Efeitos da sinalização por Sonic Hedgehog sobre a proliferação de células-tronco neurais e gliogênese no córtex cerebral em desenvolvimento

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES

Sonic hedgehog signaling regulates mode of cell division of early cerebral cortex progenitors and increases

The morphogen Sonic Hedgehog (SHH) plays a critical role in the development of different tissues. In the central nervous system, SHH is well known to contribute to the patterning of the spinal cord and separation of the brain hemispheres. In addition, it has recently been shown that SHH signaling also contributes to the patterning of the telencephalon and establishment of adult neurogenic niches. In this work, we investigated whether SHH signaling influences the behavior of neural progenitors isolated from the dorsal telencephalon, which generate excitatory neurons and macroglial cells in vitro. We observed that SHH increases proliferation of cortical progenitors and generation of astrocytes, whereas blocking SHH signaling with cyclopamine has opposite effects. In both cases, generation of neurons did not seem to be affected. However, cell survival was broadly affected by blockade of SHH signaling. SHH effects were related to three different cell phenomena: mode of cell division, cell cycle length and cell growth. Together, our data in vitro demonstrate that SHH signaling controls cell behaviors that are important for proliferation of cerebral cortex progenitors, as well as differentiation and survival of neurons and astroglial cells.

Análise da estabilidade genética de células-tronco mesenquimais humanas

Human multipotent mesenchymal stromal cells (MSCs), also known as mesenchymal stem cells, have become an important and attractive therapeutic tool since they are easily isolated and cultured, have in vitro expansion potential, substantial plasticity and secrete bioactive molecules that exert trophic effects. The human umbilical cord as a cell source for cell therapy will help to avoid several ethical, political, religious and technical issues. One of the main issues with SC lines from different sources, mainly those of embryonic origin, is the possibility of chromosomal alterations and genomic instability during in vitro expansion. Cells isolated from one umbilical cord exhibited a rare balanced paracentric inversion, likely a cytogenetic constitutional alteration, karyotype: 46,XY,inv(3)(p13p25~26). Important genes related to cancer predisposition and others involved in DNA repair are located in 3p25~26. Titanium is an excellent biomaterial for bone-implant integration; however, the use can result in the generation of particulate debris that can accumulate in the tissues adjacent to the prosthesis, in the local bone marrow, in the lymph nodes, liver and spleen. Subsequently may elicit important biological responses that aren´t well studied. In this work, we have studied the genetic stability of MSC isolated from the umbilical cord vein during in vitro expansion, after the cryopreservation, and under different concentrations and time of exposition to titanium microparticles. Cells were isolated, in vitro expanded, demonstrated capacity for osteogenic, adipogenic and chondrogenic differentiation and were evaluated using flow cytometry, so they met the minimum requirements for characterization as MSCs. The cells were expanded under different concentrations and time of exposition to titanium microparticles. The genetic stability of MSCs was assessed by cytogenetic analysis, fluorescence in situ hybridization (FISH) and analysis of micronucleus and other nuclear alterations (CBMN). The cells were able to internalize the titanium microparticles, but MSCs preserve their morphology, differentiation capacity and surface marker expression profiles. Furthermore, there was an increase in the genomic instability after long time of in vitro expansion, and this instability was greater when cells were exposed to high doses of titanium microparticles that induced oxidative stress. It is necessary always assess the risks/ benefits of using titanium in tissue therapy involving MSCs, considering the biosafety of the use of bone regeneration using titanium and MSCs. Even without using titanium, it is important that the therapeutic use of such cells is based on analyzes that ensure quality, security and cellular stability, with the standardization of quality control programs appropriate. In conclusion, it is suggested that cytogenetic analysis, FISH analysis and the micronucleus and other nuclear alterations are carried out in CTMH before implanting in a patient

Transcriptoma diferencial entre células-tronco mesenquimais humanas jovens e senescentes

