Introdução: A indicação de endarterectomia carotídea (EAC) profilática em conjunto com revascularização miocárdica (CRM) permanece assunto indefinido. A cirurgia conjunta é amplamente difundida, mas seus resultados vêm sendo questionados. Método: Coorte retrospectiva de 691 pacientes submetidos à CRM, escolhidos aleatoriamente. Avaliação realizada para dados gerais, presença de lesão carotídea, ateromatose aórtica, desfechos neurológicos e óbito. Resultados: Dentre 691 pacientes submetidos à CRM, 16 pacientes apresentaram acidentes vasculares cerebrais (AVC). Dentre esses, 11 pacientes (68.75%) apresentaram AVC localizados em áreas não compatíveis com as lesões carotídeas, sendo três deles com lesões calcificadas na aorta ascendente. Os pacientes com estenose carotídea apresentaram taxa similar de eventos neurológicos totais, AVC e óbito, comparados com pacientes sem estenose carotídea. Um subgrupo de 35 pacientes com estenose carotídea foi submetido à cirurgia coronariana com (14 pacientes) ou sem (21 pacientes) cirurgia de carótida, obtendo-se taxa de eventos neurológicos totais, AVC e óbito estatisticamente semelhantes. Os pacientes com calcificações aórticas apresentaram risco maior de eventos neurológicos (14,58% versus 6.55%, p=0.011), AVC (3,12% versus 2,18%, p=0,47) e óbito (8,33% versus 4,37%, p=0.12). Discussão: Os eventos neurológicos após CRM correlacionam-se com ateromatose aórtica. Os AVC freqüentemente não têm relação linear com a estenose carotídea. Estratégias para minimizar embolia da aorta podem diminuir as taxas de intercorrências neurológicas. / Introduction: The management of patients with simultaneous disease of carotid and coronary arteries is controversial. Studies showed that aortic calcifications might play a role in postoperative stroke at coronary artery bypass graft (CABG), carotid lesions may not be as important as previously considered. Method: A retrospective cohort of a randomly selected group (including elective and emergency operations) of 691 patients submitted to CABG was reviewed for general data, neurological complications and mortality. Results: Among 691 CABGs 16 patients had postoperative stroke. Among these, 11 patients (68.75%) had strokes not matching carotid lesions and anatomic presentation, three of those had detectable aortic calcifications. The patients with critical carotid stenosis had similar rates of neurological events, stroke and death as compared to patients without. The patients with aortic calcifications presented a higher risk of neurological events (14.58% versus 6.55%, p=0.011), stroke (3.12% versus 2.18%, p=0.47) and death (8.33% versus 4.37%, p=0.12). Discussion: The postoperative neurological events after CABG can be related to aortic calcifications. The strokes after coronary bypass may occur independently of the carotid lesions. Strategies to prevent aortic emboli may help preventing many post-operative strokes.

A hipertrofia cardíaca é um mecanismo de adaptação do coração ao aumento de demanda. De acordo com o estímulo, fisiológico ou patológico, a hipertrofia apresenta diferentes características morfológicas e moleculares. Compreender os mecanismos comuns e distintos entre os dois tipos de hipertrofia é um passo importante para o desenvolvimento de estratégias de prevenção e tratamento da IC. Dentre os mecanismos distintos cabe ressaltar a participação das espécies reativas do oxigênio (EROs) que parecem estar presentes em altos níveis na hipertrofia cardíaca patológica e em baixos na fisiológica. Além disso, o papel regulatório dos microRNAs (miRs) tem sido demonstrado nas doenças cardiovasculares. No entanto, a influência das EROs no desenvolvimento da hipertrofia e nas adaptações decorrentes a ela ainda não está estabelecido. Assim, nosso objetivo foi avaliar as diferenças morfológicas e moleculares da hipertrofia cardíaca fisiológica, induzida pelo exercício, e da patológica, induzida por bandeamento aórtico (TAC), e sua modulação pela vitamina E. Os modelos de exercício e TAC desenvolveram hipertrofia cardíaca de forma compatível com o estímulo recebido. Essas adaptações ocorreram conjuntamente com alterações na expressão dos miRs-21, -26b, -150, -210 e -499. A vitamina E inibiu o estímulo angiogênicos, no modelo fisiológico, assim como a expressão dos miRs-21, -150 e -210. No entanto, esses efeitos não alteraram o fenótipo final da hipertrofia cardíaca fisiológica. No modelo patológico, por outro lado, a vitamina E reduziu a fibrose e o dano oxidativo, além de alterar a expressão de miRs já descritos no desenvolvimento da hipertrofia cardíaca patológica. Novamente, esse efeito não foi suficiente para reduzir a hipertrofia cardíaca. Em conjunto, os dados desse estudo sugerem que a vitamina E e/ou sua capacidade antioxidante têm a capacidade de influenciar de forma benéfica a hipertrofia patológica; no entanto, seus efeitos podem ser desfavoráveis no estímulo fisiológico. / Cardiac hypertrophy is an adaptive mechanism of the heart to the increased demand. According to the stimulus, physiological or pathological, cardiac hypertrophy present different morphological and molecular features. Understanding both the unique and the shared features in each type of hypertrophy is an important step to the development of novel approaches in