40 resultados para tronco encefálico


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To the vertebrates, maintain body balance against the gravitational field and be able to orient themselves in the environment are fundamental aspects for survival, in which the participation of vestibular system is essential. As part of this system, the vestibular nuclear complex is the first central station that, by integrating many information (visual, proprioceptive), and the vestibular, assumes the lead role in maintaining balance. In this study, the vestibular nuclear complex was evaluated in relation to its cytoarchitecture and neurochemical content of cells and axon terminals, through the techniques of Nissl staining and immunohistochemistry for neuronal specific nuclear protein (NeuN), glutamate (Glu), substance P (SP), choline acetyltransferase (ChAT) (enzyme that synthesizes acetylcholine-Ach) and glutamic acid decarboxylase (GAD) (enzyme that synthesizes gamma-amino butyric acid-GABA). The common marmoset (Callithrix jacchus) was used as experimental animal, which is a small primate native from the Atlantic Forest in the Brazilian Northeast. As results, the Nissl technique, complemented by immunohistochemistry for NeuN allowed to delineate the vestibular nucleus superior, lateral, medial and inferior (or descending) in the brain of the common marmoset. Neurons and terminals immunoreactive to Glu and ChAT and only immunoreactive terminals to SP and GAD were seen in all nuclei, although in varying density. This study confirms the presence in the vestibular nuclei of the common marmoset, of Glu and SP in terminals, probably from the first order neurons of vestibular ganglion, and of GABA in terminals, presumably from Purkinge cells of the cerebellum. Second-order neurons of the vestibular nuclei seem to use Glu and Ach as neurotransmitters, judging by their expressive presence in the cell bodies of these nuclei in common marmosets, as reported in other species

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The parabrachial complex (PB) is an area of the brainstem responsible for the processing and transmission of essential physiologic information for the survival of the organisms. This region is subdivided in approximately nine subregions, considering morphology, cytoarchitectural and functional characteristic. Its neurons have an extensive network of connections with other regions of the nervous system. The objective in this work was to map the retinal projection to the PB and make a citoarchitectonic and neurochemical characterization of this region in the common marmoset (Callithrix jacchus), a primate of the New World. The retinal projections were mapped by anterograde transport of the choleric toxin subunit b (CTb). The citoarchitecture was described through the Nissl method, and the neurochemical characterization was made through immunohistochemical technique to the some neurotransmitters and neuroactives substances present in this neural center. In marmoset PB, in the coronal sections labeled by Nissl method, we found a similar pattern to that evidenced in other animal species. The immunoreactivity against CTb was verified in the PBMv in fibers/terminal, characterizing such as retinal innervations in this area. The immunohistochemical technique reveled that the PB contain cells, fibers and/or terminals immunoreactives to the neuronal nuclear protein, Choline acetyl transferase, nitric oxide synthase, serotonin, enkephalin, substance P, Calcium-binding proteins (calbindin, calretinin e parvalbumin), and glial fibrillary acidic protein. The histochemical technique reveled cells and fibers NADPH-diaphorase reactive. Each one of those substances presented a characteristic pattern of demarcation in PB, and some serve as specific markers of subregions

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Serotonin or 5-hydroxytryptamine (5-HT) is a substance found in many tissues of the body, including as a neurotransmitter in the nervous system, in which may exert varied post-synaptic actions. Inside the neuro-axis, the location of 5-HT neurons is almost restricted to the raphe nuclei of the brainstem, such that 5-HT-immunoreactivity can be considered a marker of the raphe nuclei. The raphe nuclei are located in the brainstem, at or near the midline. The serotonergic groups were originally alphanumerically classified as B1 to B9 towards caudorrostral in rats and can be divided into upper and lower groups. In this study the distribution of serotonergic neurons was studied using immunohistochemistry in the brain of the rock cavy (Kerodon rupestris), a species of rodent endemic to Northeastern Brazil. The cytoarchitectonic location of serotonergic neurons was established in series of adjacent coronal and sagittal sections stained by the Nissl method and immunohistochemistry for 5-HT. Thus, we defined the raphe rostral linear, caudal linear, dorsal, median, and paramedian pontine raphe nuclei, and B9 cluster, constituting the rostral group, and the interpositus, magnus, obscure and palidus, constituting the caudal part of the group, comparable to which has been described for other mammalian species

