Vascular casts of three chondrichthian, one dipnoan, one chondrostean and 14 teleostean species were examined by light and scanning electron microscopy in order to give a qualitative and quantitative analysis of interarterial anastomoses (iaas) that indicate the presence (or absence) of a secondary vascular system (SVS). Anastomoses were found to originate from a variety of different primary blood vessels, many of which have not been previously identified as giving rise to secondary vessels. Segmental arteries derived from the dorsal aorta and supplying body musculature were major sites of origin of the SVS, although there was considerable variation in where, in the hierarchy of arterial branching, the anastomoses occurred. The degree of investment in a SVS was species specific, with more active species having a higher degree of secondary vascularisation. This difference was quantified using an absolute count of iaas between Anguilla reinhardtii and Trachinotus baillonii. A range of general features of the SVS is also described. No evidence of iaas was found on the coeliac, mesenteric or renal circulation in any species. Evidence of interarterial anastomoses were lacking in the dipnoan (Sarcopterygii) and chondrichthyan species examined, suggesting that a SVS is restricted to actinopterygian fishes. The presence and distribution of a secondary vascular system does not appear to be exclusively linked to phylogenetic position, but rather to the physiological adaptation of the species. Histological sections of primary segmental arteries and associated interarterial anastomoses and secondary vessels from the long-finned eel, Anguilla reinhardtii, were examined by light and transmission electron microscopy. Secondary vessels were found to originate from the primary vasculature as depressions through the tunica intima and media, from where they ran perpendicularly to the adventitial layer, before coiling extensively. From here the anastomoses travelled a relatively linear path in the outer margin of the adventitia to re-anastomose with a secondary vessel running in parallel with the primary counterpart. Secondary vessels had a structure quite similar to that of primary vessels; they were lined by endothelial cells on a continuous basement membrane, surrounded by single layer of smooth muscle cells surrounding the vessel. Smooth muscle cells were also found in the vicinity of interarterial anastomoses in the adventitia, but these were more longitudinally orientated. The presence of smooth muscle cells on all aspects of the secondary circulation suggests that this vascular system is regulated in a similar manner as the primary vascular system. Because interarterial anastomoses are structurally integrated with the primary vessel from which they originate, it was anticipated that flow through secondary vessels would to some extent be affected by an increase in primary vascular tone. Immunohistochemical studies showed that primary segmental arteries displayed moderate immunoreactivity to antibodies against 5-hydroxytryptamine and substance P, while interarterial anastomoses and secondary vessels showed dense immunoreactivity. Secondary vessels were followed to the surface of the animal through consecutive sections, where they eventually give rise to capillary beds overlying the scales. Secondary capillary beds were found to supply chloride cells in the skin, suggesting that this vascular system may be involved in cutaneous ionic exchange. Branchial vascular casts from two species of Tetraodontiformes showed that the vessels previously reported as nutrient vessels are with certainty part of the secondary vascular system. In the three-barred porcupine fish, Dicotylichthys punctulatus, interarterial anastomoses originated at high densities from efferent filamental and the efferent branchial arteries, from where they formed progressively larger secondary vessels. Small branches of this vascular system entered the filament body, where it gave rise to numerous side-vessels along the way. Large secondary vessels running in parallel with the efferent branchial arteries were found to constitute an additional arterio-arterial pathway, in that they exited the branchial basket in company with the afferent mandibular artery, the carotid artery and the efferent branchial arteries, from where they gave rise to vascular beds immediately after exit. The secondary vessels in this species were not found to supply the filament musculature; rather this vascular system was supplied by a single vessel derived from the efferent branchial artery, running in parallel with the afferent branchial artery. Secondary vessels were not found on any branchial component in the banded toadfish, Marylina pleurosticta, but in all other aspects the branchial vascular anatomy was similar to that of D. punctulatus. It is proposed that four independent vascular pathways may be present in the teleostean gill. The blood volume and flow rates of the primary (PVS) and secondary vascular system (SVS) were examined in the catadromous euryhaline teleost Lates calcarifer in order to determine whether any of these parameters were subject to change in individuals acclimated to seawater, compared to a group acclimated to freshwater. There was no significant difference in any measured parameter for the two groups. The volumes of the SVS were 0.67 „b 0.13 and 0.76 „b 0.13 mL 100g-1 body mass for FW and SW acclimated animals respectively. This constituted approximately one-third of the total blood volume in both groups. Turnover times for the SVS ranged from 21.0 to 25.2 minutes, demonstrating in accordance with previous publications, that this system is considerably more dynamic than previously assumed.