the HF management. Among the unique mechanisms, the participation of reactive oxygen species (ROS) seems to be present at high levels in pathological and at low levels in physiological cardiac hypertrophy. Furthermore, the regulatory role of microRNAs (miRs) have been shown in cardiovascular diseases. However, ROS influence in cardiac hypertrophy development and their adaptations were not established yet. Thus, our objective was to evaluate morphological and molecular differences between physiological cardiac hypertrophy (physical exerciceinduced) and pathological cardiac hypertrophy (transverse aortic constrictioninduced), and its modulation by vitamin E. Exercise and TAC models developed cardiac hypertrophy in a manner consistent with the received stimulus. These adaptations occurred along with changes in miR-21, -26b, -150, -210 and -499 expression. Vitamin E inhibited angiogenic adaptations, as well as miR-21, -150 and -210 expression in physiological model. However, these effects did not change the final physiological cardiac hypertrophy phenotype. On the other hand, in the pathological model, vitamin E reduced oxidative damage and fibrosis, and altered the expression of miRs described in pathological cardiac hypertrophy development. Again, this effect was not sufficient to reduce cardiac hypertrophy. In conclusion, vitamin E and/or its antioxidant capacity have the capacity to influence the pathological hypertrophy in a beneficial way, but its effects can be unfavorable in the physiological stimulus.

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Simona Rudišarová Supervisor: PharmDr. Marie Vopršalová, CSc. Title of diploma thesis: The effect of quercetin metabolite on the rat isolated arteries This study is focused on the research of the effects of flavonol quercetin and its selected metabolites (3-(3-hydroxyphenyl)propionic acid and 3-hydroxybenzoic acid) on blood vessels in vitro. A standardized method of an isolated rat aorta was used. The aim of this study was to find out how substances act on the isolated aorta and if the effect of the effective metabolite is dependent on the presence of the endothelium in the vessel. After precontracting with noradrenaline, individual concentrations of the test substances were added to the aortic ring bath (final concentrations in the bath ranging from 10-7 to 10-3 M). From the measured values EC50 values (quercetin = 3,63 . 10-5 M, 3-(3-hydroxyphenyl)propionic acid = 3,21 . 10-6 M, 3-hydroxybenzoic acid = 4,38 . 10-5 M) were obtained and the corresponding DRC curves were constructed using S.P.E.L. Advanced Kymograph Software. The experiment was performed once again with quercetin and 3-(3-hydroxyphenyl)propionic acid on the endothelium-denuded vessel. According to the results is...

O expressivo número de brasileiros que necessitam correção anatômica da estenose valvar aórtica acentuada e que não realizam cirurgia de substituição valvar devido ao risco proibitivo justifica a necessidade de investigação, tanto da efetividade no cenário clínico real quanto dos custos impostos ao Sistema Único de Saúde e aos planos de saúde suplementar brasileiros pela incorporação do implante transcateter de valva aórtica, que tem se demonstrado efetivo mas oneroso, internacionalmente. No primeiro artigo da tese, avaliaram-se os desfechos intra-hospitalares, a sobrevida e o reembolso pela internação hospitalar de 41 pacientes com idade média de 78,7 ± 6,3 anos, estenose valvar aórtica acentuada, com recusa cirúrgica e decisão multidisciplinar por tratamento transcateter entre outubro de 2010 e outubro de 2015. Os sujeitos foram seguidos prospectivamente por um período mediano de 15,2 (4,5 – 25,6) meses e a sobrevida estimada em 1 e 2 anos foi de 73,2% e 64,1%, respectivamente. Identificou-se que hipertensão pulmonar e revascularização miocárdica cirúrgica prévia estavam independentemente associadas à menor sobrevida. O valor mediano reembolsado pelos pacientes atendidos pelo Sistema Único de Saúde foi R$ 108.634,34 (101.051,05 – 127.255,27) e R$ 115.126,77 (94.603,21 – 132.603,01) para aqueles internados por planos de saúde suplementar ou particulares, sendo o respectivo valor mediano reembolsado pela prótese valvar de R$ 82.000,00 (82.000,00 – 95.450,00) e 84.050,00 (75.000,00 – 92.400,00) Em um grupo de 585 procedimentos de troca valvar aórtica cirúrgica em indivíduos com idade ≥ 60 anos, realizados entre janeiro de 2010 e dezembro de 2015 na mesma instituição, a mortalidade intra-hospitalar estava associada à idade e foi de 5,9% naqueles com idade entre 60 e 70 anos, 10,8% entre 70 e 80 anos e de 22,2% ≥ 80 anos. O reembolso mediano foi de R$ 14.035,96 (11.956,11 – 16.644,90) para os internados pelo Sistema único de Saúde e R$ 20.273,97 (15.358.03 – 32.815,49) pelos planos de saúde suplementar ou particulares. No segundo artigo da tese, identificou-se que do total de 819 pacientes consecutivamente incluídos no Registro Brasileiro de Implante de Bioprótese Aórtica por Cateter entre janeiro de 2008 e outubro de 2015, 15 (1,8%) sofreram perfuração do ventrículo esquerdo. Os pacientes que apresentaram perfuração eram mais idosos (85,4 ± 6,3 vs. 81,5 ± 7,3 anos, p=0,038), predominantemente mulheres (80,0% vs. 50,5%, p=0,024), apresentavam maior fração de ejeção (67,3 ± 