Hyaluronan (HA) is a negatively charged extracellular matrix (ECM) component with water-attracting properties. It is the dominating ECM component in the renal medullary interstitium, where the amount changes in relation to hydration status: it increases during hydration and decreases during dehydration. It has, therefore, been suggested that HA participates in the regulation of renal fluid handling by changing the permeability properties of the interstitial space. This thesis investigates potential mechanisms for such a role in renal fluid regulation. The results demonstrate that the high renal HA content of late nephrogenesis decreases during the completion of kidney development in the rat, which takes place in the neonatal period. The heterogenous distribution of HA is mainly established during the first three weeks after birth. On day 21, the HA content is similar to that in the adult rat. The process is dependent on normal Ang II function. It primarily involves a reduction of HA synthase 2 expression and an increase of medullary hyaluronidase 1.  The cortical accumulation of HA that results from neonatal ACE inhibition can partly explain the pathological condition of the adult kidney, which causes reduced urinary concentration ability and tubulointerstitial inflammation. It is possible to reduce renomedullary HA with the HA synthesis inhibitor 4-MU, and the kidney’s ability to respond to a hydration challenge will then be suppressed, without affecting GFR.  The investigation of renomedullary interstitial cells (RMIC) in culture, shows that media osmolality and hormones of central importance for body fluid homeostasis, such as angiotensin II, ADH and endothelin, affect HA turnover through their effect on the RMICs, in a manner comparable to that found in vivo during changes in hydration status.  In established streptozotocin-induced diabetes, HA is regionally accumulated in the kidney, proteinuria and polyuria, reduced urine osmolality, and reduced response to ADH V2 activation will occur. As opposed to the proteinuria, the HA accumulation is not sensitive to mTOR inhibition, suggesting an alternate pathway compared to other ECM components  Taken together, the data suggest that during normal physiological conditions, renomedullary interstitial HA participates in renal fluid handling by affecting the interstitial prerequisites for fluid flux across the interstitial space. This is possible due to the water-attracting and physicochemical properties of this glycosaminoglycan. During pathological conditions, such as diabetes, the elevated interstitial HA can contribute to the defective kidney function, due to the proinflammatory and water-attracting properties of HA.

Approved for public release, distribution is unlimited / In order to address the requirements of the rapidly growing Internet, network processors have emerged as the solution to the customization and performance needs of networking systems. An important component in a network is the router, which receives incoming packets and directs them to specific routes elsewhere in the system. Network processors and the associated software control the routers and switches and allow software designers to deploy new systems such as multicasting forwarder and firewalls quickly.This thesis introduces network processors and their features, focusing on the Intel IXP1200 network processor. A multicast design for the IXP1200 using microACE is proposed. This thesis presents an approach to building a multicasting forwarder using the IXP1200 network processor layer-3 forwarder microACE that carries out unicast routing. The design is based on the Intel Internet exchange architecture and its active computing element (ACE). The layer-3 unicast forwarder microACE is used as a basic starting point for the design. Some software modules, called micoblocks, are modified to create a multicast forwarder that is flexible and efficient. / Lieutenant Junior Grade, Turkish Navy

Nous développons un modèle basé agents pour reproduire deux anomalies fréquemment observées sur les marchés financiers : distribution leptokurtique des rendements et ampleur de la volatilité irrégulière mais persistante de ces mêmes rendements. Notre but est de montrer de façon probante que ces anomalies pourraient être attribuées à une formation mimétique des anticipations des intervenants sur les marchés. Nous nous éloignons des développements récents dans le domaine des modèles modèles basés agents en finance pour proposer un modèle très simple, estimé à partir des traits statistiques saillants de l’indice français journalier CAC 40. L’hypothèse d’anticipations mimétiques peut ainsi être testée : elle n’est pas rejetée dans notre modélisation. / We develop an agent-based model to replicate two frequently observed anomalies in the financial markets: the fat tails and the clustered volatility of the distribution of the returns. Our goal is to show conclusively that these anomalies could be attributed to a mimetic formation of the expectations of the stakeholders in the markets. We did not follow the rencent developpments in the field of the ACE model in the finance, but we propose a very simple model which is estimated from the stylized facts of the French daily index CAC 40. The hypothesis of mimetic anticipations can thus be tested: it is not rejected in our modeling.