Human mesenchymal stem cells (MSC) are powerful sources for cell therapy in regenerative medicine. The long time cultivation can result in replicative senescence or can be related to the emergence of chromosomal alterations responsible for the acquisition of tumorigenesis features in vitro. In this study, for the first time, the expression profile of MSC with a paracentric chromosomal inversion (MSC/inv) was compared to normal karyotype (MSC/n) in early and late passages. Furthermore, we compared the transcriptome of each MSC in early passages with late passages. MSC used in this study were obtained from the umbilical vein of three donors, two MSC/n and one MSC/inv. After their cryopreservation, they have been expanded in vitro until reached senescence. Total RNA was extracted using the RNeasy mini kit (Qiagen) and marked with the GeneChip ® 3 IVT Express Kit (Affymetrix Inc.). Subsequently, the fragmented aRNA was hybridized on the microarranjo Affymetrix Human Genome U133 Plus 2.0 arrays (Affymetrix Inc.). The statistical analysis of differential gene expression was performed between groups MSC by the Partek Genomic Suite software, version 6.4 (Partek Inc.). Was considered statistically significant differences in expression to p-value Bonferroni correction ˂.01. Only signals with fold change ˃ 3.0 were included in the list of differentially expressed. Differences in gene expression data obtained from microarrays were confirmed by Real Time RT-PCR. For the interpretation of biological expression data were used: IPA (Ingenuity Systems) for analysis enrichment functions, the STRING 9.0 for construction of network interactions; Cytoscape 2.8 to the network visualization and analysis bottlenecks with the aid of the GraphPad Prism 5.0 software. BiNGO Cytoscape pluggin was used to access overrepresentation of Gene Ontology categories in Biological Networks. The comparison between senescent and young at each group of MSC has shown that there is a difference in the expression parttern, being higher in the senescent MSC/inv group. The results also showed difference in expression profiles between the MSC/inv versus MSC/n, being greater when they are senescent. New networks were identified for genes related to the response of two of MSC over cultivation time. Were also identified genes that can coordinate functional categories over represented at networks, such as CXCL12, SFRP1, xvi EGF, SPP1, MMP1 e THBS1. The biological interpretation of these data suggests that the population of MSC/inv has different constitutional characteristics, related to their potential for differentiation, proliferation and response to stimuli, responsible for a distinct process of replicative senescence in MSC/inv compared to MSC/n. The genes identified in this study are candidates for biomarkers of cellular senescence in MSC, but their functional relevance in this process should be evaluated in additional in vitro and/or in vivo assays

Evuloção cognitiva e funcional de pacientes pós-acidente vascular cerebral - AVC

PURPOSE: Stroke is a high-incidence cerebrovascular disease with elevated morbidity that results in impairments such as functional disabilities. This study aimed to investigate the functional evolution of individuals in the first six months post-stroke. METHOD: Longitudinal study with 42 stroke patients. The functional independence measure (FIM) and The National Institutes of Health Stroke Scale (NIHSS) were used by multidisciplinary staff 3 times in each participant; the first application was at admission to rehabilitation and the others three and six months later. RESULTS: Sample predominantly female (57%), married (52%), mean age 65.26 ±10.72 years, elementary schooling level (43%), ischemic stroke (91%), and right cerebral hemisphere (74%). Motor FIM scores and NIHSS scale showed improvement in the 3 evaluations, with significant p-value (<0.001). There was a strong relation between motor FIM evolution and NIHSS evolution (r = - 0.69 p-value< 0.001). CONCLUSIONS: It was observed that functional evolution at 6 months post-stroke was significant and the smaller the evolution of clinical impairment in these patients, the larger the evolution of their functional independence. The study is important because it allows a more appropriate therapeutic planning according with functional evolution in stroke rehabilitation

Seleção de categorias da Classificação Internacional de Funcionalidade, Incapacidade e Saúde para avaliação do paciente com acidente vascular cerebral em reabilitação

Stroke represents the first cause of disabilities among adults. Although different professions work together in treatment of stroke patients, all they use different terminologies for the description of the patients problems and it can constitute an impediment in the communication between the staff members. Thus, the multidisciplinary and interdisciplinary work would be facilitated if using a reference common tool, as the new International Classification of Functioning, Disability and Health (ICF). However, the ICF is very extensive and complex and due to its complexity, it has been evidenced the necessity to select its categories to become it more practical. The aim of the study was to investigate which categories of the ICF are more suitable to evaluate and to describe the stroke patient in the view of teachers and municipal public health professionals. It was a descriptive research, which involved 5 professors and 11 professionals of Physiotherapy that have worked at the health public area in Natal / RN. It was used the Delphi Technique in 3 rounds and the Likert Scale to select the categories among the ICF components. As result, from the 362 IFC categories, 94 were selected. The selected categories correspond to rehabilitative characteristics of Stroke patients in the universe of the Physiotherapy performance. The methodology applied was suitable to the studied object emphasizing the necessity of future studies for validation of the chosen categories

Avaliação do desempenho funcional de crianças com paralisia cerebral de acordo com níveis de comprometimento motor