Vascular casts of three chondrichthian, one dipnoan, one chondrostean and 14 teleostean species were examined by light and scanning electron microscopy in order to give a qualitative and quantitative analysis of interarterial anastomoses (iaas) that indicate the presence (or absence) of a secondary vascular system (SVS). Anastomoses were found to originate from a variety of different primary blood vessels, many of which have not been previously identified as giving rise to secondary vessels. Segmental arteries derived from the dorsal aorta and supplying body musculature were major sites of origin of the SVS, although there was considerable variation in where, in the hierarchy of arterial branching, the anastomoses occurred. The degree of investment in a SVS was species specific, with more active species having a higher degree of secondary vascularisation. This difference was quantified using an absolute count of iaas between Anguilla reinhardtii and Trachinotus baillonii. A range of general features of the SVS is also described. No evidence of iaas was found on the coeliac, mesenteric or renal circulation in any species. Evidence of interarterial anastomoses were lacking in the dipnoan (Sarcopterygii) and chondrichthyan species examined, suggesting that a SVS is restricted to actinopterygian fishes. The presence and distribution of a secondary vascular system does not appear to be exclusively linked to phylogenetic position, but rather to the physiological adaptation of the species. Histological sections of primary segmental arteries and associated interarterial anastomoses and secondary vessels from the long-finned eel, Anguilla reinhardtii, were examined by light and transmission electron microscopy. Secondary vessels were found to originate from the primary vasculature as depressions through the tunica intima and media, from where they ran perpendicularly to the adventitial layer, before coiling extensively. From here the anastomoses travelled a relatively linear path in the outer margin of the adventitia to re-anastomose with a secondary vessel running in parallel with the primary counterpart. Secondary vessels had a structure quite similar to that of primary vessels; they were lined by endothelial cells on a continuous basement membrane, surrounded by single layer of smooth muscle cells surrounding the vessel. Smooth muscle cells were also found in the vicinity of interarterial anastomoses in the adventitia, but these were more longitudinally orientated. The presence of smooth muscle cells on all aspects of the secondary circulation suggests that this vascular system is regulated in a similar manner as the primary vascular system. Because interarterial anastomoses are structurally integrated with the primary vessel from which they originate, it was anticipated that flow through secondary vessels would to some extent be affected by an increase in primary vascular tone. Immunohistochemical studies showed that primary segmental arteries displayed moderate immunoreactivity to antibodies against 5-hydroxytryptamine and substance P, while interarterial anastomoses and secondary vessels showed dense immunoreactivity. Secondary vessels were followed to the surface of the animal through consecutive sections, where they eventually give rise to capillary beds overlying the scales. Secondary capillary beds were found to supply chloride cells in the skin, suggesting that this vascular system may be involved in cutaneous ionic exchange. Branchial vascular casts from two species of Tetraodontiformes showed that the vessels previously reported as nutrient vessels are with certainty part of the secondary vascular system. In the three-barred porcupine fish, Dicotylichthys punctulatus, interarterial anastomoses originated at high densities from efferent filamental and the efferent branchial arteries, from where they formed progressively larger secondary vessels. Small branches of this vascular system entered the filament body, where it gave rise to numerous side-vessels along the way. Large secondary vessels running in parallel with the efferent branchial arteries were found to constitute an additional arterio-arterial pathway, in that they exited the branchial basket in company with the afferent mandibular artery, the carotid artery and the efferent branchial arteries, from where they gave rise to vascular beds immediately after exit. The secondary vessels in this species were not found to supply the filament musculature; rather this vascular system was supplied by a single vessel derived from the efferent branchial artery, running in parallel with the afferent branchial artery. Secondary vessels were not found on any branchial component in the banded toadfish, Marylina pleurosticta, but in all other aspects the branchial vascular anatomy was similar to that of D. punctulatus. It is proposed that four independent vascular pathways may be present in the teleostean gill. The blood volume and flow rates of the primary (PVS) and secondary vascular system (SVS) were examined in the catadromous euryhaline teleost Lates calcarifer in order to determine whether any of these parameters were subject to change in individuals acclimated to seawater, compared to a group acclimated to freshwater. There was no significant difference in any measured parameter for the two groups. The volumes of the SVS were 0.67 „b 0.13 and 0.76 „b 0.13 mL 100g-1 body mass for FW and SW acclimated animals respectively. This constituted approximately one-third of the total blood volume in both groups. Turnover times for the SVS ranged from 21.0 to 25.2 minutes, demonstrating in accordance with previous publications, that this system is considerably more dynamic than previously assumed.

[pt] Segundo relatórios da Organização Mundial da Saúde, as doenças cardiovasculares são a principal causa de óbitos em nível mundial. Podem, porém, ser controladas mediante diagnóstico e tratamento adequados. Nesse contexto, as ferramentas tecnológicas de auxílio ao diagnóstico são importantes para redução do número de óbitos causados por este tipo de doenças. Esta dissertação avalia aos métodos de otimização: Differential Evolution, Generalized Pattern Search, Mesh Adaptive Direct Search e Nelder-Mead Algorithm, aplicados na busca dos parâmetros que modelam a estrutura dos vasos sanguíneos a fim de melhorar os resultados e tempo de processamento da segmentação da árvore vascular em imagens médicas, conforme proposto em (Oliveira, 2013). Neste trabalho, são apresentados conceitos anatômicos e as características das imagens usadas neste estudo. São ainda descritos os métodos de otimização avaliados e a metodologia da segmentação da árvore vascular cujos parâmetros se deseja otimizar. Com essa base, se formula a metodologia de avaliação destes métodos através de uma análise quantitativa, que é produto da formulação de um teste de hipóteses da diferença entre a avaliação média em combinação pareada. Este teste avalia o desempenho dos métodos de otimização quando são aplicados em amostras aleatoriamente escolhidas em cada um dos exames de tomografia computadorizada que pertencem ao banco de dados composto por imagens: sintéticas, coronárias, hepáticas e de fibras nervosas do sistema olfativo. Conforme aos resultados do teste de hipótese, o método de otimização com o melhor desempenho, em acurácia e custo computacional, é escolhido e as conclusões deste trabalho são elaboradas assim como também as propostas de trabalhos futuros nesta mesma linha de pesquisa. / [en] According to the World Health Organization reports, cardiovascular diseases are the worldwide leading cause of death. However, they can be controlled using proper diagnosis tools and treatments. In this way, the diagnosis assisted technological tools are important to reduce the number of deaths caused by this type of diseases. This dissertation assess optimization methods (Differential Evolution, Generalized Pattern Search, Mesh Adaptive Direct Search, Nelder-Mead Algortihm) applied in the search of parameters that model blood vessels structures in order to improve the results and processing time of a vascular tree segmentation method in medical images, as proposed in (Oliveira, 2013). In this work, anatomical concepts and the characteristics of the images used in this study are presented. The optimization methods assessed and the methodology for the segmentation of the vascular tree, whose parameters are to be optimized, are described. Based on that, the procedure to assess different optimization methods is formulated through a quantitative analysis using a hypothesis test formulation of difference between paired means. This test evaluates the performance of the optimization methods using randomly chosen samples in a computerized tomography exams database composed by synthetic, coronary, hepatic and nervous fiber of the olfactory system images. According to the hypothesis test results, the optimization method with the best performance, both in accuracy and computational cost, is selected and the conclusions of this work are elaborated as well as the proposals for further research.

