Doutorado em Biotecnologia

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    Uma nova proposta para terapia psicomotora e método de avaliação de crianças e adolescentes com síndrome de Down e com transtorno do espectro autista utilizando robô socialmente assistivo
    (Universidade Federal do Espírito Santo, 2024-10-25) Schreider, Sheila da Luz; Caldeira, Eliete Maria de Oliveira; Bastos Filho, Teodiano Freire; https://orcid.org/0000-0002-1185-2773; Gouvêa, Sônia Alves; Jiménez, Nicolás Jacobo Valencia; Freitas, Éberte Valter da Silva; Santo, Caroline do Espírito
    Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by atypical neuropsychomotor development, communication and social interaction impairments, repetitive behaviors, stereotypies, and a preference for a restricted range of interests and activities. Children with ASD may present with muscle hypotonia and balance deficits in early childhood, leading to neuropsychomotor delay. Down Syndrome (DS) is the most common genetic alteration in humans, characterized by an imbalance in chromosomal constitution related to chromosome 21. Hypotonia is a typical feature in individuals with DS, influencing postural control and proprioception. This study aims to evaluate the effects of three therapeutic protocols of game therapy, using serious games applied through the socially assistive robot MARIA T-21, which stands for "Mobile Autonomous Robot for Interaction with Autistics and Trisomy 21 (Down Syndrome)," to assess proprioception and postural balance in children and adolescents with ASD and DS. Additionally, the study evaluated the functional performance and therapeutic progress of these children and adolescents through body image analysis, captured by a multi-camera system placed in the four corners of the experimental room. The serious games used in the research were designed by the MARIA T-21 robot. The sample consisted of 20 children and adolescents, with 11 having ASD and 9 with DS, aged between 5 and 18 years, divided into three data collection groups. These children and adolescents were recruited from institutions such as the Associação dos Amigos dos Autistas do Espírito Santo (AMAES Vitória), Vitória Down, and Associação de Pais e Amigos dos Excepcionais (APAE) in Vitória and Vila Velha. Before starting the serious game protocols, parents/guardians were informed about the study objectives, and the participants underwent a psychomotor assessment based on Victor da Fonseca’s Psychomotor Battery. Parents/guardians of children and adolescents in the third data collection phase answered the PEDI-CAT questionnaire, a functional assessment tool, before and after the application of the serious game protocol. This protocol consisted of 10 sessions of 45 minutes at AMAES Vitória and APAE Vitória and 4 sessions of 45 minutes at APAE Vila Velha, held twice a week, focusing on proprioception, balance, and both gross and fine praxia. The games used included Sound Sequence, Arara Game, Hopscotch, Healthy Food, Cross Kids, Bricks, and Star Wars, along with Warm-Up and Drawing activities (called Draw with Me). At the end of the protocol, participants underwent a psychomotor assessment identical to the initial one. The results showed that the use of the MARIA T-21 robot and serious games was an effective therapeutic strategy, providing a playful approach and increasing children's adherence to therapy. There was a significant improvement in psychomotor profile, with gains in posture maintenance, such as unipedal support and tiptoe standing, as well as an increase in average levels achieved in the Arara Game. Body image analysis of two adolescents with DS revealed fluctuations in postural control, with improved motor efficiency over the sessions. The image analysis tool used, BalancePro, proved highly valuable for tracking the progress of these two evaluated adolescents, providing numerical data on movement during the analyzed task. In conclusion, the use of the MARIA T-21 robot and serious games in this study proved to be a promising alternative for motor rehabilitation in children and adolescents with ASD and DS, contributing to improved motor skills and overall development, facilitating their integration into school and social settings, and promoting greater autonomy. It is believed that, for families, these advances may lead to improved quality of life and family interactions, as well as facilitate the social and school inclusion of children and adolescents.