7,8% vs. 58,6 ± 15,0%, p=0,001), menor massa ventricular esquerda (203,9 ± 47,1g vs. 247,6 ± 78,7g, p=0,039) e menor altura do tronco da coronária esquerda (11,2 ± 5,4mm vs. 14,0 ± 3,3mm, p=0,034). Os preditores independentes de perfuração do ventrículo esquerdo foram idade e fração de ejeção. No terceiro artigo, descreve-se um caso de ablação septal para tratamento de miocardiopatia hipertrófica obstrutiva assimétrica para posterior implante transcateter de valva aórtica, sugerindo que esta seja uma estratégia factível quando da concomitância dessas duas condições Em conclusão, os desfechos do tratamento transcateter da estenose valvar aórtica acentuada em pacientes inoperáveis são compatíveis com aqueles do cenário idealizado dos ensaios clínicos randomizados, apesar de estarem associados a custos maiores que os anteriormente estimados por painéis de especialistas. O tratamento cirúrgico, por sua vez, apresentou mortalidade maior que aquela idealizada ou relatada como usual. A hipercinesia do ventrículo esquerdo pode favorecer o trauma determinado pelo guia metálico, posicionado em seu interior para realização do procedimento, estando a fração de ejeção independentemente associada à chance de perfuração. Ainda, a ablação septal por álcool eletiva, anterior ao implante transcateter da valva aórtica, é uma abordagem factível para pacientes com hipertrofia ventricular esquerda assimétrica obstrutiva associada à estenose valvar aórtica. / The expressive number of Brazilians who require an anatomic correction for severe aortic valve stenosis and who do not undergo valvar replacement surgery due to prohibitive risk justifies the need to investigate both the effectiveness in the real clinical scenario and the costs imposed to the Public Health System and the Supplementary Health System for the incorporation of the transcatheter aortic valve implantation, which has been shown to be effective but onerous, internationally. In the first article of the thesis, the in-hospital outcomes, long-term survival and reimbursement for 41 patients, with a mean age of 78.7 ± 6.3 years, sever aortic valve stenosis, with surgical refusal and multidisciplinary decision for transcatheter treatment, between October 2010 and October 2015 are described. Subjects were prospectively followed for a median period of 15.2 (4.5 - 25.6) months and the estimated survival at 1 and 2 years was 73.2% and 64.1%, respectively. It was identified that pulmonary hypertension and previous coronary artery bypass grafting were independently associated with lower survival. Median reimbursed values by the Public Health System was R$ 108,634.34 (101,051.05 - 127,255.27) and by supplementary health plans was R$ 115,126.77 (94,603.21 - 132,603.01). The respective median values reimbursed for the valve prosthesis was R$ 82,000.00 (82,000.00 - 95,450.00) and 84,050.00 (75,000.00 - 92,400.00) In a group of 585 surgical aortic valve replacement procedures in subjects aged ≥ 60 years, performed between January 2010 and December 2015 in the same institution, in-hospital mortality was associated with age and was 5.9% in those with age between 60 and 70 years, 10.8% between 70 and 80 years and 22.2% in ≥ 80 years. The median reimbursement was R$ 14,035.96 (11,956.11 - 16,644.90) for those hospitalized by the Public Health System and R$ 20,273.97 (15,358.03 - 32,815.49) by supplementary or private health plans. In the second article of the thesis, it was identified that of the total of 819 patients consecutively included in the Brazilian Registry of Aortic Bioprosthesis Implantation by Catheter (RIBAC) between January 2008 and October 2015, 15 (1.8%) suffered perforation of the left ventricle. Patients with perforation were older (85.4 ± 6.3 vs. 81.5 ± 7.3 years, p=0.038), predominantly women (80.0% vs. 50.5%, p=0.024), had a higher ejection fraction (67.3 ± 7.8% vs. 58.6 ± 15.0%, p=0.001), lower left ventricular mass (203.9 ± 47.1g vs. 247.6 ± 78, 7g, p=0.039) and shorter distance between the aortic annulus and the left main coronary artery ostium (11.2 ± 5.4mm vs. 14.0 ± 3.3mm, p=0.034). The independent predictors of left ventricular perforation were age and ejection fraction. In the third article, a case of septal ablation was described for the treatment of asymmetric obstructive hypertrophic cardiomyopathy for posterior transcatheter aortic valve implantation, suggesting that this is a feasible strategy when these two conditions are concomitant In conclusion, the outcomes of transcatheter treatment of severe aortic stenosis in inoperable patients are compatible with those in the ideal scenario of randomized clinical trials, although they are associated with higher costs than previously estimated by expert panels. Surgical treatment, on the other hand, presented higher mortality than that idealized or reported as usual. The left ventricle hyperkinesia may favor the trauma determined by the metallic guide, positioned inside it to perform the procedure, the ejection fraction being independently associated with the chance of perforation. Furthermore, elective alcohol septal ablation, prior to transcatheter aortic valve implantation, is a feasible approach for patients with obstructive asymmetric left ventricular hypertrophy associated with aortic valve stenosis.