Interação entre angiotensina-(1-7) [Ang-(1-7)] e bradicinina (BK) foi determinada no mesentério de ratos Wistar anestesiados utilizando-se microscopia intravital. Aplicação tópica de BK e Ang-(1-7) induziram vasodilatação que foi abolida por HOE-140 e A-779, respectivamente. Ang-(1-7) (100 pmol) potencializou a vasodilatação de BK (1 pmol) mas não a vasodilatação promovida por acetilcolina, nitroprussiato de sódio, histamina e ácido araquidônico. O efeito potencializador de Ang-(1-7) sobre BK foi abolido por A-779, HOE-140, indometacina, L-NAME e TEA, entretanto, losartan não bloqueou este efeito. Enalaprilato aumentou a resposta vasodilatadora de BK e Ang-(1-7) e não alterou o efeito potencializador do segundo sobre o primeiro. Conclui-se que: 1)efeito potencializador de Ang-(1-7) sobre BK depende da interação de ambos com seus respectivos receptores; 2)é dependente de óxido nítrico, produtos da cicloxigenase e hiperpolarização de membrana via canais de potássio; 3) o mecanismo de potencialização parece não depender da atividade catalítica da ECA. / The interaction between angiotensin-(1-7) [Ang-(1-7)] and bradykinin (BK) was determined in the mesentery of anesthetized Wistar rats using intravital microscopy. The response-induced by topical application of BK (1, 10 and 30 pmol), Ang-(1-7) (1, 10, 100 and 1000 pmol) and Ang-(1-7) (100 pmol) + BK (1 pmol) was determined in mesenteric arterioles (15-20 mm diameter). The BK (1 pmol)- and Ang-(1-7) (100 pmol)- induced vasodilation was abolished by BK B2 receptor antagonist HOE-140 (100 pmol applied during 60 seconds) and the Ang-(1-7) antagonist [d-Ala7]-Ang-(1-7)] (A-779) (100 pmol applied during 15 seconds), respectively. Indomethacin (5 mg/kg; IM, 30 min before), a cyclooxygenase inhibitor; L-NAME (10 nmol; topical application, 3 min before), a NO synthase inhibitor, decreased the Ang-(1-7)-induced vasodilation. However, TEA (90 pmol; topical application), a non specific K+ channels blocker, did not alter the response to BK or Ang-(1-7). BK (1 pmol)-induced vasodilation, however, was potentiated by Ang-(1-7) 100 pmol. Sodium nitroprusside (38 pmol), acetylcholine (1,6 nmol), histamine (5,4 nmol) and arachdonic acid (10 nmol) responses were not modified by Ang-(1-7) 100 pmol. The Ang-(1-7)-potentiating effect on BK-induced vasodilation was abolished by A-779, HOE-140, indomethacin, L-NAME and TEA. Losartan (15 mg/kg.IV, 40 min before), an AT1 angiotensin receptor antagonist was without effect. On the other hand, enalaprilat treatment (10 mg/kg; IV, 30 min before), to inhibit angiotensin-converting enzyme (ACE), enhanced the BK and Ang-(1-7)-induced vasodilation but did not modify the effect of Ang-(1-7) on BK vasodilation. In conclusion, the potentiation of BK-induced vasodilation by Ang-(1-7) is a receptor-mediated phenomenon dependent on cyclooxygenase-related products, NO release and K+ channel-mediated membrane hyperpolarization. The potentiating mechanism, apparently, is not related to ACE catalytic activity.