Submitted by Marco Antonio Oliveira da Silva (maosilva@ipen.br) on 2016-12-22T11:54:03Z No. of bitstreams: 0 / Made available in DSpace on 2016-12-22T11:54:03Z (GMT). No. of bitstreams: 0 / A endostatina (ES) é uma proteína inibidora da angiogênese, com ação específica sobre células endoteliais em proliferação, utilizada para tratamento de tumores sólidos. No entanto, o elevado efeito antitumoral da ES observado em animais não é reproduzido em humanos. Com o intuito de potencializar a eficácia terapêutica da ES, produzimos duas proteínas híbridas com dois domínios funcionais. O primeiro domínio é a ES, que apresenta especificidade por células endoteliais ativadas, dirigindo estas proteínas de fusão às células endoteliais em proliferação, promovendo sua internalização e seu efeito inibitório. Como segundo domínio funcional utilizamos os domínios BH3 próapoptóticos de duas proteínas BH3-only com o objetivo de promover a liberação de citocromo C e desencadear o processo de apoptose, aumentando a ação antiangiogênica da ES. Neste trabalho, foram desenhadas duas proteínas de fusão que contêm o domínio BH3 das potentes proteínas pró apoptóticas PUMA e BIM (ES-PUMA e ES-BIM), que deveriam apresentar efeito antiangiogênico potencializado em relação à ES selvagem. A inserção dos fragmentos de DNA codificantes para os domínios BH3 de PUMA e BIM no vetor contendo o gene da ES (pET-ES) foram realizadas por mutagênese sítiodirigida. Estas proteínas de fusão recombinantes foram expressas como corpos de inclusão em E.coli, renaturadas utilizando processo que utiliza alta pressão e purificadas em resina de afinidade por heparina. O tratamento de células endoteliais com as proteínas ES-PUMA e ES-BIM não levou à queda de viabilidade em ensaio de MTS ou de apoptose avaliado por citometria de fluxo, em comparação com os resultados obtidos pelo tratamento com ES. / Dissertação (Mestrado em Tecnologia Nuclear) / IPEN/D / Instituto de Pesquisas Energéticas e Nucleares - IPEN-CNEN/SP

Trinta dias após a indução do diabetes, em aortas com endotélio (E+) de ratas diabéticas (DB), a Rmáx ao cloreto de potássio (KCl) foi reduzida e a resposta à noradrenalina (NA) foi semelhante as controles (CT). A retirada do endotélio (E-) potencializou a resposta ao KCl e NA, porém essa potencialização foi de menor magnitude em DB. A Rmáx à NA em aortas E+: não foi alterada após o seqüestro de ânion superóxido ou inibição da síntese de óxido nítrico (NO) em DB, nas CT esta resposta foi reduzida e aumentada, respectivamente, e foi reduzida apenas nas DB após o bloqueio dos receptores para endotelina (ET). A mobilização de cálcio (Ca2+) em resposta à NA foi reduzida em aorta E+ e E- de DB. Em aortas de ratas diabéticas, as alterações no aparato contrátil, como por exemplo. A redução da mobilização de Ca2+, podem ser responsáveis pela redução da resposta contrátil e a redução da modulação do NO sobre a resposta à NA ou o aumento da liberação de ET pelo endotélio podem ser os responsáveis pela manutenção da resposta à NA. / Thirty days after induction of diabetes, in aortas with endothelium (E+) of diabetic female rats (DB), the maximum response (Rmax) to potassium chloride (KCl) was reduced and the Rmax to noradrenaline (NE) was similar to controls (CT). The endothelium (E-) removal increased the response to KCl and NE, but this increase was of a lower magnitude in DB than in CT. In aortas E+, the Rmax to NA: was not altered by superoxide anion scavenger or nitric oxide (NO) synthesis inhibition in DB, whereas in CT this response is reduced or increased, respectively, and was reduced only in DB after endothelin (ET) receptor blockade. The mobilization of the calcium (Ca2+) in response to NE was reduced in aortas E+ and E- of the DB. In aortas of DB, the alterations in the contractile apparatus, for example, reduction of the Ca2+ mobilization may be responsible for the reduction of the vasoconstriction. The reduction of NO modulation upon the response to NE or the increase of the ET release can be the responsible for the maintenance of the contractile response to NE.