A hipertrofia cardíaca é uma adaptação do coração frente a estímulos de crescimento, sejam eles patológicos e irreversíveis como a sobrecarga de pressão ou de volume, ou fisiológicos e reversíveis como a gravidez e o exercício físico. A hipertrofia derivada de estímulos patológicos é conhecida como mal adaptativa enquanto que a hipertrofia proveniente de estímulos ditos fisiológicos é conhecida como benéfica ou adaptativa. Embora ambas hipertrofias tenham fatores em comum no que diz respeito ao crescimento do cardiomiócito e adaptações moleculares, elas acabam divergindo para desfechos completamente diferentes. A hipertrofia patológica evolui para um quadro de disfunção cardíaca ao passo que a hipertrofia fisiológica não acarreta nenhum dano funcional ao miocárdio. Essa linha tênue entre um fenótipo e outro envolve mecanismos celulares complexos que ainda precisam ser esclarecidos. Dentro deste cenário, os microRNAs aparecem como reguladores de diversos processos celulares, e têm sido associados ao crescimento miocárdico. Portanto, nosso objetivo foi comparar o padrão de expressão de microRNAs entre os modelos de hipertrofia fisiológica, induzido por natação (SWIM), e o modelo de hipertrofia patológica, induzida por bandeamento aórtico transtorácico (TAC). As análises foram realizadas após 28 dias para o modelo de natação, e 35 dias para o modelo de TAC. A comparação foi realizada através da técnica de microarranjo de microRNAs (Affymetrix). Interessantemente, apenas 20 microRNAs apresentaram níveis de expressão distinta entre os dois modelos de hipertrofia. Destes, 12 microRNAs apresentaram aumento de expressão (miR-193a-3p, miR-299a-5p, miR- 127-5p, miR-214-5p, miR-188-5p, miR-326-3p, miR-6395, miR-547-3p, miR-199a-5p, miR-381-3p, miR-223-3p e miR-199b-5p) e 8 estavam com seus níveis diminuídos (miR11 708-5p, miR-30c-1-3p, miR-22-5p, miR-6921-5p, miR-30a-3p, miR-30e-3p, miR-27a-5p and miR-6975-5p) no grupo TAC em relação ao grupo SWIM. Além disso, apenas 3 microRNAs, miR-21a-5p, miR-206-3p e miR-1983, apresentaram aumento de expressão tanto no grupo TAC quanto no grupo SWIM em comparação aos grupos SHAM e Sedentário, respectivamente. Após isso, foi realizada uma busca por possíveis alvos destes microRNAs na base de dados KEGG Pathway que identificou 4 rotas enriquecidas (665 genes) entre os alvos dos microRNAs reduzidos, e 80 rotas (3394 genes) fortemente associadas aos microRNAs que estavam aumentados no grupo TAC comparado ao SWIM. Conclui-se que existem microRNAs específicos para o desenvolvimento da hipertrofia cardíaca fisiológica, bem como patológica conforme os dados obtidos na análise de microarranjo. Além disso, os possíveis alvos destes microRNAs parecem estar envolvidos em rotas bastante envolvidas no crescimento celular, sobrevivência e adaptação cardíaca. / Cardiac hypertrophy is a heart adaptation in response to growth stimuli whether pathological and irreversible such as pressure overload or physiological and reversible as pregnancy and exercise. Hypertrophy because of pathological stimuli is known as mal adaptive while the one that comes from physiological triggers is known as beneficial or adaptive. Although both have similarities about cardiomyocyte growth and molecular adaptations, they diverge to distinct outcomes. The pathological hypertrophy evolves to a pattern of cardiac dysfunction while the physiological one does not cause any damage to the heart. This tenuous line between those phenotypes involves complex cellular mechanisms that need to be clarified. In this context, microRNAs are considered as regulators of many biological processes, and have been associated to myocardial growth. Therefore, our aim was to compare microRNA expression between physiological (swiminduced) and pathological (TAC-induced) hypertrophy. The analysis was performed after 28 days for SWIM protocol and 35 days for TAC model. The comparison was done using microRNA microarray technology (Affymetrix). Interestingly, only 20 microRNAs were differential expressed between both models. Out of those, 12 were up regulated (miR- 193a-3p, miR-299a-5p, miR-127-5p, miR-214-5p, miR-188-5p, miR-326-3p, miR-6395, miR-547-3p, miR-199a-5p, miR-381-3p, miR-223-3p and miR-199b-5p) while 8 were down regulated in TAC group compared to SWIM group. Besides, only 3 microRNAs, miR-21a-5p, miR-206-3p and miR-1983, were upregulated in TAC and SWIM model compared to SHAM and SED groups. After that, a search at KEGG Pathway database retrieved 4 pathways (665 genes) enriched with targets from microRNAs downregulated and 80 pathways (3394 genes) enriched with targets from up-regulated microRNAs in in 13 TAC group compared to SWIM group. In conclusion, there are microRNAs specific committed to the physiological cardiac hypertrophy development as well to the pathological cardiac growth as observed in our microarray data. Furthermore, the possible targets of those microRNAs could be involved in pathways associated with cellular growth, survival and cardiac adaptation.