O treinamento físico (TF) promove adaptações no sistema hematopoiético e o sítio NH2-terminal da enzima conversora de angiotensina I (ECA) hidrolisa um tetrapeptídeo hemorregulador negativo, o Acetil-Seril-Aspartil-Lisil-Prolina (Ac-SDKP). O objetivo do presente estudo foi investigar se o sítio NH2-terminal da ECA participaria nas adaptações hematopoiéticas induzidas pelo TF. Realizamos duas séries de experimentos. A primeira para determinarmos qual protocolo de TF seria o mais adequado para estudar as adaptações eritropoiéticas e a segunda para estudar o papel do sítio NH2-terminal da ECA nessas adaptações. Série de experimentos 1: ratas Wistar foram divididas em 3 grupos: controle (C), que realizaram TF (60 min/d, 5dias/sem) de 10 semanas uma vez ao dia (T1) e que realizaram o mesmo TF por 8 semanas, seguido de uma semana 2 vezes ao dia e uma semana 3 vezes ao dia (T2). Série de experimentos 2: ratas Wistar foram divididas em 4 grupos: controle (C), controle tratadas com captopril (10 mg.kg-1.dia-1) (C-Cap), treinadas sob o protocolo T2 (T2) e treinadas sob o protocolo T2 tratadas com captopril (T2-Cap). Foram medidos: 1) a pressão arterial (PA) e a freqüência cardíaca; 2) a hipertrofia cardíaca e atividade da citrato sintase; 3) o consumo máximo de oxigênio, o tempo de exercício e a distância percorrida em teste máximo; 4) a atividade catalítica dos terminais da ECA; 5) a concentração plasmática e na fração extracelular da medula óssea de Ac-SDKP; 6) o número e a proliferação de células tronco hematopoiéticas (CTH) no sangue e medula; 8) a reticulocitose e meia-vida das hemácias. As diferenças observadas apresentaram p<0,05. Série de experimentos 1: o protocolo T2 induziu maiores adaptações fisiológicas, morfológicas e funcionais em comparação ao T1. O protocolo T2 foi eficaz em promover adaptações no sistema eritropoiético como aumento no número e na capacidade proliferativa de CTH, no percentual de reticulócitos e redução na meia-vida das hemácias. O protocolo T2 aumentou a atividade catalítica do sítio NH2-terminal da ECA e reduziu a concentração plasmática e na fração extracelular da medula óssea de Ac-SDKP, o que não foi observado em T1. Série de experimentos 2: o grupo T2 apresentou aumento na atividade NH2-terminal da ECA, que foi inibido no grupo T2-Cap. A inibição do sítio NH2-terminal da ECA não influenciou a PA nem afetou as respostas ao treinamento. O grupo T2 apresentou redução na concentração plasmática e na fração extracelular da medula óssea de Ac-SDKP, enquanto no grupo T2-Cap não houve redução do Ac-SDKP no plasma e houve atenuação da redução na fração extracelular da medula óssea. Houve aumento no número e na proliferação de CTH na medula óssea e no sangue no grupo T2 e este aumento foi parcialmente inibido no grupo T2-Cap. Houve aumento na reticulocitose no grupo T2 e inibição parcial deste aumento no grupo T2-Cap. A meia-vida das hemácias foi reduzida em 50% no grupo T2, enquanto no grupo T2-Cap houve atenuação da redução. Concluímos que o protocolo de treino T2 estimula a hematopoiese pelo aumento na atividade do sítio NH2-terminal da ECA, aumento este que inativa o tetrapeptídeo Ac-SDKP. / Physical training (PT) promotes changes in the hematopoietic system and the NH2-terminal active site of angiotensin-converting enzyme I (ACE) hydrolyzes a tetrapeptide, negative hemoregulador, the acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP). The purpose of this study was to investigate whether the NH2-terminal active site of ACE plays a role in the hematopoietic changes induced by PT. We conducted two series of experiments. The first in prder to determine which PT protocol would be more appropriate to study the erythropoietic adaptations and the second to study the role of the NH2-terminal site of ACE in these adaptations. Experiment series 1: female Wistar rats were divided into 3 groups: control (C), submitted to PT (60 min/d, 5d/week) for 10 weeks once a day (T1) and submitted to the same TF for 8 weeks, followed by a week 2 times a day and a week 3 times a day (T2). Experiment series 2: female Wistar rats were divided into four groups: control (C), control treated with captopril (10 