Doenças vasculares são um problema mundial, que representa 28% das mortes no mundo e 66% do total de doenças que acometem os brasileiros. Dessa forma, há um grande interesse em pesquisar formas de prevenção e tratamento dessas doenças. Algumas medidas são relevantes no auxílio de diagnóstico, tal como: tamanho médio dos ramos, diâmetro médio das seções transversais dos vasos e padrões de divisão de ramos. Calcular essas medidas de forma manual é uma tarefa demorada e trabalhosa. Assim, esta Tese tem como objetivo, propor um método computacional de rastreamento e extração de atributos em redes vasculares a partir de imagens 3D de angiografia por ressonância magnética e por tomografia computadorizada. Trata-se de uma abordagem de rastreamento e identificação de bifurcações que difere das técnicas anteriores, utilizando a Transformada de Hough para identificar o diâmetro do vaso em cortes transversais num dado ponto ao longo de um vaso sanguíneo. Mais detalhadamente, essa abordagem utiliza um campo vetorial advindo do cálculo de uma matriz formada por derivadas parciais de segunda ordem, obtida da intensidade luminosa da imagem, para identificar a direção de um ramo de vaso. Além disso, durante o processo de rastreamento de um ramo de vaso, são calculados vários descritores de forma com o objetivo de classificar regiões como pertencentes a uma bifurcação ou não. Em adição a estes descritores, desenvolvemos uma nova medida chamada de variância do raio que permite distinguir, bifurcações, não-bifurcações e segmentos de vaso com stents (aparelho metálico usado para aumentar o diâmetro dos vasos). Para a classificação de bifurcações, criamos a medida de bifurcação, que trata-se de uma combinação linear de todos os descritores de forma apresentados neste trabalho. Testes foram realizados para atestar a eficácia da abordagem proposta, utilizando tanto imagens sintéticas quantoimagens reais. Os resultados mostraram que o método é capaz de rastrear 91% de uma rede vascular sintética variando o ponto de inicialização e 76% variando o nível de ruído. Também foi observado por meio de testes que o método proposto consegue rastrear vasos e identificar bifurcações em imagens reais sem avaliação numérica. Essa abordagem permite a extração da relação hierárquica entre os ramos em uma rede vascular e a extração do padrão de divisão dos vasos, o que contribui sobremaneira para o estudo do comportamento do fenômeno da angiogênese e no auxílio no diagnóstico de anomalias vasculares. / Vascular diseases are a main health problem, representing 28% of deaths worldwide and 66% of all diseases affecting the Brazilian population. Thus, it is important that researches in prevention and treatment of this type of disease increase. Moreover, there are several demands, such as computational tools capable of analyzing and extracting attributes from non-invasive images. The scope of this work is the analysis and extraction of data from magnetic resonance angiography and computed tomography angiography images by highlighting blood vessels. In this context, this thesis aims the development of a novel computational tracking and feature extraction method for vascular networks from 3D images. Our approach presents the following steps: First, identify the vessel cross-sections along it using the Hough transform. Then, compute a matrix composed of second order partial derivatives of image intensity to identify the direction of the vessel. Perform a feature analysis of the vessel contour to classify the bifurcation point, and finally, identify the direction of the new branch in a bifurcation point. The main contribution of this Thesis is the two new measures developed, called radius ratio and bifurcation measure, the radius ratio is capable to distinguish between a region with bifurcation, stents or without both of them. The bifurcation measure is a linear combination that allows to classify a region as bifurcation or not. Tests were performed in order to verify the proposed approach effectiveness, using both synthetic images and real images. The results showed the method is capable to track 91% of synthetic vascular networks varying the seed point and 76% varying the level of noise. Also, we performed tests in real images and by visual evaluation, we could observed that the proposed method was able to track vessels and identify bifurcations from different parts of the body. This approach allows to calculate, in the future, the density of bifurcations in a vascular network, the distance between them, the stenosis and aneurysms grading and characterize specific vessels. In addition, the vascular networks extraction allows the study of the angiogenesis phenomena and vascular anomalies.