Contextualization: Several studies have examined the mobility of this group of children, however little is known about the impact of motor function in activities of daily living, considering the seriousness of their neuromotor damage. Objective: Identify the functional differences of children with Cerebral Palsy with different levels of motor dysfunction and correlate these differences with the areas of mobility, self-care and social function in functional ability and caregiver´s assistance of these children. Methods: An l analytical cross-section search was developed, which were part 70 children / families aged from 4 to 7.5 years, in the Rehabilitation Center for Children. As tools were used the Pediatric Evaluation Disability Inventory (PEDI) and the Gross Motor Function Classification System (GMFCS). Data analysis was performed by ANOVA and Pearson's correlation tests. Results: The results show the functional variability of children CP in different severity levels of motor disfunction This variation was observed in the areas of mobility, self-care and social function. The results also showed a strong correlation between the domains mobility and self-care, mobility and social function. Conclusions: The variability shown by the children with CP, suggests the use of PEDI and GMFCS as this association appears to increase the understanding of how the gross motor functions are related to activities of daily living, describing the best commitments and their degree of impact on functional activities. This correlation demonstrates how mobility is crucial to evaluate the performance and guide the therapeutic practice, to develop the children´s potencial, and guide the caregiver in stimulation

Do olhar inquieto ao olhar comprometido: uma experiência de intervenção voltada para a atuação com alunos que apresentam paralisia cerebral

Compte tenu de l importance de la formation des enseignants pour répondre, de nos jours, aux besoins des élèves dont les caractéristiques marquantes sont la diversité et la différentiation, desquels nécessairement font partie , à un échelon croissant, des personnes atteintes d une déficience quelconque ; devant le constat que les actuelles politiques d éducation, en fonction de leurs limitations, peuvent être considérées comme un des principaux facteurs qui entravent la concrétisation d un vrai processus d inclusion de ces personnes ; compte tenu, encore, que le confront des situations éducatives concernant le processus d inclusion de ceux ayant des déficiences physiques, notamment lorsqu il s agit d élèves atteints de paralysie cérébrale, engendre chez les enseignants un état de troublante inquiétude du fait de ne se sentir convenablement préparés pour se confronter avec des situations pareilles, cette recherche, centrée sur cette problématique, a chosi comme objectif planifier, mettre en marche et analyser un programme d intervention pédagogique dans une école régulière de la ville de Natal/RN, auprès de quatre enseignants de l école fondamentale (7e Série) qui avaient reçu, dans leur classe, deux élèves atteints de paralysie cérébrale. S utilisant comme recours méthodologique de la recherche-action, le programme d intervention s est structuré autour de troix axes thématiques : l attitudinal, le pédagogique et le vécu en milieu scolaire, sous la forme de discussions théoriques et la mise en oeuvre de ces thématiques. Les données qui devraient être soumises à l analyse ont été collectées à partir des procédés d observation, d entretiens avant et après l intervention, du régistre de photos et d un questionnaire. Les interprétations faites, basées dans la comparaison du discours des sujets ont signalé que les enseignants ont progressé dans la maîtrise des savoirs scienfiques concernant la paralysie cérébrale et dans la connaissance des personnes atteintes par cette déficience. Par rapport aux valeurs et significations attribuées par les enseignants à ce programme d intervention à partir de leur choix des photos dans l ensemble du déroulement de ce processus formatif, elles s expriment par une prise de conscience très marquée, de la part des enseignants, par rapport :aux besoins éducatifs des élèves atteints de paralysie cérébrale ; à l importance et au vrai rôle que l école doit jouer face aux politiques d éducation inclusive ; aux difficultés vécues par ces personnes, qui réclament le plus profond respect de leurs ingularités ; la reconstruction d une nouvelle image concernant ces personnes ; la constatation qu il est possible aux enseignants d adapter et même de créer à l école des ressources et matériaux pédagogiques envisageant la qualité du processus d enseignement et apprentissage de l élève atteint de paralysie cérébrale ; finalement, la perception de l importance du travail en équipe, d un milieu scolaire accessible à ces personnes et de l appuis de la famille dans le contexte de l éducation inclusive. Les résultats obtenus, issus de ce programme d intervention, d après l évaluation finale des sujets, de part de témoigner de son efficacité, montrent que l intervention a été une excellente opportunité d habiliter ces enseignants et d améliorer le processus d enseignement à l école choisie comme le locus de cette recherche

Trilhas da inclusão escolar percorridas por uma aluna com paralisia cerebral na EJA: concepções e práticas