mg.kg-1.day-1) (C-Cap), trained under the protocol T2 (T2) and trained under protocol T2 treated with captopril (Cap-T2). We measured: 1) blood pressure (BP) and heart rate, 2) cardiac hypertrophy and citrate synthase activity, 3) the maximum oxygen consumption, exercise time and distance in maximal test, 4) catalytic activity ACE terminals, 5) plasma and bone marrow extracellular fraction concentration of Ac-SDKP, 6) the number and proliferation of hematopoietic stem cells (HSC) in the blood and marrow, 8) reticulocytosis and erythrocyte life-spam. The observed differences presented p <0.05. Experiment series 1: T2 protocol induced greater physiological, morphological and functional compared to T1. T2 protocol was effective in causing changes in the erythropoietic system such as increase the number and proliferative capacity of HSC, the percentage of reticulocytes and reduced erythrocyte life-spam. The protocol T2 increased the catalytic activity of the NH2-terminal site of ACE and decreased plasma and bone marrow extracellular fraction concentration of Ac-SDKP, which was not observed in T1. Experiment series 2: the T2 group showed an increase in the activity of the NH2-terminal ACE, which was inhibited in group T2-Cap. Inhibition of the NH2-terminal site of ACE did not influence or affect the BP responses to training. The T2 group showed a reduction in plasma and bone marrow extracellular fraction of Ac-SDKP, whereas in the T2-Cap there was no reduction of Ac-SDKP in plasma and there was attenuation of the reduction in the extracellular fraction of bone marrow. There was an increase in the number and proliferation of HSC in bone marrow and blood in the T2 group and this increase was partially inhibited in group T2-Cap. There was an increase in reticulocytosis in group T2 and partial inhibition of the increase in T2-Cap group. The erythrocyte life-spam was reduced by 50% in T2, while in the T2-Cap group there was attenuation of the reduction. We conclude that the training protocol T2 stimulates hematopoiesis by increasing the activity of the NH2-terminal site of ACE, an increase that inactivates the tetrapeptide Ac-SDKP.

O treinamento físico (TF) promove adaptações no sistema hematopoiético e o sítio NH2-terminal da enzima conversora de angiotensina I (ECA) hidrolisa um tetrapeptídeo hemorregulador negativo, o Acetil-Seril-Aspartil-Lisil-Prolina (Ac-SDKP). O objetivo do presente estudo foi investigar se o sítio NH2-terminal da ECA participaria nas adaptações hematopoiéticas induzidas pelo TF. Realizamos duas séries de experimentos. A primeira para determinarmos qual protocolo de TF seria o mais adequado para estudar as adaptações eritropoiéticas e a segunda para estudar o papel do sítio NH2-terminal da ECA nessas adaptações. Série de experimentos 1: ratas Wistar foram divididas em 3 grupos: controle (C), que realizaram TF (60 min/d, 5dias/sem) de 10 semanas uma vez ao dia (T1) e que realizaram o mesmo TF por 8 semanas, seguido de uma semana 2 vezes ao dia e uma semana 3 vezes ao dia (T2). Série de experimentos 2: ratas Wistar foram divididas em 4 grupos: controle (C), controle tratadas com captopril (10 mg.kg-1.dia-1) (C-Cap), treinadas sob o protocolo T2 (T2) e treinadas sob o protocolo T2 tratadas com captopril (T2-Cap). Foram medidos: 1) a pressão arterial (PA) e a freqüência cardíaca; 2) a hipertrofia cardíaca e atividade da citrato sintase; 3) o consumo máximo de oxigênio, o tempo de exercício e a distância percorrida em teste máximo; 4) a atividade catalítica dos terminais da ECA; 5) a concentração plasmática e na fração extracelular da medula óssea de Ac-SDKP; 6) o número e a proliferação de células tronco hematopoiéticas (CTH) no sangue e medula; 8) a reticulocitose e meia-vida das hemácias. As diferenças observadas apresentaram p<0,05. Série de experimentos 1: o protocolo T2 induziu maiores adaptações fisiológicas, morfológicas e funcionais em comparação ao T1. O protocolo T2 foi eficaz em promover adaptações no sistema eritropoiético como aumento no número e na capacidade proliferativa de CTH, no percentual de reticulócitos e redução na meia-vida das hemácias. O protocolo T2 aumentou a atividade catalítica