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    Evaluating the effect of the simultaneous cerebrospinal stimulation, motor imagery, virtual reality and pedaling on post-stroke patients
    (Universidade Federal do Espírito Santo, 2024-09-16) Mehrpour, Sheida; Andrade, Adriano de Oliveira ; https://orcid.org/0000-0002-5689-6606; http://lattes.cnpq.br/1229329519982110; Bastos Filho, Teodiano Freire ; https://orcid.org/0000-0002-1185-2773; http://lattes.cnpq.br/3761585497791105; https://orcid.org/0000-0002-1217-8071; http://lattes.cnpq.br/4006017652838495; Espírito Santo, Caroline Cunha; https://orcid.org/0000-0001-8657-9532; http://lattes.cnpq.br/4920759696380516; Rodríguez, Denis Delisle ; https://orcid.org/0000-0002-8937-031X; http://lattes.cnpq.br/7140331839822423 ; Fernandez, Antônio Alberto Ribeiro ; https://orcid.org/0000-0003-0535-9349; http://lattes.cnpq.br/4696507759154477; Nogueira, Breno Valentim ; https://orcid.org/0000-0002-2199-0635; http://lattes.cnpq.br/0011229320439147
    Technology in medicine is transforming the healthcare landscape by enhancing diagnostics, treatment, and patient management. With the integration of advanced tools and systems, healthcare professionals can deliver more accurate and timely care. Innovations such as telemedicine, artificial intelligence, and electronic health records streamline processes and improve communication among providers. Additionally, technology facilitates personalized medicine, allowing treatments to be tailored to individual patients based on their unique needs. The ongoing evolution of medical technology not only increases efficiency but also expands access to healthcare, ensuring that patients receive the best possible outcomes. As technology continues to advance, its role in medicine will become even more pivotal in shaping the future of healthcare. Stroke is the leading cause of acquired physical disability in humans, and the second largest cause of global mortality. Technology in stroke rehabilitation plays a vital role in enhancing recovery outcomes for patients. Advanced tools such as virtual reality, robotics, Brain-Computer Interface based on Motor Imagery (BCI-MI), Non Invasive Brain Stimulation (NIBS) techniques, and telehealth platforms offer innovative ways to engage patients in their rehabilitation process. Virtual reality can simulate real life scenarios, helping patients practice daily activities in a safe environment, while robotic exoskeletons assist in retraining motor functions through repetitive movements. Telehealth enables remote therapy sessions, providing continuous support and flexibility for patients to engage in their recovery from home. Additionally, wearable devices allow for real-time monitoring of progress, ensuring that treatment plans can be adjusted to meet individual needs effectively. Overall, these technological advancements are reshaping stroke rehabilitation, making it more personalized, accessible, and efficient. Non-Invasive Brain Stimulation (NIBS) techniques, such as transcranial direct current stimulation (tDCS) and transcutaneous spinal Direct Current Stimulation (tsDCS), are increasingly being applied in stroke rehabilitation to enhance recovery outcomes. These methods work by modulating neuronal activity in targeted brain regions, promoting neuroplasticity and facilitating motor function recovery. By improving communication between brain areas affected by the stroke and those responsible for movement, NIBS can help patients regain lost skills more effectively. As research continues to advance, these techniques hold promise for optimizing rehabilitation strategies and improving the quality of life for stroke survivors. The main objective of this study is to develop new, low-cost rehabilitation methods to patients with subacute to chronic stroke, aiming to increase neuroplasticity and improve motor function through combining methods such as tDCS plus tsDCS, VR, MI and pedaling exercise. This research are divided into three separate Chapters to assess both the long-term effects (Chapter I) and the immediate effects (Chapters II and III) of the intervention. In chapter I, the study was set up with the Alternative Treatment Design (ATD), comprising three phases: baseline, sham stimulation, and real stimulation. For Chapters II and III, the study design was defined as a pre- and post-stimulation assessment. For the experiment in the first Chapter, four subacute hemiparetic stroke patients were selected. The same experiment and participants were recruited for Chapters two and three, but the methodology for evaluating the effects of the intervention differed between these Chapters. For Chapters two and three, a total of eight participants were selected, including four patients and four healthy individuals. In both experiments, participants were randomly assigned to two groups to receive cerebrospinal stimulation, according to two different protocols (conventional and periodic). Participants in the conventional stimulation group received 20 minutes of stimulation, while those in the periodic stimulation group underwent two 13-minute stimulation sessions separated by a 20-minute rest period. The anode electrode was placed over the M1 region of the affected hemisphere, guided by the 10/20 International System. The cathode electrode was positioned centrally on the spinous process of the thoracic vertebra at T11 (T10-T12) by palpation. For the first experiment the results were evaluated using surface electromyography (sEMG), Maximum Voluntary Contraction (MVC), Fugl-Meyer Assessment for Lower Extremity (FMA-LE), miniBESTest, goniometry, 10-meter walk test (10MWT), pedaling speed, as well as specific stroke scales. In the second experiment, in addition to stimulation, Virtual Reality was used to enhance Motor Imagery (MI) effect in order to evaluate the combined effect on Mu and Beta bands modulation in post-stroke patients and healthy individuals. Results from the second experiment were analyzed using quantitative electroencephalography (EEG) measures, such as cortical topography based on mean amplitude