A engenharia de tecido pulpar tem como objetivo substituir a polpa dentária inflamada ou necrosada por um tecido saudável e funcional, capaz de formar nova dentina para reparar a estrutura dentária perdida. Assim, os objetivos deste trabalho foram: avaliar a habilidade de diferenciação de célulastronco de dentes decíduos exfoliados humanos (SHED) em odontoblastos funcionais, demonstrando a formação de tecido mineralizado in vivo; e estudar o efeito de VEGF em SHED com relação à estimulação de vias de sinalização celular (STAT3, AKT e ERK), proliferação, migração, formação de estruturas tubulares e diferenciação em células endoteliais. O início do processo de mineralização de SHED tratadas com dexametasona, ácido ascórbico \'beta\' - glicerofosfato pôde ser detectado por meio da produção da enzima fosfatase alcalina a partir da segunda semana de cultura, mas a expressão de RNAm para DSPP só foi observada após 28 dias de indução. Utilizando-se o modelo de fatias de dentes e matrizes condutivas implantadas no dorso de camundongos imunodeprimidos, demonstrou-se a diferenciação de SHED em células semelhantes a odontoblastos, as quais tiveram imunomarcação positiva com o anticorpo DMP-1. A deposição de dentina, seguindo um ritmo centrípeto de crescimento, numa taxa de 14,1 µm por dia também foi demonstrada por meio da marcação com tetraciclina. O tratamento das SHED com VEGF estimulou a fosforilação de ERK e AKT e a diminuiu a fosforilação de STAT3 em um período de uma hora, provavelmente por meio de sua ligação com os receptores VEGFR-1 e NP-1 presentes nestas células. Além disso, VEGF intensificou a organização das SHED em estruturas tubulares, havendo diferença estatisticamente significativa entre os grupos tratado e não tratado a partir do 5o dia de tratamento. Entretanto, VEGF não estimulou a proliferação nem a migração destas células. Os resultados de RT-PCR mostraram que SHED cultivadas em fatias de dentes e matrizes condutivas expressaram VEGFR-2 já após o primeiro dia de estímulo com VEGF. Ademais, os quatro marcadores de células endoteliais (VEGFR-1, VEGFR-2, CD31 e Caderina-VE) foram observados após 21 dias sob estímulo de VEGF, resultado ainda mais evidente aos 28 dias. In vivo, observou-se que SHED transfectadas com o gene LacZ foram capazes de formar estruturas semelhantes a vasos sangüíneos quando implantadas em camundongos, mas a presença de sangue no seu interior não pôde ser observada após 21 dias de implante. Portanto, SHED podem ser estimuladas a se diferenciar em odontoblastos funcionais, capazes de produzir estrutura mineralizada semelhante à dentina. Ademais, VEGF interfere nas vias de sinalização STAT3, ERK e AKT e estimula a formação de estruturas tubulares e a diferenciação de SHED em células endoteliais, mas não a proliferação e migração de SHED. Acreditamos que tecnologia igual ou semelhante à empregada neste estudo poderá eventualmente fornecer ferramentas clínicas para tratamentos endodônticos que visem à regeneração de um tecido pulpar completo e formação de tecido dentinário num futuro não muito distante. / Dental pulp tissue engineering aims to replace the inflamed or necrotic pulp by a healthy and functionally competent tissue able to form new dentin in order to repair lost structure. The purposes of this work were: to evaluate the differentiation ability of stem cells from human exfoliated deciduous teeth (SHED) into functional odontoblasts, showing the formation of mineralized tissue in vivo; and to study the effect of VEGF on SHED with regards to the stimulation of cell signaling pathways (STAT3, AKT and ERK), the proliferation, migration, capillary sprouting, and the differentiation into endothelial cells. The beginning of the mineralization process of SHED treated with dexamethasone, ascorbic acid and beta-glycerophosphate could be detected through the production of alkaline phosphatase after the second week of culture, but the expression of DSPP mRNA was only observed after 28 days of induction. Using the tooth slice and scaffold model implanted in the dorsum of immunocompromised mice, the differentiation of SHED into odontoblast-like cells, which were immunostained with DMP-1 antibody, was demonstrated. Dentin deposition following a centripetal rhythm, in a rate of 14.1 µm per day, was also shown through the tetracycline labeling. VEGF treatment of SHED stimulated the ERK and AKT phosphorilation, and decreased the phosphorilation of STAT3 over 1 hour period, presumably due to its binding to VEGFR-1 and NP-1 receptors in these cells. In addition, VEGF enhanced SHED organization into tubular structures, with statistically significant difference between the treated group and the non-treated one after the 5th day of treatment. However, VEGF did not stimulate proliferation and migration of these cells. RT-PCR results demonstrated that SHED seeded in the tooth slices and scaffolds expressed VEGFR-2 after the first day of VEGF stimulation. Moreover, the four endothelial cell markers (VEGFR-1, VEGFR-2, CD31 and VE-Cadherin) were observed after 21 days of VEGF stimulation, and this result was even clearer after 28 days. In vivo, SHED transduced with LacZ gene were able to give rise to blood vessel-like structures when implanted in immunocompromised mice, but the presence of blood flow was not observed after 21 days of implantation. Therefore, SHED can be stimulated to differentiate into functional odontoblasts, which in turn are able to produce mineralized structure resembling dentin. Furthermore, VEGF interferes with the STAT3, ERK and AKT signaling pathways, and stimulates the formation of tubular structures and the differentiation of SHED into endothelial cells, but does not stimulate SHEDs proliferation and migration. We believe that the same technology employed in this study or a similar one can eventually provide clinical tools for the endodontic treatments aiming at regenerating a complete pulp tissue and forming a dentin tissue in a near future.