do sítio NH2-terminal da ECA e reduziu a concentração plasmática e na fração extracelular da medula óssea de Ac-SDKP, o que não foi observado em T1. Série de experimentos 2: o grupo T2 apresentou aumento na atividade NH2-terminal da ECA, que foi inibido no grupo T2-Cap. A inibição do sítio NH2-terminal da ECA não influenciou a PA nem afetou as respostas ao treinamento. O grupo T2 apresentou redução na concentração plasmática e na fração extracelular da medula óssea de Ac-SDKP, enquanto no grupo T2-Cap não houve redução do Ac-SDKP no plasma e houve atenuação da redução na fração extracelular da medula óssea. Houve aumento no número e na proliferação de CTH na medula óssea e no sangue no grupo T2 e este aumento foi parcialmente inibido no grupo T2-Cap. Houve aumento na reticulocitose no grupo T2 e inibição parcial deste aumento no grupo T2-Cap. A meia-vida das hemácias foi reduzida em 50% no grupo T2, enquanto no grupo T2-Cap houve atenuação da redução. Concluímos que o protocolo de treino T2 estimula a hematopoiese pelo aumento na atividade do sítio NH2-terminal da ECA, aumento este que inativa o tetrapeptídeo Ac-SDKP. / Physical training (PT) promotes changes in the hematopoietic system and the NH2-terminal active site of angiotensin-converting enzyme I (ACE) hydrolyzes a tetrapeptide, negative hemoregulador, the acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP). The purpose of this study was to investigate whether the NH2-terminal active site of ACE plays a role in the hematopoietic changes induced by PT. We conducted two series of experiments. The first in prder to determine which PT protocol would be more appropriate to study the erythropoietic adaptations and the second to study the role of the NH2-terminal site of ACE in these adaptations. Experiment series 1: female Wistar rats were divided into 3 groups: control (C), submitted to PT (60 min/d, 5d/week) for 10 weeks once a day (T1) and submitted to the same TF for 8 weeks, followed by a week 2 times a day and a week 3 times a day (T2). Experiment series 2: female Wistar rats were divided into four groups: control (C), control treated with captopril (10 mg.kg-1.day-1) (C-Cap), trained under the protocol T2 (T2) and trained under protocol T2 treated with captopril (Cap-T2). We measured: 1) blood pressure (BP) and heart rate, 2) cardiac hypertrophy and citrate synthase activity, 3) the maximum oxygen consumption, exercise time and distance in maximal test, 4) catalytic activity ACE terminals, 5) plasma and bone marrow extracellular fraction concentration of Ac-SDKP, 6) the number and proliferation of hematopoietic stem cells (HSC) in the blood and marrow, 8) reticulocytosis and erythrocyte life-spam. The observed differences presented p <0.05. Experiment series 1: T2 protocol induced greater physiological, morphological and functional compared to T1. T2 protocol was effective in causing changes in the erythropoietic system such as increase the number and proliferative capacity of HSC, the percentage of reticulocytes and reduced erythrocyte life-spam. The protocol T2 increased the catalytic activity of the NH2-terminal site of ACE and decreased plasma and bone marrow extracellular fraction concentration of Ac-SDKP, which was not observed in T1. Experiment series 2: the T2 group showed an increase in the activity of the NH2-terminal ACE, which was inhibited in group T2-Cap. Inhibition of the NH2-terminal site of ACE did not influence or affect the BP responses to training. The T2 group showed a reduction in plasma and bone marrow extracellular fraction of Ac-SDKP, whereas in the T2-Cap there was no reduction of Ac-SDKP in plasma and there was attenuation of the reduction in the extracellular fraction of bone marrow. There was an increase in the number and proliferation of HSC in bone marrow and blood in the T2 group and this increase was partially inhibited in group T2-Cap. There was an increase in reticulocytosis in group T2 and partial inhibition of the increase in T2-Cap group. The erythrocyte life-spam was reduced by 50% in T2, while in the T2-Cap group there was attenuation of the reduction. We conclude that the training protocol T2 stimulates hematopoiesis by increasing the activity of the NH2-terminal site of ACE, an increase that inactivates the tetrapeptide Ac-SDKP.