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The hypothalamus is a diencephalic portion located around the third ventricle below the hypothalamic sulcus, limited by the optic chiasm, and by the mammillary bodies, acting as a center that integrates behavioral and homeostatic functions. Serotonin is a neurotransmitter produced in limited sites in the midbrain and brain stem, but is distributed throughout the central nervous system and has many functions, acting through specific receptors that are also distributed throughout the nervous system. Using immunohistochemical techniques, the aim of this study was to delineate the hypothalamic nuclei of the marmoset (Callithrix jacchus) and study the distribution of serotonin transporter and serotonin receptors in the hypothalamus of this species. We used the Nissl method to determine the cytoarchitecture of the hypothalamic nuclei, and immunohistochemistry to reveal the presence of NeuN as a method to determine the contours of the hypothalamic nuclei. As a result, we found serotonin containing fibers and terminals throughout the rostrocaudal extent of the hypothalamus, more concentrated in some nuclei, and even absent in some. Like serotonin, serotonin transporter was observed between pre-optic area and tuberal region of the hypothalamus, in densities and distribution similar to serotonin. The 5-HT1A and 5-HT1B receptors were found with minor differences among itselves regarding the disposition and intensity of staining.

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The midline/intralaminar nuclei form a remarkable group of nuclei of the medial and dorsal thalamus. The midline nuclei, in rats, comprises the paratenial nuclei (PT), paraventricular (PV), intermediodorsal (IMD), reuniens (Re) and rhomboid (Rh). The intralaminar nuclei comprises the central medial (CM), paracentral (PC), central lateral (CL) and parafascicular (PF). Such nuclei have dense serotonergic innervation originating from the brainstem, especially from the so-called ascending activation system. These nuclei, in turn, send projections to various cortical and subcortical areas, specifically to limbic areas, which suggests the important role of this neurotransmitter in the limbic circuitry. The aim of this study was to characterize the distribution pattern and morphology of serotonin fibers in the nuclei of the midline and intralaminar thalamic of rocky cavy (Kerodon rupestris), a tipical rodent from brazilizan northeast. To reach this aim we used four rock cavies adults. Following the transcardially perfusion with paraformaldehyde and brain microtomy steps was performed immunohistochemistry for serotonin (5-HT), Nissl technique and subsequent achievement and image analysis to characterize the cytoarchitecture of these nuclei and the serotonergic fibers visualized. An analysis was made of Relative Optical Density (ROD) to semi-quantify the concentration of serotonin fibers in the areas of interest. Thus, we observed a cytoarchitectonic arrangement of these nuclei similar to that found in rats. In case of fibers distribution, those immunoreactive to 5-HT were presented in a higher concentration according as ROD in the midline nuclei relative to intralaminar; Re being the core which has a higher pixel value followed by the PV , Rh, IMD and PT. In intralaminar CL showed higher pixels, followed by nuclei CM, PC and PF. The serotonergic fibers were classified as number of varicosities and axon diameter, therefore find three types of fibers distributed through this nuclear complex: fibers rugous, granular and semi-granular. In PV fibers predominated rugous; in PT fibers predominated granular; IMD, CL and PF fibers were represented by semi-granular and Re, Rh, PC and CM fibers showed granular and semi-granular. Morphological characterization of serotonergic fibers and differences in density between the nuclei may suggest different patterns of synaptic organization of this neurotransmitter beyond confirming his large repertoire functional