Définie comme étant le rétrécissement de la valve, la sténose aortique (SA) est la 3ème pathologie cardiovasculaire dans les pays industrialisés. Touchant essentiellement les personnes âgées de plus de 65 ans, cette pathologie représente un véritable problème de santé publique compte tenu du vieillissement de la population. Considérée initialement comme issue d’un processus passif de dégénérescence, il est désormais établi que la sténose aortique est une pathologie dite « atherosclerosis-like » caractérisée par les processus d’inflammation, de fibrose, de néo-angiogenèse et de calcification. Certaines protéines de la voie de coagulation tel que le facteur tissulaire (FT) sont connues pour avoir un rôle pro-fibrotique et participent activement au développement des lésions athéroscléreuses. Leurs rôles dans la SA semblent donc probables et restent à être identifiés.Composante cellulaire majeure de la valve aortique, les VICs présentent 5 sous-populations distinctes : les cellules progénitrices embryonnaires (EPCs), les cellules progénitrices (pVICs), quiescentes (qVICs), activées (aVICs) et ostéoblastiques (obVICs). Au cours de la valvulogenèse, les EPCs permettent la cellularisation de la valve en se différenciant en qVIC. Celles-ci maintiennent l’homéostasie valvulaire et, en cas de lésion, s’activent (aVICs) pour réparer efficacement le tissu valvulaire. L’inflammation valvulaire et l’activation des VICs initient la sécrétion de protéines pro-calcifiantes induisant la différenciation des aVICs en obVICs. Enfin, les pVICs, naturellement présentes au sein de la valve (appelées résidantes) ou issues de la circulation sanguine (appelées hématopoïétiques), semblent favoriser le renouvellement cellulaire et peuvent être impliquées dans les processus angiogénique et ostéoblastique.Bien que décrites, la validation de la culture primaire des VICs par le suivi de ces sous-populations n’avait pas été réalisé et à constituer notre premier objectif. Nous avons ensuite étudié l’implication des voies de signalisation du FT dans le développement de la SA.Dans le cadre du suivi longitudinal des VICs depuis les valves aortiques humaines contrôles et pathologiques jusqu’à la culture in vitro réalisée sur plastique et sur collagène, nous avons tout d’abord montré que les différentes sous-populations étaient présentes au sein de ces valves avec des localisations et des proportions différentes selon l’état physiopathologique du tissu. Après digestion enzymatique de la valve, elles sont toutes retrouvées mais lors de la mise en culture, les pVICs hématopoïétiques ont disparu, quel que soit le support. Nous avons ainsi validé le modèle de culture primaire des VICs tout en mettant en lumière ses limites : absence des pVICs hématopoïétique, activation et différenciation ostéoblastique spontanée des VICs au cours de la culture.Dans le cadre de l’’étude de l’implication du FT dans le développement de la SA, nous avons montré sa colocalisation avec la thrombine et les calcifications de valves pathologiques. A partir de la culture primaire de VICs issues de valves humaines contrôles et pathologiques, nous avons montré que l’expression et l’activité du FT étaient constitutivement plus importantes pour les VICs pathologiques et que son expression pouvait être induite par l’IL1β. De plus, l’activation du FT, en présence de son ligand le facteur VII, induit, directement et via le récepteur PAR2, différentes voies de signalisation impliquées dans la prolifération cellulaire et les processus de fibrose et de calcification. Cette étude suggère ainsi que le FT produit par les VICs est un médiateur clef dans le développement de la sténose aortique. / Defined as the narrowing of the aortic valve, aortic stenosis (AS) is the third cardiovascular pathology in industrialized countries. Affecting mainly people aged over 65 years, AS represents a major public health problem because of the aging of the population. After initially been considered as a passive degenerative process, it is now established that AS is an "atherosclerosis-like " disease characterized by the processes of inflammation, fibrosis, neo-angiogenesis and calcification. Some proteins of the coagulation pathway such as tissue factor (TF) are known to have a pro-fibrotic role and actively participate in the development of atherosclerotic lesions. Their implication in AS seems, therefore, probable and remain to be identified.Prevalent cellular component of the aortic valve, VICs have five distinct subpopulations: embryonic progenitor cells (EPCs), progenitor cells (pVICs) quiescent (qVICs), activated (aVICs) and osteoblastic (obVICs). During the valvulogenesis, EPCs allow the cellularization of the valve, differentiating into qVICs. These cells maintain the valvular homeostasis and, in case of damage, are activated (aVICs) to effectively repair the valve tissue. The valvular inflammation