values, brain connectivity parameters such as Phase Locking Value (PLV) and Magnitude Squared Coherence (MSC). For Chapter III, the Hjorth parameters (activity, mobility, complexity) were used across two assessment sessions, pre- and post-stimulation. The results of the experiment presented in Chapter I indicated significant improvements in muscle contraction, motor function and gait among patients. Participants in the conventional stimulation protocol group showed enhancements in tibialis muscle contraction, as assessed by sEMG and ankle dorsiflexion goniometry. In contrast, those in the periodic stimulation protocol exhibited improvements in motor function measures such as FML-LE, MiniBestest, and the 10- meter walk. Findings from the Chapter II experiment revealed different patterns of brain connectivity under the combined effects of cerebello-spinal stimulation, along with VR and MI, in both patients and healthy controls, emphasizing the need for personalized treatments for post-stroke patients. Results of Chapter III showed that the beta band is more sensitive to modulation by the combined methods compared to the Mu band, which was more reactive in patients than in healthy controls. The Activity parameter had a greater influence on the modulation of Mu and Beta bands in both patients and healthy controls. While the Mobility parameter showed greater influence in patients, the Complexity parameter was more sensitive in healthy controls. Due to the variability of the results and the small sample size, it was challenging to distinguish the effects of the two stimulation protocols in Chapters II and III
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    Herbicida à base de glifosato como potencial fator de risco para o câncer de mama: uma análise da expressão gênica, das modificações epigenéticas e do uso de epifármacos
    (Universidade Federal do Espírito Santo, 2024-08-26) Alves, Lyvia Neves Rebello; Santos, Eldamária de Vargas Wolfgramm dos; Louro, Iúri Drumond; https://orcid.org/0000-0001-5160-9615; https://orcid.org/0000-0001-5107-3689; Paula, Flávia de; Batitucci, Maria do Carmo Pimentel; Pereira, Fausto Edmundo Lima; Carvalho, Elizeu Fagundes de
    Breast cancer is the most common neoplasm in women worldwide, with both genetic and environmental factors playing a role in its development. Glyphosate, the active ingredient in widely used agricultural herbicides, is recognized as a potential carcinogen and endocrine disruptor, making it a candidate for inducing epigenetic modifications linked to breast cancer. This study investigates the effects of the glyphosate-based herbicide Roundup® on non-tumorigenic (MCF10A) and tumorigenic (MCF7 and MDA-MB-231) breast cell lines, focusing on the expression of key breast cancer-related genes. Additionally, the study examines the association with epigenetic modifications and the use of epidrugs (5-Aza-2′-deoxycytidine, 3-Deazaneplanocin A, and Trichostatin A) to reverse potential alterations, aiming to understand the risks and mechanisms of herbicide action. Results indicate that Roundup® affects cells through a non-estrogenic mechanism, impacting both hormone-dependent and -independent cell lines with varying toxic and proliferative effects depending on dose and exposure time. Moreover, it altered the expression of breast cancer-related genes such as BRCA1 and BRCA2 at low doses. The use of epigenetic modulators was able to reverse some Roundup®-induced changes, suggesting the herbicide's role in epigenetic modifications. Overall, these findings highlight the importance of understanding glyphosate-based herbicide mechanisms in humans, which could enable personalized prevention strategies to mitigate breast cancer risks.
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    Determinação do perfil molecular dos extratos de folhas de Carica papaya (L.) em busca de biomarcadores estágio-dependentes relacionados à doença da meleira do mamoeiro
    (Universidade Federal do Espírito Santo, 2024-07-12) Britto, Isabella Oliveira; Santos, Alexandre Martins Costa; https://orcid.org/0000-0002-8801-8875; http://lattes.cnpq.br/; https://orcid.org/; http://lattes.cnpq.br/; Leite, João Paulo Viana; https://orcid.org/; http://lattes.cnpq.br/; Silva, Diolina Moura ; https://orcid.org/; http://lattes.cnpq.br/; Gonçalves, Juliana Barbosa Coitinho; https://orcid.org/; http://lattes.cnpq.br/; Lima, Graziela Domingues de Almeida; http://lattes.cnpq.br/
    Papaya Sticky Disease is caused by PMeV viral complex and poses a significant threat to papaya production worldwide. Infected plants remain asymptomatic until flowering and fructification, acting as silent reservoirs of the virus in the field. Secondary metabolites could act as potential biomarkers of disease progression and can be detected using chromatographic techniques for early disease diagnosis. In this study, molecular profiles of ethanolic extracts from C. papaya leaves in pre- and postflowering (2.5, 5.0, and 7.5 mg.mL-1) and fractions were evaluated by UV-Vis spectroscopy, reversed-phase chromatography, ion exchange chromatography, and mass spectrometry. The 7.5 mg.mL-1 concentration was selected as the experimental concentration for crude extracts. Chromatograms showed symmetrical peaks, eluted within similar retention time ranges across the three plant stages, with narrow base widths, similar peak shapes, and no tailing, suggesting the presence of a limited number of potentially isolable compounds. Analytical parameters indicated quantitatively larger chromatographic peaks in pre-flowering leaf extracts compared to post-flowering ones, reflecting a possible defense response against viral infection. Significant differences in chromatographic profiles between the different plant stages were observed, indicating the potential of these groups as biomarkers for Papaya Sticky Disease. This study developed a sensitive and reproducible chromatographic method to distinguish plant stages, identifying flavonoids and alkaloids as components of strategic groups related to the defense system against viral infections.