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A superativação do eixo ECA/AT1r está intimamente relacionada à síndrome metabólica e no organismo tem grande relação com o quadro de inflamação. A administração de frutose, seja por dieta ou pela água, tem sido usada como um modelo para a indução da superatividade desse eixo e para o estudo das vias inflamatórias relacionadas ao AT1r. Com isso, o objetivo deste trabalho foi avaliar se a administração de GW510156 poderia diminuir a superativação do eixo ECA/AT1r e consequentemente diminuir os danos causados pela dieta rica em frutose. Para isso foram utilizados camundongos machos C57Bl/6 que receberam uma dieta contendo 47% de frutose durante oito semanas ou uma dieta controle. Após oito semanas, os grupos foram redivididos aleatoriamente para o início da administração do GW501516 durante três semanas, totalizando quatro grupos experimentais. Os animais tratados apresentaram uma melhora da pressão arterial sistólica e também dos parâmetros urinários como proteinúria e ácido úrico. Houve ainda uma melhora dos triglicerídeo e ácido úrico plasmáticos. No tecido adiposo branco, o GW501516 foi capaz de diminuir a expressão dos componentes do eixo ECA/AT1r e também amenizou a inflamação causada pela dieta rica em frutose. No fígado, não houve alterações significativa do eixo, porém a fosforilação de JAK2 dependente de AT1r foi diminuída e consequentemente houve uma menor ativação das células estreladas no grupo que recebeu o GW501516. Além disso, as proteínas e genes relacionados à &#946;-oxidação foram aumentados com o tratamento e aqueles relacionados à lipogênese de novo, diminuídos o que resultou em menor esteatose no parênquima hepático. Os rins apresentaram uma melhora da inflamação induzida pelo eixo, apesar de o eixo também não ter apresentado diferenças significativas com o tratamento. Também não foram encontradas diferenças significativas na expressão proteica e gênica das proteínas antioxidantes. Com esses resultados podemos concluir que a curta administração do GW501516 pôde aliviar os efeitos inflamatórios e a esteatose hepática causada pela dieta rica em frutose, podendo ser pensado como uma nova ferramenta terapêutica no tratamento da superativação do eixo ECA/AT1r. / High-activation of ACE/AT1r axis is closely linked to metabolix syndrome and low-grade inflammation state. Fructose administration in water or in diet has been proposed as a model to study the high-activity of this axis and AT1r-related inflammatory pathways. In this view, we aimed to evaluate if GW501516 administration could decrease the high-activation of ACE/AT1r axis and consequently fructose damage. To this males mice C57Bl/6 were fed a high-fructose diet (47%) during eight weeks or standard-chow diet. After eight weeks, the groups were randomly divided to start treatment with GW501516, totaling four experimental groups. Animals treated with GW501516 presented an improvement of systolic blood pressure and in urinary parameters, as proteinuria and uric acid. Also was verified an improvement in plasmatic triglycerides and uric acid. In white adipose tissue, GW501516 was able to decrease the components of this axis and improved inflammation as well. In liver, it was not found differences in axis, but JAK2 phosphorylation AT1r-dependent was decreased and consequently it was found a diminished activations of hepatic stellate cells. In addition, proteins and genes related to &#946;-oxidation were increased with GW501516 and those related to lipogenesis de novo, were diminished, improving hepatic parenchyma. Kidneys presented an improvement of inflammation state, although it was not found differences in axis with treatment. Also, it was not found differences in gene and protein expression in relation to anti-oxidants proteins. These results show that short-administration of GW501516 could alleviate inflammatory effects and hepatic steatosis caused by high fructose diet, suggesting that GW501516 could be a new therapeutic option to treat the high activation of ACE/AT1r axis.

