The present thesis shows the analysis, calibration, and application of the stress modified criticai strain criterion to predict ductile failure for an A285 steel. To obtain the mechanical behavior of the material, experimental tests were carried out by implementation of 5 different types of geometries: smooth round bar, notched round bar (R=1 , 2, 3 mm), and, deep and shallow cracked SE(B) specimens. Then, for the calibration process of the mechanical properties finite element models were generated, using 30 solid elements with 8 nodes (C308), matching the geometry and the properties of the tested specimens. To calibrate the elastoplastic behavior was used the experimental and numerical response obtained from the smooth and notched round bar specimens; and, for the damage calibration was used the responses obtained from both deep and shallow crack SE(B) specimens. Once the mechanical properties were calibrated, then there were obtained the SMSC criterion factors represented by the equation ..... and, the damage condition which is represented by the displacement at failure (.......) and exponential factor (....). This calibrated model was able to recover the SE(B) experimental responses that validate the use of the characterized material in a complex structure. Then, the fully characterized material was applied in two pipelines which have externai initial circumferential elliptical crack; being the first one pipe with shallow crack and the second one with deep crack. Finally, both pipes were submitted to tension loads to predict the ductile damage behavior, obtaining the necessary load to the crack start growing, and the evolution of the failure. / A presente dissertação apresenta o processo de análise, calibração e aplicação das propriedades mecânicas, incluindo o comportamento elastoplástico e de dano, para o aço A285, utilizando o critério \"Stress modified criticai strain\" (SMCS). Para obter o comportamento mecânico do material, testes experimentais foram realizados com a implementação de 5 tipos diferentes de geometrias: barra cilíndrica sem entalhe, barra cilíndrica com entalhe (R = 1, 2, 3 mm) e corpos de prova SE(B) com trinca inicial profunda e rasa. Para o processo de calibração das propriedades mecânicas foram gerados modelos de elementos finitos, utilizando elementos sólidos 30 com 8 nós (C3D8), que representam de forma adequada a geometria e as propriedades dos corpos de prova testados. Para calibrar o comportamento elastoplástico e iniciação do dano, utilizou-se a resposta experimental e numérica obtida para as amostras de barra cilíndrica com e sem entalhe; e, para a calibração da evolução do dano, foram utilizadas as respostas obtidas para os espécimes SEB de trincas profundas e rasa. Este modelo calibrado foi capaz de recuperar as respostas experimentais dos corpos de prova SE(B), o que valida o uso do material caracterizado em uma estrutura complexa. Uma vez calibradas as propriedades mecânicas, foram obtidos os fatores do critério SMSC representados pela equação ....... , e, a condição de dano que é representada pelo deslocamento na falha .... e o fator de amolecimento exponencial .... . Depois, o material totalmente caracterizado foi aplicado em dois dutos que possuem trinca elíptica circunferencial inicial externa; sendo o primeiro tubo com trinca superficial e o segundo com trinca profunda. Finalmente, ambos os tubos foram submetidos a cargas de tensão para prever o comportamento do dano dúctil, obtendo a carga necessária para o início do crescimento da trinca e a evolução da falha.

