Doutorado em Biotecnologia RENORBIO
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- 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
- ItemNanopartículas superparamagnéticas de óxido de ferro : estratégias de síntese e funcionalização para adsorção de proteínas e detecção de anticorpos(Universidade Federal do Espírito Santo, 2025-02-21) Keijok, Wanderson Juvencio; Oliveira, Jairo Pinto de ; https://orcid.org/0000-0001-7595-1183; http://lattes.cnpq.br/2228283301316218; Guimarães, Marco Cesar Cunegundes ; https://orcid.org/0000-0003-2146-0180; http://lattes.cnpq.br/0261991057482057; https://orcid.org/0000-0001-6352-7182; http://lattes.cnpq.br/7880904848142144; Nogueira, Breno Valentim ; https://orcid.org/0000-0002-2199-0635; http://lattes.cnpq.br/0011229320439147; Miguel, Emilio de Castro ; https://orcid.org/0000-0003-4130-672X; http://lattes.cnpq.br/0690009743607948; Rossi, André Linhares ; https://orcid.org/0000-0002-9920-4411; http://lattes.cnpq.br/0700787558627219A detecção rápida e precisa de biomoléculas, como proteínas e anticorpos, é crucial para o avanço da biotecnologia para o diagnóstico de doenças infecciosas. Neste contexto, as nanopartículas de óxido de ferro superparamagnéticas (SPIONs) são plataformas promissoras devido às suas propriedades únicas, como alta área de superfície, superparamagnetismo e facilidade de funcionalização. Este trabalho explora o uso de SPIONs em duas aplicações distintas, demonstrando seu potencial em biotecnologia. No primeiro capítulo, desenvolvemos SPIONs estabilizadas com tetraetoxissilano (TEOS) para a adsorção eficiente de albumina de soro bovino (BSA). Utilizando planejamento fatorial e metodologia de superfície de resposta (RSM), foi sintetizada nanopartículas com diâmetro médio de 11,06 ± 0,84 nm e capacidade de adsorção de 87,8 ± 1,79 mg de BSA por grama. As SPIONs demonstraram alta estabilidade coloidal e potencial zeta de -46,24 mV, eficazes em pH de 2 a 10 e concentrações salinas de até 1M, tornando-as ideais para a imobilização de biomoléculas. No segundo estudo, investigamos a funcionalidade das SPIONs com a proteína Spike do SARS-CoV-2 para a detecção de anticorpos IgG no sangue humano. As SPIONs foram sintetizadas por coprecipitação e funcionalizadas com (3-aminopropil) trietoxissilano (APTS), permitindo uma conjugação eficiente da proteína Spike. A caracterização por FTIR, espectroscopia Raman e difração de raios X confirmou a presença de PEG, APTS e integridade cristalina. A capacidade de adsorção máxima da proteína Spike foi de 8 ± 1,14 mg por 20 mg de SPIONs. O sensor demonstrou alta sensibilidade, com limite de detecção de 0,5 ng/mL para IgG, coletado ao ELISA. A validação com amostras de sangue humano mostrou alta sensibilidade e especificidade na detecção precoce de infecção. As SPIONs desenvolvidas demonstraram alta eficiência na adsorção de biomoléculas e sensibilidade na detecção de anticorpos, evidenciando seu potencial para diagnósticos clínicos de alta precisão. Os resultados obtidos reforçam a viabilidade dessas nanopartículas como plataformas promissoras para biossensores e outras aplicações biomédicas, abrindo caminho para avanços na detecção de doenças infecciosas
- ItemDesenvolvimento de coração bioartificial a partir de arcabouço de matriz extracelular descelularizada enriquecida com fibronectina plasmática humana(Universidade Federal do Espírito Santo, 2023-02-27) Taufner, Gabriel Henrique; Nogueira, Breno Valentim; https://orcid.org/0000000221990635; http://lattes.cnpq.br/0011229320439147; https://orcid.org/0000-0002-4040-5948; http://lattes.cnpq.br/4425677222076255; Baldo, Marcelo Perim; https://orcid.org/0000-0002-7673-3580; http://lattes.cnpq.br/7820422119282248; Oliveira, Jairo Pinto de; https://orcid.org/0000000175951183; http://lattes.cnpq.br/2228283301316218; Prado, Adilson Ribeiro; https://orcid.org/0000-0001-8808-4488; http://lattes.cnpq.br/3085491325255749; Diaz, Camilo Arturo Rodriguez; https://orcid.org/0000000196575076; http://lattes.cnpq.br/2410092083336272The reconstruction of complex organs after decellularization presents as the greatest challenge the delivery, adhesion, proliferation and differentiation of cells. To succeed in the process, it is necessary to mimic the in vivo microenvironment as much as possible. It is known that decellularization is capable of removing important biomolecules in the morphophysiology of the organ, such as glycosaminoglycans and glycoproteins such as fibronectin. Fibronectin remaining in the extracellular matrix after the decellularization process may not be sufficient to satisfactorily promote the tissue reconstruction process, especially if the scaffold is in the adult stage of development, where the composition of said glycoprotein is known to be reduced. In our study, we obtained decellularized scaffolds to investigate the influence of human plasma fibronectin on adult