Peptídeos com ação inibitória da enzima conversora da angiotensina I (ECA) podem ser obtidos a partir de diversos alimentos e exercer efeito anti-hipertensivo. O cupuaçu (Theobroma grandiflorum S.), fruto nativo da Amazônia, possui sementes comestíveis contendo proteína de reserva similar à do cacau (Theobroma cacao L.), as quais parecem ser fonte de peptídeos inibidores da ECA. Desse modo, o objetivo deste trabalho foi investigar in vitro a ocorrência de peptídeos inibidores da ECA no hidrolisado proteico da semente de cupuaçu obtido por ação da Alcalase. O hidrolisado revelou o desaparecimento de proteínas entre 27 a 180 kDa, incluindo as globulinas, e o surgimento daquelas abaixo de 15 kDa após 2 h de hidrólise, indicando a formação de peptídeos menores. O ensaio de atividade utilizando o substrato Abz-FRK(Dnp)-P-OH revelou que o hidrolisado total promoveu 65% de inibição da ECA e esse pool peptídico foi fracionado em cinco frações (F1-F5) por cromatografia em fase reversa (RP-HPLC). Após a etapa de purificação, o monitoramento da inibição apontou, ao final, duas frações (3.2.8 e 3.4.10) com maior atividade inibitória. Oito peptídeos foram identificados por LC-MS/MS, sendo três deles já conhecidos como inibidores da ECA. Outros cinco novos peptídeos identificados (FLEK, GSGKHVSP, LDNK, MVVDKLF e MEKHS) foram sintetizados e tiveram sua ação inibitória validada por ensaios in vitro. O peptídeo GSGKHVSP apresentou a menor IC50 (3,11 &#181;M) e Ki (0,74 &#181;M), sendo um inibidor tipo misto quanto ao seu mecanismo de inibição revelado pelo gráfico de Lineweaver-Burk. Os resultados permitem concluir que o isolado proteico da semente de cupuaçu pode ser uma fonte para obtenção de peptídeos anti-hipertensivos, a despeito de serem necessárias investigações sobre a resistência desses peptídeos à digestão gastrointestinal e a eficácia da inibição in vivo. / Peptides with angiotensin I-converting enzyme (ACE) inhibitory activity may be obtained from several foods and cause antihypertensive effect. Cupuassu (Theobroma grandiflorum S.), a native fruit from Amazon, has edible seeds with a storage protein similar to that of cocoa (Theobroma cacao L.) which seems to have incrypted ACE inhibitor peptides. Thus, the aim of this project was to investigate the in vitro formation of ACE inhibitory peptides in protein hydrolysate from cupuassu seeds using Alcalase enzyme. The hydrolysate revealed the disappearance of proteins between 27 and 181 kDa after 2h hydrolysis, including the globulin, and the increase of those below 15 kDa, indicating the formation of peptides. The ACE inhibitory activity assays using the substrate Abz-FRK(Dnp)P-OH revealed the hydrolysate had 65% ACE inhibition and the pool of peptides was fractionated into five fractions (F1-F5) by reversed phase high-performance liquid chromatography (RP-HPLC). After the purification step, two fractions (3.2.8 e 3.4.10) exhibited the highest ACE-inhibitory activity. Eight peptides had been identified by LC-MS/MS and three of them were ACE inhibitors. The other newly identified peptides (FLEK, GSGKHVSP, LDNK, MVVDKLF and MEKHS) were synthesized and in vitro assayed for ACE inhibitory activity. The peptide GSGKHVSP had the lower IC50 (3.11 &#181;M) and Ki (0.74 &#181;M). Lineweaver-Burk plots suggest this peptide is a mixed-type inhibitor according to the inhibition mechanism. The results indicate that protein isolate from cupuassu seeds may be a good protein source of antihypertensive peptides and further investigation is needed in order to evaluate the resistance of these peptides to gastrointestinal digestion and the inhibitory activity in vivo.