The present thesis shows the analysis, calibration, and application of the stress modified criticai strain criterion to predict ductile failure for an A285 steel. To obtain the mechanical behavior of the material, experimental tests were carried out by implementation of 5 different types of geometries: smooth round bar, notched round bar (R=1 , 2, 3 mm), and, deep and shallow cracked SE(B) specimens. Then, for the calibration process of the mechanical properties finite element models were generated, using 30 solid elements with 8 nodes (C308), matching the geometry and the properties of the tested specimens. To calibrate the elastoplastic behavior was used the experimental and numerical response obtained from the smooth and notched round bar specimens; and, for the damage calibration was used the responses obtained from both deep and shallow crack SE(B) specimens. Once the mechanical properties were calibrated, then there were obtained the SMSC criterion factors represented by the equation ..... and, the damage condition which is represented by the displacement at failure (.......) and exponential factor (....). This calibrated model was able to recover the SE(B) experimental responses that validate the use of the characterized material in a complex structure. Then, the fully characterized material was applied in two pipelines which have externai initial circumferential elliptical crack; being the first one pipe with shallow crack and the second one with deep crack. Finally, both pipes were submitted to tension loads to predict the ductile damage behavior, obtaining the necessary load to the crack start growing, and the evolution of the failure. / A presente dissertação apresenta o processo de análise, calibração e aplicação das propriedades mecânicas, incluindo o comportamento elastoplástico e de dano, para o aço A285, utilizando o critério \"Stress modified criticai strain\" (SMCS). Para obter o comportamento mecânico do material, testes experimentais foram realizados com a implementação de 5 tipos diferentes de geometrias: barra cilíndrica sem entalhe, barra cilíndrica com entalhe (R = 1, 2, 3 mm) e corpos de prova SE(B) com trinca inicial profunda e rasa. Para o processo de calibração das propriedades mecânicas foram gerados modelos de elementos finitos, utilizando elementos sólidos 30 com 8 nós (C3D8), que representam de forma adequada a geometria e as propriedades dos corpos de prova testados. Para calibrar o comportamento elastoplástico e iniciação do dano, utilizou-se a resposta experimental e numérica obtida para as amostras de barra cilíndrica com e sem entalhe; e, para a calibração da evolução do dano, foram utilizadas as respostas obtidas para os espécimes SEB de trincas profundas e rasa. Este modelo calibrado foi capaz de recuperar as respostas experimentais dos corpos de prova SE(B), o que valida o uso do material caracterizado em uma estrutura complexa. Uma vez calibradas as propriedades mecânicas, foram obtidos os fatores do critério SMSC representados pela equação ....... , e, a condição de dano que é representada pelo deslocamento na falha .... e o fator de amolecimento exponencial .... . Depois, o material totalmente caracterizado foi aplicado em dois dutos que possuem trinca elíptica circunferencial inicial externa; sendo o primeiro tubo com trinca superficial e o segundo com trinca profunda. Finalmente, ambos os tubos foram submetidos a cargas de tensão para prever o comportamento do dano dúctil, obtendo a carga necessária para o início do crescimento da trinca e a evolução da falha.

During atherosclerosis and restenosis, there is an extensive amount of collagen synthesis and degradation. Changes in the types of collagen present can have profound effects on vascular smooth muscle cell (SMC) proliferation and migration. Type VIII collagen, which is normally present at low levels within the mature vascular system, is greatly increased during atherogenesis. The central theme of this thesis is to determine the role of type VIII collagen in the pathogenesis of atherosclerosis and restenosis. In the first study, we demonstrated the importance of type VIII collagen in SMC migration and proliferation. SMCs from type VIII collagen-deficient mice display increased adhesion and decreased spreading, migration, and proliferation compared to SMCs from wild-type mice. Treatment of SMCs from type VIII collagen-deficient mice with exogenous type VIII collagen can rescue the defects. In the second study, we determined that type VIII collagen exerts its effects through regulation of MMP-2 expression. Type VIII collagen-deficient SMCs have decreased levels of MMP-2 and are impaired in chemotaxis toward PDGF-BB and in their ability to contract thick collagen gels. We found that decreasing endogenous MMP-2 levels in normal SMCs or adding exogenous collagen to type VIII collagen-deficient SMCs is sufficient to recapitulate the type VIII collagen-deficient or wild-type SMC phenotype, respectively. In the third study, we investigated the contribution of type VIII collagen to intimal hyperplasia after mechanical injury in the mouse. We found that type VIII collagen-deficient mice display a 35% reduction in intimal hyperplasia and attenuated vessel remodeling after femoral artery wire injury, establishing a role for type VIII collagen in restenosis. The results of the work presented in this thesis demonstrate that production of type VIII collagen confers an SMC phenotype with a greater propencity for proliferation and migration. These effects are in part mediated through regulation of MMP-2 expression and activation. We conclude that the increases in type VIII collagen production that occur during atherosclerosis and restenosis contribute to the capacity of SMCs to alter the existing extracellular matrix in a manner which permits enhanced migration.