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Serotonin or 5-hydroxytryptamine (5-HT) is a substance found in many tissues of the body, including the nervous system acting as a neurotransmitter. Within the neuro-axis, the location of the majority of the 5-HT neurons is superimposed with raphe nuclei of the brain stem, in the median line or its vicinity, so that neuronal 5-HT can be considered a marker of the raphe nuclei. Serotonergic neurons are projected to almost all areas of the brain. Studies show the participation of serotonin in regulating the temperature, feeding behavior, sexual behavior, biological rhythms, sleep, locomotor function, learning, among others. The anatomy of these groups has been revised in many species, including mouse, rabbit, cat and primates, but never before in a bat species from South America. This study aimed to characterize the serotonergic clusters in the brain of the bat Artibeus planirostris through immunohistochemistry for serotonin. Seven adult bat males of Artibeus planirostris species (Microchiroptera, Mammalia) were used in this study. The animals were anesthetized, transcardially perfused and their brains were removed. Coronal sections of the frozen brain of bats were obtained in sliding microtome and subjected to immunohistochemistry for 5-HT. Delimit the caudal linear (CLi), dorsal (DR), median (MnR), paramedian (PMnR), pontine (PNR), magnus (MgR), pallidus (RPA) and obscurus (ROb) raphe nucleus, in addition to the groups B9 and rostral and caudal ventrolateral (RVL/CVL). The serotonergic groups of this kind of cheiroptera present morphology and cytoarchitecture relatively similar to that described in rodents and primates, confirming the phylogenetic stability of these cell clusters.

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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

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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

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It is known that sleep plays an important role in the process of motor learning. Recent studies have shown that the presence of sleep between training a motor task and retention test promotes a learning task so than the presence of only awake between training and testing. These findings also have been reported in stroke patients, however, there are few studies that investigate the results of this relationship on the functionality itself in this population. The objective of this study was to evaluate the relationship between functionality and sleep in patients in the chronic stage of stroke. A cross-sectional observational study was conducted. The sample was composed of 30 stroke individuals in chronic phase, between 6 and 60 months after injury and aged between 55 and 75 years. The volunteers were initially evaluated for clinical data of disease and personal history, severity of stroke, through the National Institute of Health Stroke Scale, and mental status, the Mini-Mental State Examination. Sleep assessment tools were Pittsburgh Sleep Quality Index, the Questionnaire of Horne and Ostberg, Epworth Sleepiness Scale, the Berlin questionnaire and actigraphy, which measures were: real time of sleep, waking after sleep onset, percentage of waking after sleep onset, sleep efficiency, sleep latency, sleep fragmentation index, mean activity score. Other actigraphy measures were intraday variability, stability interdiária, a 5-hour period with minimum level of activity (L5) and 10-hour period with maximum activity (M10), obtained to evaluate the activity-rest rhythm. The Functional Independence Measure (FIM) and the Berg Balance Scale (BBS) were the instruments used to evaluate the functional status of participants. The Spearman correlation coefficient and comparison tests (Student's t and Mann-Whitney) were used to analyze the relationship of sleep assessment tools and rest-activity rhythm to measures of functional assessment. The SPSS 16.0 was used for analysis, adopting a significance level of 5%. The main results observed were a negative correlation between sleepiness and balance and a negative correlation between the level of activity (M10) and sleep fragmentation. No measurement of sleep or rhythm was associated with functional independence measure. These findings suggest that there may be an association between sleepiness and xii balance in patients in the chronic stage of stroke, and that obtaining a higher level of activity may be associated with a better sleep pattern and rhythm more stable and less fragmented. Future studies should evaluate the cause-effect relationship between these parameters

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The cerebral vascular accident is a neurological dysfunction of vascular origin that leds to development of motor sensibility, cognitive, perceptive and language deficits. Despite the fact that the main sleep disorders in stroke patients are well known, it is still necessary to analyze which mechanisms of regulation of sleep and wakefulness are affected. The objective of this study was to evaluate the changes in the circadian and homeostatic control of sleep-wakefulness in stroke patients and the correlations with quality of life and level of physical activity. The study analyzed 22 stroke patients (55± 12 years old) and 24 healthy subjects (57 ±11 years old). The instruments used in this study were questionnaires on sleep quality, daytime sleepiness, quality of life, physical activity level and the actigraphy. The data were analyzed using the Student `t test, Mann-Whitney test, ANOVA and Spearman's correlation tests. The results showed stability in the sleep-wake circadian expression with changes in the amplitude of the rhythm. However, significant changes were found related to the homeostatic component characterized by increased sleep duration, increased latency, fragmented sleep and lower sleep efficiency. Additional data showed decreased quality of sleep and increased daytime sleepiness, as well as decreased quality of life and level of physical activity. The results indicate that the interaction of circadian and homeostatic control of sleep-wake is compromised and the main reason might be because of the homeostatic component and the lower activity level resulting from the brain damage. Thus, further studies may be developed to evaluate whether behavioral interventions such as increased daytime activity and restriction of sleep during the day can influence the homeostatic process and its relation to circadian component, resulting in improved quality of nocturnal sleep in stroke patients