Peptídeos com ação inibitória da enzima conversora da angiotensina I (ECA) podem ser obtidos a partir de diversos alimentos e exercer efeito anti-hipertensivo. O cupuaçu (Theobroma grandiflorum S.), fruto nativo da Amazônia, possui sementes comestíveis contendo proteína de reserva similar à do cacau (Theobroma cacao L.), as quais parecem ser fonte de peptídeos inibidores da ECA. Desse modo, o objetivo deste trabalho foi investigar in vitro a ocorrência de peptídeos inibidores da ECA no hidrolisado proteico da semente de cupuaçu obtido por ação da Alcalase. O hidrolisado revelou o desaparecimento de proteínas entre 27 a 180 kDa, incluindo as globulinas, e o surgimento daquelas abaixo de 15 kDa após 2 h de hidrólise, indicando a formação de peptídeos menores. O ensaio de atividade utilizando o substrato Abz-FRK(Dnp)-P-OH revelou que o hidrolisado total promoveu 65% de inibição da ECA e esse pool peptídico foi fracionado em cinco frações (F1-F5) por cromatografia em fase reversa (RP-HPLC). Após a etapa de purificação, o monitoramento da inibição apontou, ao final, duas frações (3.2.8 e 3.4.10) com maior atividade inibitória. Oito peptídeos foram identificados por LC-MS/MS, sendo três deles já conhecidos como inibidores da ECA. Outros cinco novos peptídeos identificados (FLEK, GSGKHVSP, LDNK, MVVDKLF e MEKHS) foram sintetizados e tiveram sua ação inibitória validada por ensaios in vitro. O peptídeo GSGKHVSP apresentou a menor IC50 (3,11 &#181;M) e Ki (0,74 &#181;M), sendo um inibidor tipo misto quanto ao seu mecanismo de inibição revelado pelo gráfico de Lineweaver-Burk. Os resultados permitem concluir que o isolado proteico da semente de cupuaçu pode ser uma fonte para obtenção de peptídeos anti-hipertensivos, a despeito de serem necessárias investigações sobre a resistência desses peptídeos à digestão gastrointestinal e a eficácia da inibição in vivo. / Peptides with angiotensin I-converting enzyme (ACE) inhibitory activity may be obtained from several foods and cause antihypertensive effect. Cupuassu (Theobroma grandiflorum S.), a native fruit from Amazon, has edible seeds with a storage protein similar to that of cocoa (Theobroma cacao L.) which seems to have incrypted ACE inhibitor peptides. Thus, the aim of this project was to investigate the in vitro formation of ACE inhibitory peptides in protein hydrolysate from cupuassu seeds using Alcalase enzyme. The hydrolysate revealed the disappearance of proteins between 27 and 181 kDa after 2h hydrolysis, including the globulin, and the increase of those below 15 kDa, indicating the formation of peptides. The ACE inhibitory activity assays using the substrate Abz-FRK(Dnp)P-OH revealed the hydrolysate had 65% ACE inhibition and the pool of peptides was fractionated into five fractions (F1-F5) by reversed phase high-performance liquid chromatography (RP-HPLC). After the purification step, two fractions (3.2.8 e 3.4.10) exhibited the highest ACE-inhibitory activity. Eight peptides had been identified by LC-MS/MS and three of them were ACE inhibitors. The other newly identified peptides (FLEK, GSGKHVSP, LDNK, MVVDKLF and MEKHS) were synthesized and in vitro assayed for ACE inhibitory activity. The peptide GSGKHVSP had the lower IC50 (3.11 &#181;M) and Ki (0.74 &#181;M). Lineweaver-Burk plots suggest this peptide is a mixed-type inhibitor according to the inhibition mechanism. The results indicate that protein isolate from cupuassu seeds may be a good protein source of antihypertensive peptides and further investigation is needed in order to evaluate the resistance of these peptides to gastrointestinal digestion and the inhibitory activity in vivo.

Distributed-memory machines offer a very high level of performance, flexibility and scalability. But the memory organization of this kind of machine determines that processes on different processors must communicate explicitly by sending and receiving messages. As a result, the programmer faces the enormously difficult task of detailed planning of algorithm-irrelevant, low-level communication issues. This level of programming resembles writing assembly programs for a sequential machine. ACE is a message-passing language with abstract communication statements. It was defined by Dr. Jingke Li at Portland State University. The communication in ACE is still explicit, but it is abstracted to a higher level. The abstraction can help balance the needs of ease of programming and high performance. This thesis discusses how those high-level communication abstractions can be transformed into low-level communication routines. It presents the design and implementation of a compiler that transforms an ACE program into a C program with low-level communication routines. The compiler is implemented for the Intel iPSC/2 hypercube multiprocessor machine. Compared to their low-level counterparts, ACE programs are easier to write and are more understandable. Compared to their high level counterparts, more efficient code can be generated since the communication information is expressed explicitly in ACE and the compiler itself is much less complex. ACE also enables the users to fine tune some critical communication segments. Some well known parallel algorithms written in ACE are compiled by the compiler as examples, and experimental results of their performance are included.