During atherosclerosis and restenosis, there is an extensive amount of collagen synthesis and degradation. Changes in the types of collagen present can have profound effects on vascular smooth muscle cell (SMC) proliferation and migration. Type VIII collagen, which is normally present at low levels within the mature vascular system, is greatly increased during atherogenesis. The central theme of this thesis is to determine the role of type VIII collagen in the pathogenesis of atherosclerosis and restenosis. In the first study, we demonstrated the importance of type VIII collagen in SMC migration and proliferation. SMCs from type VIII collagen-deficient mice display increased adhesion and decreased spreading, migration, and proliferation compared to SMCs from wild-type mice. Treatment of SMCs from type VIII collagen-deficient mice with exogenous type VIII collagen can rescue the defects. In the second study, we determined that type VIII collagen exerts its effects through regulation of MMP-2 expression. Type VIII collagen-deficient SMCs have decreased levels of MMP-2 and are impaired in chemotaxis toward PDGF-BB and in their ability to contract thick collagen gels. We found that decreasing endogenous MMP-2 levels in normal SMCs or adding exogenous collagen to type VIII collagen-deficient SMCs is sufficient to recapitulate the type VIII collagen-deficient or wild-type SMC phenotype, respectively. In the third study, we investigated the contribution of type VIII collagen to intimal hyperplasia after mechanical injury in the mouse. We found that type VIII collagen-deficient mice display a 35% reduction in intimal hyperplasia and attenuated vessel remodeling after femoral artery wire injury, establishing a role for type VIII collagen in restenosis. The results of the work presented in this thesis demonstrate that production of type VIII collagen confers an SMC phenotype with a greater propencity for proliferation and migration. These effects are in part mediated through regulation of MMP-2 expression and activation. We conclude that the increases in type VIII collagen production that occur during atherosclerosis and restenosis contribute to the capacity of SMCs to alter the existing extracellular matrix in a manner which permits enhanced migration.

Platelet-derived growth factor D-chain (PDGF-D) is the newest member of the PDGF family of mitogens and chemo-attractants; it is expressed in a wide variety of cell types, including vascular smooth muscle cells (SMCs). The molecular mechanisms regulating PDGF-D transcription are unknown. Here I investigated the effects of angiotensin II (ATIl) and IL-1 beta on the transcription of PDGF-D and changes in vascular SMCs phenotype. Primer extension analysis mapped a single transcriptional start site to the ccAG CGC motif of PDGF-D promoter. Several potential transcription factor binding sites such as SpI, Ets-1, NF-??B, IRF-1, p53, Smad4 and AP1 were located in the proximal 1168bp of the PDGF-D promoter. ATII-inducible Ets-1 and PDGF-D gene expression is mediated via H202. IL-I beta supresses PDGF-D promoter activity, mRNA and protein expression in SMCs through NF-??B p65, IRF-1 and HDAC1, which form complex in the PDGF-D promoter. This study provides the first direct link between NF-KB and the PDGF-D promoter, IRF-1 with any member of the PDGF family and a new example of HDAC mediated inhibition of gene expression. In summary, this study investigates for the first time the mechanisms mediating the transcriptional regulation of PDGF-D in vascular SMCs. This provides valuable insights into the molecular control of vascular phenotype, and opens up potential opportunities for therapeutic intervention.