On this qualitative study, it has been discussed the school inclusion paths done by a student with cerebral palsy, who we call Liz, and who was also the reference and indirect subject for this study. For the representation of landscapes found, it follows throughout historical, political and pedagogical paths, getting through the case study method references on the current Brazilian educational context, analyzing, which concepts are attributed to the inclusion and which practices are developed by educational managers and teachers in a regular school of the City of Natal/RN. The theory which based this research is supported on Vygotsky's central ideas (1991, 1997, 2004) and his followers. Furthermore, it was essential to bring to this investigative journey a literature which could dialogue with the most relevant aspects of the cultural-historical approach, emphasizing the assumptions of a progressive education tendency, which promotes the subject involvement of his/her work in the world and for the world. This way, it was necessary to seek for theoretical support on assistive technology and on alternative communication, in order to show the importance of establishing other communications, which is, to break away from the conventional pattern established by the school. On this journey it was used procedures for the data construction, such as: observation, conducting semistructured interviews and questionnaires; and document analysis that supported and legitimized the inclusion, besides the permanent field diaries record. On the access paths to this landscape it entered in the Youth and Adults Education (Educação de Jovens e Adultos - EJA), seeking to establish a dialogue among Youth and Adult Education and Special Education. It was verified that, still, there are large gaps in these policies articulation. Among the results obtained it was raised discussions on new scenery, in which Youth and Adults Education (EJA) emerges as prominent mode of the established relationships throughout the school inclusion process. There were target reflections on: the planning and evaluation systematic, the pedagogical articulation among the Youth and Adults Education (EJA) teachers and the multifunctional resource room teacher's actions and the continuous training importance of the educators involved. It was considered, therefore, that the necessary mediations for school inclusion may be sustained if the walkers are involved in a permanent contact with the nature of a proposal from the Special Education inclusive perspective

Uma contribuição crítica para o redesenho de cadeiras de rodas adaptadas para crianças e adolescentes com paralisia cerebral

This work was aimed at making a critical analysis of the product wheelchair, both for using four different models, which were objects of study of the dissertation of Cláudia Regina Cabral Galvão, entitled Critical Analysis of the Mobility Products Seated a wheelchair - Used by Children and Adolescents with Cerebral Palsy in Natal / RN and other municipalities of Rio Grande do Norte . This product is considered an instrument in the social rehabilitation of great importance for people with physical disabilities. This study aims to position the issue and develop comments on technical up grading of certain models according to the needs of the user. Describes features of four models in search searched through catalogs in order to know its advantages and disadvantages of use. Were presented the definitions of ergonomics and ergonomic aspects to be considered on a design, the study of anthropometry and its recommendations. Discussions the methodology of project design in two parts: the first, on the structuring of design problem (formulation, analysis, synthesis and evaluation.) And the second on the project (design and development, implementation and evaluation and solution). With that review will include the possibilities for a new redesign of the wheelchair, based on forms of adaptation in order to achieve the target that was compressed by the average population studied. Seeks to that this project makes an improvement in quality of life of people in wheelchairs by including these people in society but also the improvement of rehabilitation

Desenvolvimento de protótipo de cadeira de banho para indivíduos com paralisia cerebral tetraparética espástica

This study proposes to develop an equipment that attends the demands of tetraplegic people due to cerebral palsy and that promotes an adequate caregivers postural biomechanics during the shower activity of daily living. First, a bibliographic review was performed to define the terms refering to cerebral palsy, activity of daily living (specifically shower), and assistive technology, besides listing the wheelchairs made on the mainly national assistive technology companies. Therefore, this is a descriptive-exploratory study based on a literature review and on a based-field exploration research. On the field research a survey was adopted as a methodological procedure as it is related to a direct investigation related to a phenomenon, on the case, represented by the current shower situation of the people investigated in this study. Data were collected with the application of a form to the caregivers and consumers of the medical-therapeutic treatment and place used by the participants. Such form, which was made up of open and close questions, tried to identify, besides the personal data of evaluated users and consumers, the characteristics of the current shower activity, such as the place where it takes place in the house, the used equipmentS, in the case there is any, and how often it occurs. The form also was used to identify the caregivers and consumers desires and perceptions in relation to the present characteristics of the new dispositive besides the users and consumers anthropometric data. The evaluation of the results obtained through the form, together with the practice and clinical experience of the researchers and engineers involved in this study, made it possible to develop and make up a real shower chair prototype with the specific adjusts destined to adequate the equipment to be used according to the needs of each user and consumer

Análise crítica dos produtos de mobilidade sentada - cadeiras de rodas - utilizados por crianças e adolescentes com paralisia cerebral em Natal/RN e outros municípios do Rio Grande do Norte