and VICs activation initiate the secretion of pro-calcifying proteins inducing the differentiation of aVICs into obVICs. Finally, pVICs, naturally present within the valve (called resident) or from the blood circulation (called hematopoietic), seem to promote cell renewal and may be involved in the angiogenic and osteoblastic processes.Although described, these subpopulations have never been studied longitudinally, in respect to their behavior in vitro. Our first objective was to perform this investigation. Our second objective was to study the potential role of TF pathway in the deleterious mechanisms of AS.As part of the longitudinal follow-up of VICs from control and pathological human aortic valves to the in vitro culture performed on plastic and collagen, we first showed that different subpopulations were present in these valves with different locations and proportions according to the pathophysiological state of the tissue. After enzymatic digestion, all subpopulations are found but, in culture, hematopoietic pVICs disappeared, whichever the support. Thus, we validated the primary culture model of VICs while highlighting its limitations: lack of hematopoietic pVICs, spontaneous osteoblastic differentiation and activation of VICs in culture.As part of the study the involvement of FT in the AS development, we showed its colocalization with thrombin and calcifications of pathological valves. We showed that the expression and activity of TF were constitutively more important in VICs from fibrocalcified valves than control ones and that IL-1β for pathological VICs and that its expression could be induced by IL1 beta. In addition, TF activation in the by its ligand FVII, induced, directly and via the PAR-2 receptor, different signaling pathways involved in cell proliferation and the processes of fibrosis and calcification. Thus, our findings suggest that the FT expressed by VICs mediates fibrocalcific processes of aortic stenosis.

Introdução: A indicação de endarterectomia carotídea (EAC) profilática em conjunto com revascularização miocárdica (CRM) permanece assunto indefinido. A cirurgia conjunta é amplamente difundida, mas seus resultados vêm sendo questionados. Método: Coorte retrospectiva de 691 pacientes submetidos à CRM, escolhidos aleatoriamente. Avaliação realizada para dados gerais, presença de lesão carotídea, ateromatose aórtica, desfechos neurológicos e óbito. Resultados: Dentre 691 pacientes submetidos à CRM, 16 pacientes apresentaram acidentes vasculares cerebrais (AVC). Dentre esses, 11 pacientes (68.75%) apresentaram AVC localizados em áreas não compatíveis com as lesões carotídeas, sendo três deles com lesões calcificadas na aorta ascendente. Os pacientes com estenose carotídea apresentaram taxa similar de eventos neurológicos totais, AVC e óbito, comparados com pacientes sem estenose carotídea. Um subgrupo de 35 pacientes com estenose carotídea foi submetido à cirurgia coronariana com (14 pacientes) ou sem (21 pacientes) cirurgia de carótida, obtendo-se taxa de eventos neurológicos totais, AVC e óbito estatisticamente semelhantes. Os pacientes com calcificações aórticas apresentaram risco maior de eventos neurológicos (14,58% versus 6.55%, p=0.011), AVC (3,12% versus 2,18%, p=0,47) e óbito (8,33% versus 4,37%, p=0.12). Discussão: Os eventos neurológicos após CRM correlacionam-se com ateromatose aórtica. Os AVC freqüentemente não têm relação linear com a estenose carotídea. Estratégias para minimizar embolia da aorta podem diminuir as taxas de intercorrências neurológicas. / Introduction: The management of patients with simultaneous disease of carotid and coronary arteries is controversial. Studies showed that aortic calcifications might play a role in postoperative stroke at coronary artery bypass graft (CABG), carotid lesions may not be as important as previously considered. Method: A retrospective cohort of a randomly selected group (including elective and emergency operations) of 691 patients submitted to CABG was reviewed for general data, neurological complications and mortality. Results: Among 691 CABGs 16 patients had postoperative stroke. Among these, 11 patients (68.75%) had strokes not matching carotid lesions and anatomic presentation, three of those had detectable aortic calcifications. The patients with critical carotid stenosis had similar rates of neurological events, stroke and death as compared to patients without. The patients with aortic calcifications presented a higher risk of neurological events (14.58% versus 6.55%, p=0.011), stroke (3.12% versus 2.18%, p=0.47) and death (8.33% versus 4.37%, p=0.12). Discussion: The postoperative neurological events after CABG can be related to aortic calcifications. The strokes after coronary bypass may occur independently of the carotid lesions. Strategies to prevent aortic emboli may help preventing many post-operative strokes.