Hållbar utveckling är ett aktuellt ämne i dagens samhälle och det ställs allt högre krav på att företag ska ta sitt ansvar för att bidra till en hållbar utveckling. Bankerna är de aktörer som kan styra hur kapital ska investeras och har därmed stor makt och mycket att vinna på att främja en hållbar utveckling i samhället. De globala affärsmöjligheterna som marknaden för hållbar utveckling utgör uppskattas att år 2050 uppgå till omkring tre biljoner dollar per år. Banking on shared value (BSV), som är en vidareutveckling av creating shared value, innebär att banker kan generera ekonomiskt värde på ett sätt som samtidigt skapar socialt och miljömässigt värde. Av en granskning som har utförts av Fair Finance Guide framgår det att de sju största bankerna i Sverige får relativt låga hållbarhetsbetyg. Många forskare menar att sustainability management control systems (SMCS), vilket är en en utvecklad modell av Malmi och Browns teori om MCS kan vara nyckeln till att lyckas integrera hållbar utveckling i verksamheten. Det efterfrågas dock fler praktikfall inom området. Syftet med denna studie är att bidra till en ökad förståelse för hur företagsledningen inom banksektorn kan använda SMCS för att integrera hållbar utveckling i sin verksamhet En fallstudie har genomförts av Ekobanken som är den enda svenska bank som är medlem i Global Alliance for Banking on Values och som har fått högst betyg av samtliga banker i Fair Finance Guides granskning. Det empiriska materialet har samlats in med hjälp av intervjuer med två personer inom Ekobankens ledning samt från dokumentstudier. Studien har en abduktiv ansats. Vi har identifierat Ekobankens mest framträdande styrningsverktyg som kulturell styrning som har ett starkt fokus på hållbar utveckling, administrativ styrning och cybernetiska styrningsverktyg. Ekobanken har enligt oss ett väl fungerande SMCS som lyckas med att integrera hållbar utveckling i verksamheten. Vi anser dock inte att Ekobankens arbete med hållbar utveckling kan definieras som BSV fullt ut men att de är den bank i Sverige som i nuläget kommer närmast att uppfylla kraven för BSV. Vi menar därmed att studiens resultat med fördel kan verka inspirerande för andra banker. Det är framförallt Ekobankens transparens, öppna klimat, ledningens engagemang samt deras hållbarhetsredovisning och utlåningspolicy som andra banker, som jobbar mot hållbar utveckling, enligt vår mening bör ta fasta på. / Sustainable development is a topical subject in today's society and there is an increasing demand for companies to take responsibility by contributing to sustainable development. The banks control how capital is invested, thus giving them great power to and a lot to gain from promoting sustainable development in society. The business opportunities from the market for sustainable development is estimated to provide a value of three trillion dollars annually by 2050. Banking on shared value ​​(BSV), which is a further development of creating shared value, means that banks can generate economic value and at the same time create social and environmental value. A review conducted by Fair Finance Guide shows that the seven largest banks in Sweden are performing relatively low results in terms of sustainability. Many researchers argue that sustainability management control systems (SMCS), which is a developed model of Malmi and Brown's theory about MCS, can be the key to successfully integrating sustainable development into the business. However, more practical cases are required in the area. This study aims at contributing to an increased understanding of how corporate governance in the banking sector can use SMCS to integrate sustainable development into the business. A case study has been carried out on Ekobanken, which is the only Swedish bank that is a member of Global Alliance for Banking on Values and has been awarded the highest rating in the Fair Finance Guide´s review. The empirical material has been collected by means of semi-structured interviews with two persons within Ekobanken´s management team as well as documentary studies. The study has an abductive approach. We have identified Ekobanken's most prominent management tools as cultural governance with a strong focus on sustainable development, administrative governance and cybernetic management tools. In our opinion Ekobanken has a successful SMCS that manage to integrating sustainable development into the business. However, Ekobanken's work with sustainable development can, in our opinion, not be fully defined as BSV, but we believe that they are the bank in Sweden that is currently closest to meeting the requirements for BSV. Therefore we mean that the results of this study can be of inspiration for other banks. It is primarily Ekobanken's transparency, open climate, management's commitment, and their sustainability report and lending policy that other banks working towards sustainable development should strive to apply.