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No. of bitstreams: 1 marjoremarcedacostadurao.pdf: 2738553 bytes, checksum: e0f4e4d1fa78e87f2dadbfd8acf3d3c5 (MD5) Previous issue date: 2017-07-18 / Pesquisa delineada no Método Misto operacionalizada em cinco etapas: ambiência, dois estudos secionais e duas abordagens nas representações sociais (estrutural e processual). Objetivou: 1) Avaliar o processo de punção de vasos e trauma vascular periférico na coleta de amostra de sangue na perspectiva dos usuários de uma Unidade de Atenção Primária à Saúde; 2) Compreender as representações sociais elaboradas por usuários a respeito do processo de punção de vasos para coleta de amostra de sangue nas abordagens estrutural e processual e 3) Produzir tecnologia assistencial levedura a respeito da punção de vasos para coleta de amostras de sangue destinadas à realização de exames laboratoriais, com base em evidências locais de boas práticas de punção, e aplicáveis às Unidades de Atenção Primária à Saúde. Foi cenário da investigação uma Unidade de Atenção Primária à Saúde de um município de Minas Gerais. Participaram 417 usuários do SUS, sendo 50 na ambiência; 204 na etapa seccional um e nas abordagens das Teorias das Representações Sociais (59 na Processual e todos na Estrutural) e 163 na etapa seccional dois. Os referenciais teórico-filosóficos foram: Teoria das Representações Sociais, de Leininger e Processo de punção de vasos periféricos na perspectiva de uma atividade comunicacional de conteúdo reificado. Realizaram-se entrevistas individuais, contendo questões abertas, semiabertas e fechadas a partir de instrumento previamente validado, com a técnica de evocações livres a partir de termo indutor. Utilizou-se aplicativo Open Data Kit (ODK) e perguntas guiadas por questões norteadoras. Variáveis quantitativas foram consolidadas em Programa SPSS24 e tratadas por estatística descritiva, conteúdos discursivos analisados no Programa NVivo11 Pro®. e evocações tratadas no Programa EVOC 2000. Foram atendidos requisitos éticos e legais de pesquisa envolvendo seres humanos. Peculiaridades do processo de coleta de amostra de sangue: ser rápido, rotineiro, realizado de forma mecanizada pelos profissionais técnicos de enfermagem e/ou acadêmicos de enfermagem. No cenário, ele ocorreu com as características de: relação profissional/usuário pouco acolhedora, centralizada no procedimento e na tecnicidade do mesmo, impessoal e sem a identificação das necessidades dos usuários. A incidência de trauma vascular periférico no dia da punção foi de 39,3% e de 28,2% dentro das 48h após a realização do procedimento. Duas categorias expressam a origem da construção coletiva dos usuários não puncionados sobre o processo de punção de vasos periféricos: 1) práxis reificadas de punção de veias em si e em outros e 2) práxis reificada de punção e a relação profissional-usuário. O procedimento de punção integra o senso comum dos participantes e é advindo de experiências prévias durante o contato de sua realização com familiares, (des)conhecidos ou vivências próprias. Identificaram-se na abordagem estrutural realizada com usuários não puncionados caráter negativo e ausência de um objeto como componente do núcleo central, retratando uma representação em construção, mesmo diante de procedimento rotineiro para os participantes. Desenvolveram-se Protocolos Assistenciais de orientações destinados aos usuários, profissionais e para avaliação do local de punção. A forma como ocorre o processo de punção de vasos para fins de coleta de sangue requer uma releitura para que se obtenha um cuidado individualizado e capaz de atender às singularidades de enfrentamento do processo de punção de vasos periféricos para fins de coleta de amostra de sangue. O papel do enfermeiro pode impactar a fidedignidade dos resultados laboratoriais e facilitar a atuação da equipe de enfermagem como terapeuta do processo, garantindo o empoderamento do usuário do SUS e a qualidade do cuidado que lhe é prestado no processo de coleta de amostra de sangue para fins laboratoriais. / Research outlined in the Mixed Method Operationalized in five phases: ambience, two sectional studies and two approaches in social representations (structural and procedural). Aimed: 1) To Evaluate the procedure of puncture of vessels and peripheral vascular trauma in the collection of blood samples from the perspective of the users of a Primary Health Care Unit; 2) To understand the social representations elaborated by users regarding the puncture process of blood sampling vessels in the structural and procedural approaches and 3) To produce light-hard assistive technology regarding the puncture of vessels for blood samples collection to perform laboratory tests, based on local evidence of good practices in the puncture process, and applicable to the Units of Primary Health Care. A Primary Health Care Unit of a municipality of Minas Gerais was a research scenario. 417 SUS users participated, 50 in the ambience; 204 in sectional step one and in the approaches of Theories of Social Representations (59 in Processual and all in Structural) and 163 in sectional step two. The theoretical-philosophical references were: Social Representations, Leininger Theories and Peripheral vessel puncture process from the perspective of a communicational activity of reified content. Individual interviews were conducted, containing open, semi-closed and closed questions from a previously validated instrument, with the technique of free evocations from the inducer term. We used the Open Data Kit (ODK) application and questions guided by guiding questions. Quantitative variables were consolidated in Program SPSS24 and treated by descriptive statistics, discursive contents analyzed in the NVivo11 Pro®. Program and evocations treated in the EVOC 2000 Program. Ethical and legal research requirements involving human beings. The specificities of the blood sample collection process were being fast, routine, performed in a mechanized way by nursing technicians professionals and / or nursing students. In the scenario, it occurred with the characteristics of: unfriendly professional / user relationship, centralized in the procedure and in the technicality of the same, impersonal and without the identification of the users' needs. The incidence of peripheral vascular trauma on the day of puncture was 39.3% and 28.2% within 48 hours after the procedure. Two categories express the origin of the collective construction of unpunctured users on the peripheral vessel puncture process: 1) Reified praxis of venipuncture in itself and the others and 2) Reified puncture praxis and the professional-user relationship. The puncture procedure integrates the common sense of the participants and is derived from previous experiences during the contact of their accomplishment with relatives, (un) Known or own experiences. It was identified in the structural approach performed with unpunctured users negative character and absence of an object as a component of the central nucleus, depicting a representation under construction, even before a routine procedure for the participants. Assistance protocols were developed for guidelines for users, professionals and for the evaluation of the puncture site. The way the puncture process of blood vessels takes place requires a re-reading so that individualized care can be obtained and able to meet the singularities of coping with the puncture process of peripheral vessels for the purpose of collecting blood samples. The role of the nurse can impact the reliability of the laboratory results and facilitate the nursing team's performance as a therapist of the process, guaranteeing the empowerment of the SUS user and the quality of the care provided to him in the blood sample collection process for laboratory tests.