Objectives: Clinical implementation of 3D ultrasound (3D-US) in vascular surgery is in its infancy. The aim of this thesis was to develop novel clinical applications for 3D-US in the diagnosis and management of abdominal aortic aneurysm (AAA). Methods: Four principle clinical applications were investigated. 1) Intraoperative imaging – The ability of 3D-US to detect and classify endoleaks was compared with digital subtraction angiography in patients undergoing EVAR. 2) Detection and classification of endoleaks following endovascular aneurysm repair (EVAR) – The abilityof 3D-US to accurately detect and classify endoleaks following EVAR was compared to CTA and the final multi-disciplinary team decision. 3) AAA volume measurement – measurements using magnetic and optically-tracked 3D-US were compared to CTA. 4) Biomechanical analysis – the challenges of using 3D-US to generate surface models for biomechanical simulation was explored by development of an interactive segmentation technique and comparison of paired CT and 3D-US datasets. Optimal results were used in finite element analysis (FEA) and computational fluid dynamic(CFD) simulations. Results: 3D-US out-performed uniplanar angiography for the detection of endoleaks during EVAR. This approach allowed contrast-free EVAR to be performed in patients with poor renal function. 3D contrast-enhanced ultrasound was superior to CTA for endoleak detection and classification when compared with the final decision of the multi-disciplinary team. Optimal results for AAA volume measurements were gained using an optically tracked 3D-US system in EVAR surveillance. However, there remained a significant mean difference of 13.6ml between CT and 3D-US. Complete technical success of generating geometries for use in biomechanical analysis using 3D-US was only 5%. When the optimal results were used, a comparable CFD analysis under the conditions of steady, laminar and Newtonian flow was achieved. Using basic modelling assumptions in FEA, peak von Mises and principle wall stress was found to be at the same anatomical location on both the CT and 3D-US models but the 3D-US model overestimated the wall stress values by 41% and 51% respectively. Conclusions: 3D-US could be clinically implemented for intra-operative imaging and EVAR surveillance in specific cases. 3D-US volume measurement is feasible but future work should aim to improve accuracy and inter-observer reliability. Although the results of biomechanical analysis using the optimal results was encouraging and provided a proof-of-principal, there are a number of technical developments required to make this approach feasible in a larger number of patients.

Made available in DSpace on 2014-06-11T19:27:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-02-18Bitstream added on 2014-06-13T19:57:06Z : No. of bitstreams: 1 castro_m_me_jabo.pdf: 467319 bytes, checksum: 75c102107c67aefa122635ff9a85a085 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Aterosclerose é um condição inflamatória fibro-proliferativa crônica associada à produção de espécies oxidantes. O composto fenólico resveratrol encontrado principalmente na uva e no vinho tinto, parece ter atividades cardioprotetoras previnindo a oxidação de lipoproteínas de baixa densidade. Neste estudo investigou-se o efeito do resveratrol na prevenção da ateromatose induzida por meio de estudos morfológicos e morfométricos do arco aórtico, bem como observação do perfil bioquímico sérico de HDL, LDL, triglicerídeos e colesterol total. Para tanto, foram utilizados 20 coelhos divididos em quatro grupos os quais receberam as seguintes dietas durante 60 dias: grupo controle (CT) ração normal; grupo resveratrol (R) raçao normal e resveratrol na dose de 3 mg/Kg/dia; grupo colesterol (CL) ração acrescida de 1,5% de colesterol; grupo (CR) tratado com ração acrescida de 1,5% de colesterol e administração simultânea de resveratrol na dose de 3mg/Kg/dia. Na análise morfologica os animais do grupo CT e R não apresentaram alterações nas túnicas íntima, média e adventícia. Os animais do grupo CL apresentaram lesões ateroscleróticas com espessamento da íntima e invasão da média. Os animais do grupo CR também apresentaram lesões invadindo a íntima e média porém estas encontravam-se mais organizadas. O estudo morfométrico revelou maior espessamento da íntima no grupo CL, na camada média não foi observado diferença entre os grupos CL e CR. A avaliação do perfil lipídico dos animais dos grupos CL e CR revelou valores aumentados porém sem diferença entre estes grupos. Houve diferença quando comparados com os grupos que receberam uma dieta normal... / Atherosclerosis is a chronic fibro proliferative inflammatory disease associate to the production of reactive oxygen species. Resveratrol, a phenolic compound present in red wine and grape seems to prevent cardiovascular diseases by protecting low density lipoprotein from oxidation. The present study tested wheter resveratrol would provide any benefit in the prevention of induced atheroma lesions through morphologic, morphometric studies in the aortic arch and HDL, LDL, total cholesterol and triglyceride lipid levels. A total of 20 rabbits were divided