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observar os efeitos agudos de diferentes intensidades de Pressão Expiratória Positiva (PEP) sobre a cinemática do complexo toracoabdominal de pacientes acometidos por Acidente Vascular Encefálico (AVE). Métodos: Foram selecionados 21 indivíduos com AVE e 16 indivíduos saudáveis pareados por idade sexo e IMC para grupo controle. Avaliamos função pulmonar, pressões dos músculos respiratórios e os volumes pulmonares por meio da Pletismografia Optoeletrônica durante três diferentes intensidades de PEP 10, 15 e 20 cmH2O. Resultados: o efeito da PEP no volume corrente (VC) do grupo AVE em relação ao grupo controle foi diferente. Enquanto o grupo controle aumentou o VC em relação a respiração tranquila em 343%, 395,2% e 431,8% nas PEP10, PEP15 e PEP20 cmH2O o grupo AVE aumento 186%, 218.8% e 209.5% (p < 0.0001). A PEP também influenciou de forma diferente em relação ao Tempo inspiratório com intensidades diferentes no grupo controle e AVE (p < 0.0001). No ciclo de trabalho foi observado um aumento no grupo controle nas PEP10 (p < 0.001) e PEP15 (p < 0.05) e no grupo AVE foi observada uma redução PEP20 (p < 0.01) quando comparada com a respiração tranquila. Os volumes operacionais do grupo AVE foi observado aumento do volume inspiratório final da parede torácica (Vifpt) e do Volume expiratório final da parede torácica (Vefpt) diferente do grupo controle que gerou aumento do Vifpt acompanhado de diminuição do Vefpt durante as três intensidades de PEP. Conclusão: A hiperinsuflação observada no grupo AVE demonstra que essa terapêutica deve ser utilizada com cautela especialmente nas intensidades maiores que 10 cmH2O para essa população

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Mesenchymal stem cells (MSCs) are known as a population of multi-potential cells able to proliferate and differentiate into multiple mesodermal tissues including bone, cartilage, muscle, ligament, tendon, fat and stroma. Several applications of the study of EC can be emphasized the therapeutic techniques such as guided bone regeneration by implantation of EC in the affected site, without the need for bone grafts, using titanium as a vehicle. The process of cryopreservation is essential for the maintenance of cell cultures, since the cell line is frozen, it can be maintained in liquid nitrogen for an indefinite period and then thawed for therapeutic or experimental purposes. The aim of this study was to isolate a population of MSCs derived from the subendothelium of the umbilical vein human (MSCs-SUVH) to assess cytogenetic analysis by the possibility of appearance of chromosomal changes in two different situations: MSCs-SUVH regarding the process of cryopreservation and MSCs-SUVH grown on the surface of titanium. Flow cytometry analysis revealed that, this cell population was positive for the markers CD29, CD73 and CD90, but there was no expression of hematopoietic lineage markers, such as CD14, CD34 and CD45 and demonstrated capacity for osteogenic differentiation. The chromosomes obtained from the primary culture of MSCs-SUVH were analyzed by GTW banding technique, and results are described as guidelines to ISCN 2005. There was not the emergence of clonal chromosomal changes in the MSCs-SUVH in different situations analyzed. However one of the strings presented a balanced paracentric inversion, probably a cytogenetic constitutional alterations, which was present before and after the experimental situations that the MSCs-SUVH was submitted