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No. of bitstreams: 1 michelenakaharamelo.pdf: 6340172 bytes, checksum: 390b3ea0b5a42d8c811bbac88f7cc6f5 (MD5) Previous issue date: 2017-08-21 / Introdução: O uso dos vasos sanguíneos, apesar de ser reconhecido como útil e benéfico à prática clínica, pode causar iatrogenias, efeitos adversos e comprometer a segurança, o tratamento e o conforto dos usuários. As manifestações de alteração no sítio de inserção dos cateteres intravasculares periféricos (CIVPs) ou nas áreas adjacentes são conhecidas como trauma vascular. Objetivo: produzir tecnologia assistencial leve-dura na modalidade de bundle para subsidiar implementação de intervenções de enfermagem sobre o processo de punção de vasos periféricos e a prevenção de trauma vascular numa Unidade de Pronto Atendimento (UPA) a partir de um diagnóstico situacional, com vistas à estruturação do cuidado de enfermagem baseado em evidências. Métodos e técnicas: Foram referenciais: 1) Políticas Públicas em Urgência/Emergência; 2) Processo de punção de vasos periféricos numa UPA; 3) Teoria das Representações Sociais (Abric) e 4) Teoria de Neuman. Pesquisa de método misto (qualiquanti), operacionalizada em procedimentos sequenciais e simultâneos utilizando intervenções complexas, segundo Medical Research Council (MRC). Cenário: UPA de porte III em Minas Gerais. Foram etapas metodológicas: desenvolvimento (revisão integrativa, ambiência, diário de campo, abordagem estrutural das representações sociais); confiabilidade e pilotagem (coorte e diário de campo); avaliação (construção do bundle, processo educativo e diário de campo) e implementação (educação permanente com segmento e monitoramento e diário de campo). Participaram 56 membros da equipe de enfermagem e 228 usuários do Sistema Único de Saúde (SUS) que tiveram suas veias puncionadas avaliadas numa coorte (252). Dados coletados por triangulação de técnicas e métodos (semiológicos, fotográficos, entrevistas, técnicas projetivas, observação participativa e consulta a acervo eletrônico) com apoio do aplicativo Open Data Kit (ODK). Foram utilizados programas EVOC 2000® e SPSS24®, sendo atendidos requisitos éticos/legais de pesquisa envolvendo seres humanos. Resultados: usuários se apresentaram como homens (58,3%), de baixa escolaridade (56,8%), idosos (55,6%) e alocados no setor amarelo (67,1%); incidência de trauma vascular periférico de 62,3% (predominando endurado, edema e dor) documentados por critérios imagéticos. Houve correlação (p-valor= 0,001-0,040) entre sexo, faixa etária, hipertensão, sangue visível e autoavaliação de ter veia difícil de puncionar com a variável de desfecho (trauma vascular). Produziuse tecnologia assistencial leve-dura na modalidade de bundle, alicerçado em diagnóstico situacional e evidências científicas. Foram utilizadas ferramentas tecnológicas/comunicacionais e didático-pedagógicas (vídeo e protocolo) para subsidiar abordagens individuais e grupais. A revisão integrativa delineou acervos sobre tipos/causas de trauma vascular e intervenções compatíveis com usuários de uma UPA, explicitando perfis modificáveis/não modificáveis por intervenções de enfermagem. Elementos simbólicos retrataram respostas humanas dos usuários e profissionais que norteiam demandas de cuidados e lacunas na percepção de como os mesmos concebem, enfrentam, valoram, experienciam a temática e lidam com ela. Concepção coletiva dos profissionais alicerçada numa técnica fragmentada desconectada da relação dialógica sensível e singular que deve presidir qualquer ato profissional. Conclusões/Considerações finais: a complexidade da temática e o distanciamento do enfermeiro de seu protagonismo no direcionamento da atuação da equipe de enfermagem e das atividades assistências/gerenciais possibilitaram negar a hipótese de nulidade de H1 e a hipótese de nulidade de H2. Foram contribuições: diagnóstico situacional, criação do bundle e abordagem educativa. Recomenda-se: investimento institucional em educação permanente da equipe de enfermagem e de materiais apropriados, criação de um protocolo institucional de notificação de eventos adversos e realização de uma coorte após a implementação do bundle e a replicação da investigação em outras UPAs. / Introduction: The use of blood vessels, despite being recognized as useful and beneficial to clinical practice, can cause iatrogenic, adverse effects and compromise the safety, treatment and comfort of the users. The manifestations of change in the insertion site of the peripheral intravascular catheters or adjacent areas are known as vascular trauma. Objectives: to produce light-hard assistance technology in the bundle modality to support the implementation of nursing interventions on the peripheral vessel puncture process and prevent the vascular trauma in an Emergency Department (ED) based on a situational diagnosis, with a view to the structuring of evidence-based nursing care. Methods and techniques: Reference was made to: 1) Public Policies in Emergency; 2) Procedure of puncture of peripheral vessels in a UPA; 3) Theory of Social Representations (Abric) and 4) Theory of Neuman. Mixed-method research, operationalized in sequential and simultaneous procedures using complex interventions, according to the Medical Research Council (MRC). Scenario: UPA of size III in Minas Gerais. There were methodological steps: Development (integrative review, ambience, field diary, structural approach to social representations); Reliability and Piloting (cohort and field diary); Evaluation (bundle construction, educational process and field diary) and Implementation (permanent education with