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    Potencial prognóstico de células inflamatórias e PD-L1 solúvel em carcinoma epidermoide de cabeça e pescoço
    (Universidade Federal do Espírito Santo, 2024-04-16) Daniel, Camila Batista; 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; Zeidler, Sandra Lúcia Ventorin von ; https://orcid.org/0000-0002-8897-5747; http://lattes.cnpq.br/5785612863130498; Orientador2; https://orcid.org/; http://lattes.cnpq.br/; https://orcid.org/; http://lattes.cnpq.br/; Santos, Eldamária de Vargas Wolfgramm dos ; https://orcid.org/; http://lattes.cnpq.br/; Errera, Flavia Imbroisi Valle ; https://orcid.org/; http://lattes.cnpq.br/; Arantes, Lidia Maria Rebolho Batista ; https://orcid.org/; http://lattes.cnpq.br/; Mendonça, Elismauro Francisco de ; 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/
    Immunosuppression is recognized as a hallmark of cancer and has been associated with worse outcomes. In head and neck squamous cell carcinoma (HNSCC), immunosuppression is a hallmark of the tumour and may be mediated by immunosuppressive inflammatory cells and PD-L1 expression. Given their participation in this process, our study aimed to describe the prognostic impact of these elements in tumour tissue and peripheral blood HNSCC patients. We conducted a multicentre retrospective study with biological samples and clinical data from HNSCC patients recruited at two centres in Brazil and the United Kingdom and healthy individuals. To analyse the inflammatory infiltrate, we used tumour tissue slides stained with haematoxylin and eosin and immunohistochemistry for PD-L1 analysis and neutrophil quantification. Absolute leukocyte counts were retrieved from pretreatment blood counts available in medical records. To evaluate the prognostic value of PD-L1 in liquid biopsy, we used serum and plasma samples obtained from patients and healthy individuals for quantification of soluble PD-L1 (sPD-L1) by ELISA. Fresh tumour tissue samples were used to analyse CD274 gene expression levels using the RT-qPCR technique. Furthermore, we analysed PD-L1 expression by flow cytometry in HNSCC cell lines and quantified sPD-L1 levels in the supernatant. The tumour microenvironment analysis showed that low levels of lymphocytes are associated with worse overall survival and disease-free survival. However, we did not observe this relationship with tumour-associated neutrophils and tumour PD-L1. When analysed together in a scoring system, we demonstrated that low levels of lymphocytes, high expression of PD-L1 and high infiltration of neutrophils are associated with a worse outcome. In blood, a high ratio of neutrophils and lymphocytes (NLR) was also associated with worse survival but was not correlated with inflammatory components of the tumour. High levels of sPD-L1 protein were associated with reduced overall survival, however our study did not identify a relationship between sPD-L1 levels in the blood and PD-L1 expression in tumour tissue, determined by immunohistochemistry and RT-qPCR. In contrast, the in vitro study showed that the levels of sPD-L1 released 14 into the supernatant are strongly correlated with cytoplasmic and membrane expression. Our data further showed that sPD-L1 levels in patients were positively correlated with the number of leukocytes, neutrophils and monocytes in peripheral blood. Taken together, our results highlight the prognostic potential of markers of the tumour microenvironment analysed in a combined manner, in a scoring system, as a way of providing a more comprehensive overview of tumour behaviour, highlighting important events, such as immunosuppression. Our results highlight the prognostic potential of PD-L1 detected by liquid biopsy in HNSCC and indicate that the levels detected in the blood do not necessarily correspond to what is observed in the tumour. Therefore, it is believed that it should be analysed as an independent marker whose immunosuppressive role is carried out at a systemic level