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

Construção, caracterização e aplicação de eletrodos para descarga por arco de cátodo oco

The system built to characterize electrodes and, consequently, deposited fine films are constituted by a hollow cathode that works to discharges and low pressures (approximately 10-3 to 5 mbar), a source DC (0 to 1200 V), a cylindrical camera of closed borossilicato for flanges of stainless steel with an association of vacuum bombs mechanical and spread. In the upper flange it is connected the system of hollow cathode, which possesses an entrance of gas and two entrances for its refrigeration, the same is electrically isolated of the rest of the equipment and it is polarized negatively. In front of the system of hollow cathode there is a movable sample in stainless steel with possibility of moving in the horizontal and vertical. In the vertical, the sample can vary its distance between 0 and 70 mm and, in the horizontal, can leave completely from the front of the hollow cathode. The sample and also the cathode hollow are equipped with cromel-alumel termopares with simultaneous reading of the temperatures during the time of treatment. In this work copper electrodes, bronze, titanium, iron, stainless steel, powder of titanium, powder of titanium and silício, glass and ceramic were used. The electrodes were investigated relating their geometry change and behavior of the plasma of the cavity of hollow cathode and channel of the gas. As the cavity of hollow cathode, the analyzed aspects were the diameter and depth. With the channel of the gas, we verified the diameter. In the two situations, we investigated parameters as flow of the gas, pressure, current and applied tension in the electrode, temperature, loss of mass of the electrode with relationship at the time of use. The flow of gas investigated in the electrodes it was fastened in a work strip from 15 to 6 sccm, the constant pressure of work was among 2.7 to 8 x 10-2 mbar. The applied current was among a strip of work from 0,8 to 0,4 A, and their respective tensions were in a strip from 400 to 220 V. Fixing the value of the current, it was possible to lift the curve of the behavior of the tension with the time of use. That curves esteem in that time of use of the electrode to its efficiency is maximum. The temperatures of the electrodes were in the dependence of that curves showing a maximum temperature when the tension was maximum, yet the measured temperatures in the samples showed to be sensitive the variation of the temperature in the electrodes. An accompaniment of the loss of mass of the electrode relating to its time of use showed that the electrodes that appeared the spherical cavities lost more mass in comparison with the electrodes in that didn't appear. That phenomenon is only seen for pressures of 10-2 mbar, in these conditions a plasma column is formed inside of the channel of the gas and in certain points it is concentrated in form of spheres. Those spherical cavities develop inside of the channel of the gas spreading during the whole extension of the channel of the gas. The used electrodes were cut after they could not be more used, however among those electrodes, films that were deposited in alternate times and the electrodes that were used to deposit films in same times, those films were deposited in the glass substrata, alumina, stainless steel 420, stainless steel 316, silício and steel M2. As the eletros used to deposit films in alternate time as the ones that they were used to deposit in same times, the behavior of the thickness of the film obeyed the curve of the tension with relationship the time of use of the electrode, that is, when the tension was maximum, the thickness of the film was also maximum and when the tension was minimum, the thickness was minimum and in the case where the value of the tension was constant, the thickness of the film tends to be constant. The fine films that were produced they had applications with nano stick, bio-compatibility, cellular growth, inhibition of bacterias, cut tool, metallic leagues, brasagem, pineapple fiber and ornamental. In those films it was investigated the thickness, the adherence and the uniformity characterized by sweeping electronic microscopy. Another technique developed to assist the production and characterization of the films produced in that work was the caloteste. It uses a sphere and abrasive to mark the sample with a cap impression, with that cap form it is possible to calculate the thickness of the film. Through the time of life of the cathode, it was possible to evaluate the rate of waste of its material for the different work conditions. Values of waste rate up to 3,2 x 10-6 g/s were verified. For a distance of the substratum of 11 mm, the deposited film was limited to a circular area of 22 mm diameter mm for high pressures and a circular area of 75 mm for pressure strip. The obtained films presented thickness around 2,1 µm, showing that the discharge of arch of hollow cathode in argon obeys a curve characteristic of the tension with the time of life of the eletrodo. The deposition rate obtained in this system it is of approximately 0,18 µm/min

Fissura placentária de gatas SRD, Felis catus - Linnaeus, 1758: aspectos macro e microscópicos

Nossa pesquisa consiste no estudo esquemático macroscópico na placenta de gatos e a sua caracterização como tipo, placenta zonária, que 62,5% dos casos apresenta uma fissura na área distal do funículo umbilical. Esse é formado por uma área justa fetal, área justa placentária e área média, encontrando achados histológicos de 2 artérias, uma veia, 2 pedículos vitelínicos e 2 pedículos alantoidianos. Na fissura, encontramos um epitélio alantoidiano cobrindo esta área em 10% dos casos e, em 90% dos achados foram encontrados um trofoblasto diminuído comparado com outras áreas placentárias fora da fissura. Portanto, a placenta felina, com sua relação materno fetal mostra uma placenta zonária incompleta, diferente do ocorrido nos outros carnívoros.