into 4 groups during 60 days: group CT was given normal diet, group R normal diet with resveratrol at a dose of 3 mg/Kg/day, group CL normal diet supplemented with 1,5% cholesterol, group CR normal diet supplemented with 1,5% cholesterol with resveratrol at a dose of 3 mg/Kg/day. Morphologic analysis of the rabbit group CT and R didn’t show lesion in intima, media and adventitia tunicas in the aortic arch. Rabbits fed with a hypercholesterolemic diet show atherosclerotic lesions with thickness of intima and invasion of the media. Severity of atherosclerosis lesions was significantly reduced in group CR and lesions were more organized, and media was also affected. Morphometry study revealed that the intima of aortic arch in CL animals was thickner than CR animals. The media didn’t show difference between CL and CR groups. Lipid measurements of hypercholesterolemic rabbits showed a significant increase in serum levels but there weren’t differences between CL and CR group, there were differences when compared to those fed a regular diet...(Complete abstract, click electronic access below)

Made available in DSpace on 2014-06-11T19:23:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-07-23Bitstream added on 2014-06-13T18:51:20Z : No. of bitstreams: 1 castro_kf_me_jabo.pdf: 1173046 bytes, checksum: 3f548cfd5539d25d90bbd19c032374f5 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A aterosclerose é uma doença multifatorial, lenta e progressiva e a hiperlipidemia um dos fatores potenciais no desenvolvimento de doenças cardíacas ateroscleróticas. As vantagens da indução das dislipidemias experimentais são a produção de lesões ateromatosas em curto espaço de tempo; adequado controle dietético e fatores ambientais; a possibilidade de estudos sobre a reversibilidade de lesões ateroscleróticas e ensaios pré clínicos de substâncias hipolipidêmicas. Este estudo visou avaliar a capacidade do surfactante na indução aterogênica na região do arco aórtico de ratos por meio de estudos morfológico e morfométrico das túnicas íntima, média e adventícia da aorta, bem como analisar o perfil lipídico sérico. Foram utilizados 28 ratos Wistar, machos, albinos, adultos e hígidos. Estes foram distribuídos em quatro grupos experimentais formados por sete animais cada, a saber: Grupo I – (controle); Grupo II – tratado com tyloxapol, na dose de 500 mg/Kg de peso corporal, via intraperitoneal a cada 48 horas, durante duas semanas; Grupo III – tratado com tyloxapol na dose de 500 mg/Kg de peso corporal, via intraperitoneal a cada 48 horas, durante três semanas; Grupo IV – tratado com tyloxapol na dose de 500 mg/Kg de peso corporal, via intraperitoneal a cada 48 horas, durante quatro semanas. A análise morfológica do arco aórtico dos animais dos grupos II, III e IV evidenciou características histológicas semelhantes às do grupo I nas túnicas íntima, média e adventícia. No estudo morfométrico, os valores da espessura, em μm, da túnica íntima indicaram que o grupo III diferiu significativamente dos grupos I, II e IV. Os valores da espessura, em μm, encontrados para a túnica média e adventícia revelaram que não houve diferença significativa entre os grupos I, II, III e IV. Na avaliação do perfil lipídico os valores de colesterol total... / Atherosclerosis is a multifactorial, progressive and slow disease, and hyperlipidaemia is one of the potential factors in the development of atherosclerotic cardiac diseases. The experimental dyslipidaemia carrying out advantages are the production of atheromatous lesions in a short period of time, an adequate dietetic control and environmental factors, the possibility of studies concerning reversibility of atherosclerotic lesions, and pre-clinic experiments with hypolipidaemic substances. This study aims at evaluating tyloxapol ability in atherogenic inducing, in the rats’ aortic arch region, through morphologic morphometric studies, in intima, media and adventitia tunicas of the aortic arch, as well as analyzing serum lipid levels. Twenty-eight healthy Wistar adults’ albino male rats, weighing an average of 200 g were utilized. They were distributed into four experimental groups with seven animals each, as follows: Group I – (control); Group II – treated with tyloxapol at a dose of 500mg/Kg of body weight, through intraperitoneal via each 48 hours, for two weeks; Group III - treated with tyloxapol at a dose of 500mg/Kg of body weight, through intraperitoneal via each 48 hours, for three weeks; Group IV - treated with tyloxapol at a dose of 500mg/Kg of body weight, through intraperitoneal via each 48 hours, for four weeks. Morphological analysis of the aortic arch of the animals from groups II, III and IV, showed similar histological characteristics to group I at intima, media and adventitia tunicas. The intima tunica values at the morphometric study indicated that group III has significant differed from groups I, II and IV. The media and adventitia tunicas values revealed that there were no significant differences between groups I, II, III, and IV. As lipid profile evaluation is concerned, the values of total cholesterol, triglycerides and HDL have indicated... (Complete abstract click electronic access below)