segment and monitoring and field diary). Participants were 56 members of the nursing team and 228 users of the Sistema ùnico de Saúde (SUS) who had their punctured veins evaluated in a cohort (252). Data collected by triangulation of techniques and methods (semiological, photographic, interviews, projective techniques, participatory observation and electronic collection) with the support of the Open Data Kit (ODK), EVOC 2000® and SPSS24® programs were used, and ethical/legal research requirements involving human beings were met. Results: users presented as: men (58.3%), with low schooling (56.8%), elderly (55.6%) and allocated in the yellow sector (67.1%); incidence of peripheral vascular trauma of 62.3% (predominantly enduring, edema and pain) documented by imaging criteria. There was a correlation (p-value = 0.001-0.040) between sex, age, hypertension, visible blood and self-evaluation of having a difficult-to-puncture vein with outcome variable (vascular trauma). Light-hard care technology was developed in bundle modality, based on situational diagnosis and scientific evidence. Technological / communicational and didactic-pedagogical tools (video and protocol) were used to subsidize individual and group approaches. The integrative review delineated collections on types/causes of vascular trauma and interventions compatible with ED users, explaining modifiable/non-modifiable profiles by nursing interventions. Symbolic elements portrayed human responses from users and practitioners who drive care demands and gaps in the perception of how they conceive, confront, value, experience, and deal with the issue. Collective conception of professionals based on a fragmented technique disconnected from the sensitive and singular dialogical relationship that should preside any professional act. Conclusions / final considerations: the complexity of this issue and the distance of the nurses from their role in directing the nursing staff and assisting/management activities made it possible to deny the hypothesis of nullity of H1 and the hypothesis of nullity of H2. Contributions were: situational diagnosis, bundle creation and educational approach. It is recommended: institutional investment in continuing education of the nursing team and appropriate materials, creation of an institutional protocol for reporting adverse events and completion of a cohort after the implementation of the bundle and replication of this research in other EDs.

The present study was undertaken to evaluate the rat as a model for middle ear re­search. The rat was chosen primarily because the gross structure of its middle ear shows several similarities to that of man. It was considered of great importance to make a thorough structural study of the rat middle ear and to compare the results with those reported for the human middle ear. The thesis therefore includes indepen­dent studies on various aspects of rat middle ear structure and function as well as a review of the literature. The most pertinent findings in the experimental part of this study were the following. The rat Eustachian tube consists of a nasopharyngeal, and a cartilaginous and bony portion. The orifice of the nasopharyngeal portion is composed of two soft tissue lips, which appear to be opened mainly by the action of the salpingopharyngeal mus­cle, but also by the levator and tensor veli palatini muscles. The cartilaginous por­tion appears to be opened solely by the tensor veli palatini muscle. The tensor tympani muscle seems to have no effect on the tube. A ciliated and secretory epithelium lines the inferomedial walls of the tube throughout its length. In the tympanic cavity these thelial cell types extend as two tracts - one anterior and the other inferoposterior to  the promontory - which communicate with the epitympanic/attic compartments. The remaining parts of the tube and the tympanic cavity are covered by a squamous/cuboidal, non-ciliated epithelium. The subepithelial loose connective tissue contains vessels, nerves, and connective tissue cells, among these mast cells. The mast cells are confined to areas covered by the ciliated epithelium, and in the floor of the bulla, in the pars flaccida, and along the manubrial vessels. Glands are restricted to the Eustachian tube. In the clearance/transport of serum-like material, from the epitympanum towards the tube, hydrostatic forces appear to be important. The tympanic membrane is vascularized from meatal and tympanal vessels. Meatal ves­sels branch in the pars flaccida and along the handle of the malleus, where they are localized directly beneath the outer, keratinizing, stratified, squamous epithelium. Furthermore, meatal vessels form a vascular network at the junction between the fi­brocartilaginous annulus and the tympanic sulcus. Tympanal vessels send branches to the periphery of the pars tensa, where they run immediately beneath the tympanal, simple, squamous epithelium. In the major portion of the pars tensa, no blood vessels were found. The rat stapedial artery is a thin-walled vessel with a wide lumen. Without branch­ing, it runs through the tympanic cavity to the extratympanal regions it supplies. In contrast to the corresponding artery in man, the rat stapedial artery persists throughout life. The artery does not seem to be affected by the fluid produced during experimentally induced otitis media with effusion. The middle ear structure in the rat and in man show both similarities and differ­ences. If the differences are kept in mind and considered, it would seem that the rat is indeed a suitable model for experimental middle ear research. / digitalisering@umu