Corpo e Velhice: um estudo da qualidade de vida de idosos da Vila Vicentina Júlia Freire em João Pessoa

The aim of this study was to follow the daily routine of elderly individuals residing at the Vila Vicentina Júlia Freire (VVJF) nursing home in João Pessoa, Brazil, observing and analyzing as much as possible, the way in which they live in a group and how they deal, through personal experiences, with their own body image and with that of others. The results were obtained from information considered relevant, according to the perspectives of the residents themselves, contained in narratives of their history and related to their quality of life. Our interest lays in how they coped with changes in their bodies at old age and adapted to the challenges of this new phase. To obtain this information, with significant expressions, we used the Narrative Interview (NI) as a research tool in order to redeem their life experience and develop an understanding of their impressions about the life experience at the current nursing home. We selected, among the housed, ten subjects - actors - to appear as a sample of the group together. The design of our sample had a hybrid model, combining two types: random sample (probability) and an intentional sample (not probability). After recording transcripts and ensuring that all the subjects showed common interests that were relevant to the group, we chose three categories for analysis as follows: a. the elderly as seen by themselves and by others; b. religiosity as a moral factor; c. intimacy and relationships. Our choice was based on the importance of the theme as a multidisciplinary topic in the quality of life of populations. The subjects, chosen as spokespersons for the VVJF, spoke, among other things, about the way in which they viewed the world, their own body and how they experienced the environment and their fellow residents, in an attitude of reflection and criticism that confirms the lucidity of an old age conditioned to an elderly institution. The method allowed us to investigate our subjects from a multidisciplinary point of view. We suggest other studies in assorted areas of the human sciences, having as principle, man at the center of all our scientific questionings. The paradigm that the elder is, necessarily, unhappy in his sheltered has been, in this job, broken through the testimonies collected, establishing multiple perspectives of the analyzed subjects by their respective narrative records. Involving the participation of researchers with major in Administration, Communication and Law, this study filled, in this aspect, the requirements of the multidisciplinarity of the Program of Pos-Graduation in Health Science

Efeito antitrombótico de uma fucana não anticoagulante extraída da alga Spatoglossum Schröederi

Fucan is a term used to denominate a family of sulfated L-fucose-rich polysaccharides. The brown alga Spatoglossum schröederi (Dictyotaceae) has three heterofucans namely fucan A, B and C. The 21 kDa fucan A is composed of a core of β (1-3) glucuronic acid-containing oligosaccharide of 4.5 kDa with branches at C4 of fucose chains α (1-3) linked. The fucose is mostly substituted at C4 with a sulfate group and at C2 with chains of β (1-4) xylose. This fucan has neither anticoagulant (from from 0.1 to 100µg) nor hemorrhagic activities (from 50 to 800 µg/mL). The antithrombotic test in vivo showed the fucan A has no activity in any of the concentrations (from 0.2 to 20µg/g/day) tested 1h after polysaccharide administration. However, when fucan A was injected endovenously 24h before the ligature of the venae cavae, we observed a dose-dependent effect, reaching saturation at around 20g/g of rat weight. In addition, this effect is also time-dependent, reaching saturation around 16h after fucan administration. In addition, regardless of administration pathway, fucan A displayed antithrombotic action. The exception was the oral pathway. Of particular importance was the finding that fucan A stimulates the synthesis of an antithrombotic heparan sulfate from endothelial cells like heparin. The hypothesis has been raised that in vivo antithrombotic activity of fucan A is related to the increased production this heparan. Taken together with the fact that the compound is practically devoid of anticoagulant and hemorrhagic activity suggests that it may be an ideal antithrombotic agent in vivo

