growers face challenges related to seedling certification, due to the similarity between the leaves of the cultivars and the possibility of exchange between them, since the morphological characteristics manifest themselves in a subtle and variable way. The selection of molecular markers for citrus is a challenge due to the low molecular variability observed. Thus, the objective of this study was to identify Single Nucleotide Polymorphisms (SNPs) through DArTSeq technology, which combines the Diversity Arrays Technology marker system with next-generation sequencing technology, and from these markers, develop rapid identification kits using the High Resolution Melting (HRM) technique, which is sensitive and low-cost. The methodology, results and discussion were written in three chapters. The first chapter presents a review “Beyond the peel: exploring Citrus diversity through DArTSeq”. The study used DArTSeq technology to analyze genetic variation in 93 accessions of nine citrus species, resulting in the identification of 64,442 SNPs and 69,963 SilicoDArT markers, which after filtering were reduced to 9,073 and 3,496, respectively. The various analyses revealed eight clusters and six genetic groups. The research also established an important basis for future investigations in citrus genetics. The second chapter presents a patent filed with the National Institute of Industrial Property (INPI), entitled “Oligonucleotide primers and method for genetic discrimination and certification of Pera orange (Citrus sinensis) cultivars of agricultural interest using the High Resolution Melting (HRM) technique”. A kit, with seven pairs of primers for HRM analysis, was developed to identify eight Pera orange cultivars. The third chapter presents a patent filed with the INPI, entitled “Oligonucleotide primers and method for genetic discrimination and certification of citrus rootstocks (Family Rutacea) of agricultural interest using the High Resolution Melting (HRM) technique”. A kit with three pairs of primers for HRM analysis was developed to identify seven genotypes of citrus rootstocks, five species and two hybrids. This thesis contributes to analyses of the genetic diversity and population structure of citrus, reflecting the relationships both between and within species. The identification of the cultivars was performed using oligonucleotide primer kits using the HRM technique. In addition, these kits developed will assist in the rapid identification of Pera orange cultivars and citrus rootstocks by HRM, and can be implemented in certification. Thus, the proposed markers proved to be effective in discriminating citrus genotypes, overcoming the limitations of morphological identification, especially during the initial phases of the plant, offering a significant advance for the citrus sector and promoting greater security for producers by ensuring the authenticity of the propagation material