mouse heart reconstruction. We demonstrated that the decellularized scaffolds were considerably preserved in terms of their matrisomal biomolecular composition. We emphasize the maintenance of the hydroxyproline molecule, found in a concentration of 5,509 ± 818.13 vs. 4,881 ± 1,487 µg/total dry weight in decellularized adult organ and control respectively. Regarding neonatal decellularized scaffolds, we observed maintenance of hydroxyproline when compared to its native control (330.5 ± 17.60 vs. 273.9 ± 12.30 µg/total dry weight). In addition to matrisomal biomolecules, we emphasize the remarkable reduction of DNA (83.63% and 93% in neonatal and adult respectively) and residual SDS (33.92% and 96.44% in neonatal and adult respectively), interfering with the reconstruction process . We investigated the use of a non-destructive analytical approach for decellularized tissues: Raman spectroscopy, whose results corroborated the spectrophotometric analyzes of matrisomal biomolecules. We established an effective strategy for the recomposition of fibronectin in adult decellularized scaffolds, however, we found that despite the success in the recomposition of the glycoprotein, we did not find statistical significance after the reconstruction of the organs from cells of the H9c2 lineage. Given this last finding, we suggest additional studies capable of investigating other classes of fibronectin. Furthermore, it is also necessary to investigate cell populations beyond the H9c2 lineage.
- ItemPadrões alimentares processados e ultraprocessados e a relação com a metilação dos genes BDNF e NR3C1 e o estado nutricional(Universidade Federal do Espírito Santo, 2023-02-24) Vieira, Tamires dos Santos; Silva, Adriana Madeira Alvares da; https://orcid.org/0000000280780304; http://lattes.cnpq.br/6445492335035108; https://orcid.org/0000000238993664; http://lattes.cnpq.br/9014510901567961; Nogueira, Breno Valentim; https://orcid.org/0000000221990635; http://lattes.cnpq.br/0011229320439147; Hollais, André Willian; https://orcid.org/0000-0002-2991-8646; http://lattes.cnpq.br/0068789063590867; Gouvea, Sonia Alves; https://orcid.org/000000015180471X; http://lattes.cnpq.br/7268228122543743The change in the lifestyle of the population with the acquisition of unhealthy habits, such as the industrialized diet, have contributed to the growth of obesity, increase in the prevalence of diabetes, hypertension and other non-transmissible chronic diseases such as depression and cancer. Diet composition can act directly as a transcription factor, in addition to influencing epigenetic modifications, altering gene expression and contributing to the risk of diseases which can be transferred to offspring. Thus, the aim of this study was to evaluate the nutritional status and the relationship between adherence to processed and ultra-processed dietary patterns and levels of methylation of the BDNF and NR3C1 genes. This cross-sectional study collected data from 250 individuals between 20 and 59 years of age, using as indicators the Body Mass Index (BMI), HDL-C and LDL-C for dyslipidemia, TyG index for insulin resistance and symptoms suggestive of depression by the Beck Depression Inventory. The independent variables were socioeconomic factors (gender, age, education, location and income), psychosocial factors (stress, anxiety, depression and food insecurity), nutritional status (TyG Index, HDL-c, LDL-c, BMI, cortisol) and lifestyle (physical activity, smoking and alcoholism) used to adjust the generalized linear model (GLzM). The BDNF and NR3C1 genes methylation profile was evaluated by the pyrosequencing method. Dietary patterns were defined by factor analysis that assessed the interrelationship between commonly consumed foods, resulting in three patterns: healthy, industrialized and mixed. For the methylation of the BDNF sitespecific CpGs, it was decided to work separately, while for the NR3C1 CpGs, the main components were extracted, obtaining two components of CpGs. Comparisons of medians, Spearman correlations and linear models were used to assess the association between BDNF methylation and the 1F region of the NR3C1 gene. The prevalence of overweight and obesity was 33.5% and 39.9%, respectively. Furthermore, 64.9% of the individuals had insulin resistance, and more than 70.0% had an increased risk for metabolic complications related to abdominal adiposity, waistto-height ratio and conicity index. The present study showed that adherence to an unhealthy eating pattern and excess weight played a significant role in BDNF gene methylation, increasing the percentage of methylation (p<0.05). In addition, the lower methylation in the total segment and in component 1 (CpG 44-47) of NR3C1 can be explained by overweight and adherence to the industrialized pattern, when controlled for covariates. And this evidence helps to understand the epigenetic role of adherence.