Avaliação da terapia comunitária na atenção primária à saúde

A busca por um modelo democrático de saúde despertou a atenção do governo brasileiro para o estabelecimento de prioridades e estratégias, que impulsionaram a implantação do Programa de Saúde da Família (PSF), atualmente denominada Estratégia Saúde da Família (ESF), a fim de aproximar a equipe de saúde da comunidade e, assim, implementar ações de promoção da saúde e de prevenção do adoecimento. Nessa perspectiva a Terapia Comunitária (TC) emerge como uma tecnologia de cuidado voltada à saúde mental na Atenção Básica de Saúde. Desde 2007, a TC vem sendo desenvolvida no município de João Pessoa/PB por profissionais da ESF: enfermeiras, agentes comunitários de saúde, médicos, odontólogos, fisioterapeutas, nutricionistas, psicólogos, entre outros. O estudo teve como objetivos: avaliar a satisfação dos usuários em relação à TC na Atenção Básica no município de João Pessoa/PB; medir o nível de satisfação dos participantes da TC em relação a essa ferramenta do cuidado; identificar elementos importantes para a satisfação em relação à TC por parte dos usuários. Trata-se de um estudo avaliativo, transversal e observacional, realizado no período de maio a agosto de 2009. Utilizou-se como instrumento de coleta de dados a Escala de Avaliação da Satisfação dos Usuários com os Serviços de Saúde Mental Satis-BR, bem como um instrumento de perguntas complementares utilizado pelos terapeutas comunitários. Os resultados revelaram que dos 198 (100%) entrevistados, 105 (53%) verbalizaram satisfação e 93 (47%) muita satisfação nos encontros de TC, o que evidencia que a totalidade da amostra está satisfeita com a terapia. Os elementos importantes que concorreram para a satisfação dos usuários da TC foram: respeito, dignidade, escuta, compreensão, acolhimento, apoio nas necessidades e boas instalações dos locais onde ocorre a terapia. A TC vem fortalecendo o cuidado à saúde mental, por se constituir como uma tecnologia de prevenção e fortalecendo a porta de entrada para a rede de saúde mental e de apoio psicossocial. Conclui-se, portanto, que a TC vem se destacando como instrumento de inclusão da saúde mental na Atenção Básica no atendimento aos usuários do Sistema Único de Saúde

Avaliação do cuidado na estratégia saúde da família em metrópoles do nordeste brasileiro

Objective: To evaluate the implementation of the Family Health Strategy (FHS) in Brazilian cities of the Northeast, expanding coverage, analyzing the progress, challenges and innovations. Methods: Multicentric Evaluation Research, Studies Baselines in urban centers, using as a case study method. Selected cases of Aracaju, being capital, advanced coverage with extended team, and Fortaleza, capital coverage incipient and minimal staff. In Fortaleza, purposive sample of 11 Units Primary (APS), 03 managers, 53 professionals and 109 users. In Aracaju, 09 units of APS, 02 managers, 36 professionals, and 90 users. Structured interviews for managers, and structured to professionals and users. Descriptive analysis focusing on the political and institutional dimensions, organization and comprehensive care. Results: There was consensus that the ESF is the preferred port users and acts as inducing changes in care. In the case of Fortaleza, the specificities were: care protocols and community activities aimed at chronic conditions (100%) , with greater participation of doctors and nurses (93%) ; conjunction with more complex services, but the teams reported difficulties with the examination center and experts, the long waits and poor access to local services were the main difficulties reported by users., As innovative practice, the therapeutic group of elderly caregivers mentioned by respondents; There was intersectoral initiatives and teams 87 % of users have participated in meetings about health problems. In the case of Aracaju, care protocols were directed to the lines of care and formulated locally, 85 % coverage of the population with FHS counterpart local financing; employees hired by public tender; 70 % of teams with expertise in public health center for continuing education acting; democratization in management; access technologies, welcoming and computerization in different integrated networks, and evaluation matrix. Conclusions: The ESF has promoted access to health care and inclusion of disadvantaged populations. Different perceptions and practices in the organization of care, with distinct trajectories of reorganization. In the case of Fortaleza, predominance of model programs valuing older, with evidence of advances in care practices and teamwork, but restricted to primary care practices and incipient in public policy perspective. In Aracaju, had network integration with technologies related to the family, in which the ESF is consolidated as public policy. It can be argued that the XII APS expanding coverage, exhibited efficacy, despite the challenges inherent to the different degrees of implementation