The objective of this work is to critically analyze the seated mobility adaptive equipment, wheelchair, used by non-ambulatory children/adolescents diagnosed with cerebral palsy of the quadriplegy type with ages in between 0-18 years old in the municipal city of Natal/RN and of other ones in the state of Rio Grande do Norte. This study is characterized as descriptive as it took into consideration the individualized conditions of each child/adolescent and of their home environment, and also evaluates the condition of the used seated mobility devices such as wheelchairs or strollers. Thirty three (33) subjects which had a wheelchair prescribed by a healthcare professional specialist in adaptive equipment within the past 3 years were selected. A questionnaire was applied through a phone with the patient s primary care giver or parents. The results obtained showed that all the equipment being used had an anatomical seat and back and the tilt system mechanism and even though most of the patients required small individual adjustments upon receiving the wheelchair in order to accommodate to the patients characteristics. From the point of view of the caregiver/parents interviewed, 97% described satisfied with the equipment and report assistance with mobility (97%), comfort (82%) and good posture (82%) as the main positive aspects from the equipment. The results also describe the aspects related to the maintenance, time used and activities developed that were reported by the patients. The conclusion therefore is that in order to select the equipment to meet the needs of the consumer it is necessary that: an appropriate wheelchair prescription takes place, the up-to-date knowledge of equipment available in the market, the appropriate use of the resources from the concession programs of the government that purchase equipment and the establishment of link in between the manufacturers, patients and the caregivers

Desempenho motor de pacientes com acidente vascular cerebral em um jogo baseado em realidade virtual

The Cerebral Vascular Accident (CVA) is the leading cause of motor disability in adults and elderly and that is why it still needs effective interventions that contribute to motor recovery. Objective: This study was aimed to evaluate the performance of stroke patients in chronic stage using a virtual reality game. Method: 20 patients (10 with injury to the left and 10 to the right side), right-handed, average age 50.6 ± 9.2 years, and 20 healthy subjects with average age of 50.9 ± 8.8, also right-handed participated. The patients had a motor (Fugl-Meyer) and muscle tone assessment (Ashworth). All participants made a kinematic evaluation of the drinking water activity and then underwent training with the table tennis game on XBOX 360 Kinect®, 2 sets of 10 attempts for 45 seconds, 15 minutes rest between sets, giving a total of 30 minutes session. After training the subjects underwent another kinematic evaluation. The patients trained with the right and left hemiparect upper limb and the healthy ones with the right and left upper limb. Data were analyzed by ANOVA, t Student test and Pearson correlation. Results: There was significant difference in the number of hits between the patients and healthy groups, in which patients had a lower performance in all the attempts (p = 0.008), this performance was related to a higher level of spasticity (r = - 0.44, p = 0.04) and greater motor impairment (r = 0.59, p = 0.001). After training, patients with left hemiparesis had improved shoulder and elbow angles during the activity of drinking water, approaching the pattern of motion of the left arm of healthy subjects (p < 0.05), especially when returning the glass to the table, and patients with right hemiparesis did not obtain improved pattern of movement (p > 0.05). Conclusion: The stroke patients improved their performance over the game attempts, however, only patients with left hemiparesis were able to increase the angle of the shoulder and elbow during the functional activity execution, better responding to virtual reality game, which should be taken into consideration in motor rehabilitation

Grau de conhecimento de pacientes sobre o acidente vascular cerebral e a necessidade do uso de cartilha educativa para orientação fisioterapêutica

Stroke is a neurological dysfunction of vascular origin that causes physical impairments and disabilities. Brazil leads the statistics stroke mortality among Latin American countries, demonstrating still be a neglected disease in this country. The incidence is related to risk factors and still is great misinformation in our country about stroke, treatment and prophylaxis. The aim of this study was to assess the degree of knowledge about stroke and patients need to use educational booklet for physiotherapy. The sample consisted of 53 patients, 22 females and 31 males, mean age 56.2 ± 10.9 years. Patients were evaluated by completing questionnaires, observing the socio-demographic and clinical aspects, neurological assessment, functional assessment, knowledge of pathology and presenting educational booklet. The data were analyzed using the chi-square test. According to the results, it was observed that the patients had not enough knowledge about the terminology, complications and warning signs of stroke, were not aware and did not perform the proper positioning of the limb and transfers, however, were aware and performed the stretching maneuvers. This reveals the need to implement policies that lead to information about stroke population. Following discharge from outpatient physical therapy using educational booklets with guidelines on positioning, stretching exercises and transfer is important in functional rehabilitation and encouraging independence of the individual affected by stroke

Influência da adição de carga na marcha em esteira de crianças com paralisia cerebral hemiparética espástica

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