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A number of evidences show the influence of the growth of injured nerve fibers in Peripheral Nervous System (PNS) as well as potential implant stem cells (SCs) to make it more suitable for nerve regeneration medium. In this perspective, this study aimed to evaluate the plasticity of mesenchymal stem cells from bone marrow of mice in the presence of culture medium conditioned with facial nerve explants (D-10) and fibroblast growth factor-2 (FGF-2). In this perspective, the cells were cultivated only with DMEM (group 1), only with D-10(group 2), only with FGF-2(group 3) or with D-10 and FGF-2(group 4). The growth and morphology were assessed over 72 hours. Quantitative phenotypic analysis was taken from the immunocytochemistry for GFAP, OX-42, MAP-2, β-tubulin III, NeuN and NF-200 on the fourth day of cultivation. Cells cultured with conditioned medium alone or combined with FGF-2 showed distinct morphological features similar apparent at certain times with neurons and glial cells and a significant proliferative activity in groups 2 and 4 throughout the days. Cells cultived only with conditioned medium acquired a glial phenotype. Cells cultured with FGF-2 and conditioned medium expressed GFAP, OX-42, MAP-2, β-tubulin III, NeuN and NF-200. On average, area and perimeter fo the group of cells positive for GFAP and the área of the cells immunostained for OX-42 were higher than those of the group 4. This study enabled the plasticity of mesenchymal cells (MCs) in neuronal and glial nineage and opened prospects for the search with cell therapy and transdifferentiation

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During the motor rehabilitation of patients with neurological pathologies, there are many factors that have an influence on the therapeutical process. It is possible that the demonstration using videotapes or photographs of movements that will be learned can assist the codification, classification and reorganization of the elements of the task in familiar diagrams and thus facilitate the process of motor rehabilitation. The aim of this study was to analyze the performance in the test of recognition of sequences of functional activities such as water drinking and hair combing presented partially (by photos) and completely (by video) to the patients with encephalic vascular accident . The sample was generated from 12 patients (62 ± 8 years) and 10 healthy subjects (53 ± 5 years), of both sex, divided in two groups for which 5 functional activities were presented partially and completely, in 4 sequences that varied between reached and not reached objectives and correct and incorrect movements, during 24 trials. The test t'Student was applied to verify differences in the recognition time between the groups, and to compare the performance between the tests carried through at 9:00 am and retests at 9:00 am and 4:00 pm. To compare the frequency between the types of reply, the test Qui-square was used. It was verified that the patients presented a greater recognition time and a lower number of correct answers than the healthy subjects, and had better performances in the recognition of the complete sequence than the partial one. Bigger frequency was observed in the reply that the objective and the movement were being presented correctly, however the healthy subjects had soon indicated the correct reply in the first trials. An improvement tendency was observed in the performance, when the stages of tests and retests had been carried through in the same schedule. According to the results, it is suggested that the patients with stroke can better recognize the correct movements of a functional activity when it is presented completely in video form, what can influence the choice of the therapeutical strategy during the process of motor rehabilitation

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During the motor rehabilitation of patients with neurological pathologies, there are many factors that have an influence on the therapeutical process. It is possible that the demonstration using videotapes or photographs of movements that will be learned can assist the codification, classification and reorganization of the elements of the task in familiar diagrams and thus facilitate the process of motor rehabilitation. The aim of this study was to analyze the performance in the test of recognition of sequences of functional activities such as water drinking and hair combing presented partially (by photos) and completely (by video) to the patients with encephalic vascular accident . The sample was generated from 12 patients (62 ± 8 years) and 10 healthy subjects (53 ± 5 years), of both sex, divided in two groups for which 5 functional activities were presented partially and completely, in 4 sequences that varied between reached and not reached objectives and correct and incorrect movements, during 24 trials. The test t'Student was applied to verify differences in the recognition time between the groups, and to compare the performance between the tests carried through at 9:00 am and retests at 9:00 am and 4:00 pm. To compare the frequency between the types of reply, the test Qui-square was used. It was verified that the patients presented a greater recognition time and a lower number of correct answers than the healthy subjects, and had better performances in the recognition of the complete sequence than the partial one. Bigger frequency was observed in the reply that the objective and the movement were being presented correctly, however the healthy subjects had soon indicated the correct reply in the first trials. An improvement tendency was observed in the performance, when the stages of tests and retests had been carried through in the same schedule. According to the results, it is suggested that the patients with stroke can better recognize the correct movements of a functional activity when it is presented completely in video form, what can influence the choice of the therapeutical strategy during the process of motor rehabilitation