Enxertia em citros por substituição de ápice caulinar

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Pegamento e crescimento inicial de enxertos do pessegueiro 'Aurora-1' em clones de umezeiro (Prunus mume Sieb. et Zucc.) e 'Okinawa' [Prunus persica (L.) Batsch] propagados por estacas herbáceas

O presente estudo teve por objetivo avaliar o pegamento e o crescimento inicial de enxertos do pessegueiro 'Aurora-1' em clones de umezeiro (Prunus mume Sieb. et Zucc.) e 'Okinawa' [Prunus persica (L.) Batsch] propagados por estacas herbáceas. Realizaram-se dois experimentos, adotando-se a enxertia de borbulhia por escudo (março) e borbulhia por escudo modificada (julho). Com os resultados obtidos, pode-se concluir que é viável a realização da enxertia do 'Aurora-1' nos Clones 05; 10 e 15 de umezeiro e no 'Okinawa', tanto em março quanto em julho, com as metodologias utilizadas. O 'Okinawa' induz crescimento mais rápido ao enxerto, de forma que o ponto máximo do comprimento é atingido em tempo menor.

Distribuição do sistema radicular do pessegueiro 'Okinawa' propagado por sementes e por estacas herbáceas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Entre parentes :cotidiano, religiosidade e identidade na Serra de Portalegre/RN

Si les études sur les groupes noirs ruraux connaissent un essor dans l´Antropologie brésilienne contemporaine, au Nordeste, et spécialement au Rio Grande do Norte, ce genre de recherches restent rares. Ce travail a comme principal objectif de faire une description ethnographique des communautés du Pêga, do Arrojado e do Engenho Novo, dans la commune de Portalegre/RN, désignées comme des descendants africains et dont les membres se reconnaissent comme appartenant a une "grande famille". Ainsi, nous voulons analyser les processus d´élaboration d´une identité collective et comprendre le rôle des relations de parenté dans la vie cotidienne, festive et religieuse de Portalegre. En effet, nous pouvons observer que les "parents" développent des stratégies économiques et affirment leur alterité, reprennant des traits culturels et religieux qui leur sont propres pour se présenter comme un groupe doté d´une grande cohésion face a la société environnante. Nous utilisons la méthode ethnographique et la technique de l´observation participante, alliée à l´analyse des sources historiques primaires dans le but de décrire ces pratiques et stratégies. Nous pensons que la participation des membres du groupe à la production agricole, spécialement dans la confection de la farine de manioc, la construction des relations sociales au quotidien et l´organisation de la danse de São Gonçalo représentent autant d´occasions où la mémoire du groupe se fortifie, l´identité collective se cristallise et où la sociabilité se trouve consolidée. Nous analysons ces pratiques collectives et ces moments festifs comme des moyens de réaffirmation d´une identité locale centrée sur les relations de parenté

Técnica para avaliação da pressão arterial pulmonar de equinos durante o exercício progressivo em esteira rolante

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bispectral index in dogs with high intracranial pressure, anesthetized with propofol and submitted to two levels of FiO2

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Declínio dos citros: algo a ver com o sistema de produção de mudas cítricas?

Na atualidade, a citricultura paulista enfrenta sérios problemas com o surgimento de novas enfermidades limitantes ao processo produtivo. Uma das alternativas para prevenir essas enfermidades é, seguramente, a produção de mudas certificadas, com borbulhas e sementes de assegurada qualidade genética e sanitária, produzidas em containers ou sacos plásticos, sob fiscalização dos órgãos competentes, garantindo-se, assim, a sanidade das mesmas. No entanto, tem-se verificado que grande parte do processo operacional de produção de mudas cítricas foi parcialmente adaptado da produção de essências florestais (eucalipto), onde se insere, principalmente, o emprego de tubetes com comprimento de 12 cm. Tal prática desencadeia uma grave deformidade morfológica no sistema radical das mudas cítricas, reduzindo o seu potencial de crescimento, quando são plantadas a campo, em local definitivo. Como a principal causa desencadeadora do declínio dos citros parece ser o estresse hídrico, plantas oriundas desse sistema de produção de mudas, mostram-se muito mais vulneráveis ao estresse e, conseqüentemente, a esta anomalia. Como atualmente todas as mudas produzidas no Estado de São Paulo provêm desse sistema de produção, seguramente o declínio dos citros tenderá a ser, futuramente, muito mais freqüente e severo nos casos de combinações vulneráveis à sua ocorrência.