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- ItemRegulación de plantas genéticamente editadas y selección del gen SAMT para la edición de carica papaya buscando resistencia a meleira(Universidade Federal do Espírito Santo, 2024-05-09) Gaitán, Julia Bibiana Merchán; Rodrigues, Silas Pessini; Fernandes, Patrícia Machado Bueno; https://orcid.org/0000-0003-2695-3638; Ventura, José Aires; Tarazi, Roberto; Pérez Brito, Daisy De La Caridad; Rodrigues, Juliany Cola FernandesThis paper addresses the regulation of one of the most modern technologies for the control of plant pathologies, the CRISPR (Clustered regularly interspaced short palindromic repeats) technique, which is based on the action of a Cas protein and an gRNA that together can target specific areas of the DNA to modify them, through a systematic review of the regulatory systems of different countries. The different approaches adopted by countries in the Latin American region to regulate CRISPR technology were analyzed, taking into account the opportunities it offers and the possible associated ethical and social risks. So far, Brazil has analyzed the case-by case evaluation process of genetically modified crops by the regulatory committee CTNBio for the use of new plant breeding technologies to determine whether it would be considered a GMO or conventional breeding. This analysis was carried out because there is great interest in using CRISPR technology to develop new plant cultivars, especially plants relevant to Brazilian fruit growing. The Carica papaya cultivar, is of particular interest because of limitations in its production, due to the plant's susceptibility to the virus complex that causes papaya meleira disease, is economically and biologically relevant, as viruses have been identified in plant latex, a fluid derived from laticifers, which occur in more than 20 plant families and on which there is a body of information that points to laticifers as plant response systems to viruses. For this reason, a detailed review was conducted on the role of laticiferans of different species, examining the ability of both pro- and antiviral latex to protect plants, as well as its potential to be used as a natural source of defense molecules against insect vectors and fungal infections. The potential use of CRISPR in response to control viruses affecting laticifers was also evaluated through a compilation of articles targeting gene editing with specific expression or related to laticifer biology, confirming that it is possible to modulate laticifer function using CRISPR. The relationship of the PMeV complex in the C. papaya L. plant was discussed and the importance of the cloning of the Salicylate O-methyltransferase gene in the defense against this viral complex are discussed, due to its presence in the plant defense mechanisms related to laticifers. Describing experimental advances in the development of a protocol to silence the salicylate O-methyltransferase gene of C. papaya by means of the Golden Gate Assembly, detailing the design of gRNA for the construction and multiplication of gene editing vectors, intermediate steps in the process of gene silencing of the SAMT gene for the control of meleira disease in papaya, resulting in optimal conditions for the use of CRISPR technology in C. papaya plants.
- ItemDesenvolvimento de biomaterial ósseo descelularizado enriquecido com hidrogel de osso fetal bovino(Universidade Federal do Espírito Santo, 2024-10-10) Barros, Rodolpho José da Silva; Nogueira, Breno Valentim ; https://orcid.org/0000-0002-2199-0635; http://lattes.cnpq.br/0011229320439147; https://orcid.org/0000-0001-6214-733X; http://lattes.cnpq.br/5652361736715531; Maranduba, Carlos Magno da Costa ; https://orcid.org/0000-0001-7327-1934; http://lattes.cnpq.br/4763153859701731; 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; Aires, Rafaela; https://orcid.org/0000-0002-9532-9401; http://lattes.cnpq.br/5449202639864996The global bone graft material market is projected to grow at a rate of 6-7% annually until 2032. Conventional bone grafting strategies are no longer considered sufficient or adequate for most treatments. The development of decellularized biomaterials derived from animal tissues (xenografts) offers a promising and viable strategy. The structural similarity between animal and human bones, along with the decellularization technique, can result in a high-quality biomaterial that provides mechanical support, preserves the organic matrix, and supplies essential biological stimuli for tissue regeneration. Adult and fetal bovine bones were decellularized to produce biomaterials (blocks, granules, and hydrogel) with residual DNA levels lower than 50 ng/mg and SDS levels below 2 μg/mg of dry tissue. Thermogravimetric assays (DSC and TGA) and infrared spectroscopy (FTIR and Raman) demonstrated the adequate preservation of both the mineral and organic bone matrices. Scanning electron microscopy (SEM) and elemental composition analysis (EDS and CHNS-O) confirmed the preservation of the microstructure and biochemical composition of the bone matrix. pH stability, absence of heavy metals, and sterility were verified. Comparative tests showed that the decellularized biomaterials, particularly those derived from fetal bone, exhibit superior porosity, density, mechanical support capacity, and organic composition compared to other biomaterials. In vitro tests revealed no cytotoxicity and a higher ability to stimulate the proliferation of mesenchymal cells and fibroblasts. The results indicate that the production of decellularized bovine bone biomaterials coated with fetal hydrogel could contribute to better biocompatibility of implants and help meet the global demand for bone graft materials
- ItemUma 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 EspíritoAutism 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.
- ItemEvaluating 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/0011229320439147Technology 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
- ItemHerbicida 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 deBreast 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.