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal. / L'anion superoxyde peut agir comme une molécule de signalisation ou comme un facteur préjudiciable selon sa concentration, l'organe cible, et selon la présence ou non d'antioxydants neutralisants. Actuellement, dans les cellules musculaires lisses (CMLs) vasculaires, les effets de l'anion superoxyde sur les différentes voies de transduction du signal et sur les interactions croisées entre ces voies ne sont pas encore définies. Par conséquent, une meilleure connaissance des effets de l'anion superoxyde sur les différentes voies de signalisation pourrait fournir une meilleure compréhension des mécanismes sous-jacents aux fonctions altérées des CMLs vasculaires observées dans des conditions pathologiques. L'objectif général de cette étude était de caractériser et d'évaluer le rôle modulateur de l'anion superoxyde, produit par la réaction de l'hypoxanthine avec la xanthine oxidase, sur les activités de différentes voies de signalisation dans les CMLs vasculaires, et de déterminer si la sensibilité de différentes voies de signalisation à l'anion superoxyde était altérée dans l'hypertension artérielle. Le projet de ce programme de recherche était basé sur les principaux postulats suivants : (1) l'anion superoxyde pourrait affecter sélectivement la production d'inositol 1,4,5-triphosphates (IP3), de GMPc, ou d'AMPc dans les CMLs vasculaires; (2) le rôle modulateur de l'anion superoxyde pourrait être dû à une altération des interactions croisées entre différentes voies de signalisation; et (3) les anomalies observées dans les CMLs vasculaires chez le rat spontanément hypertendu (SHR) pourraient être reliées à des altérations des différentes voies de signalisation induites par l'anion superoxyde. Une production augmentée d'1P3induite par l'anion superoxyde dans les CMLs d'aorte de rat ou d'artère mésentérique en culture a été démontrée pour la première fois dans cette étude. L'anion superoxyde a augmenté la formation d'IP3d'une manière concentration-dépendante et temps-dépendante. La superoxyde dismutase (SOD), mais non la catalase, a inhibé significativement la formation d'IP3 induite par l'anion superoxyde. L'inhibition de la phospholipase C (PLC) a aboli l'effet de l'anion superoxyde sur la formation d'1P3. La génistéine et la tyrphostine A25, deux inhibiteurs de la tyrosine kinase, ont aussi inhibé significativement la formation d'IP3induite par l'anion superoxyde. L'utilisation d'anticorps anti-PLCy a atténué significativement la formation d'1P3induite par l'anion superoxyde. De plus, le taux d'expression des protéines de la PLCy a été augmenté après l'exposition des CMLs à l'anion superoxyde. Ces observations suggèrent donc que dans les CMLs vasculaires la formation d'1P3 induite par l'anion superoxyde pourrait être en grande partie secondaire à une augmentation de l'activité de la tyrosine kinase liée aux voies de signalisation de la PLCy. En ce qui concerne la voie du GMPc, l'anion superoxyde a diminué significativement les niveaux de base de GMPc et supprimé aussi l'augmentation des niveaux de GMPc induite par des stimulateurs de la guanylyl cyclase, le nitroprussiate de sodium (NPS) ou la s-nitroso-nacétylpénicillamine (SNAP). La formation d'1P3stimulée par l'anion superoxyde a été significativement inhibée par le NPS ou la SNAP, mais potentialisée de façon importante par un inhibiteur de la guanylyl cyclase l'ODQ ou par le KT5823 (un inhibiteur de la protéine kinase dépendant du GMPc). Cependant, l'anion superoxyde n'a pas eu d'effet sur les niveaux de base d'AMPc ou sur la production d'AMPc induite par la forskoline et de plus, l'inhibition de l'adénylyl cyclase ou de la protéine kinase dépendante de l'AMPe n'a pas affecté la formation d'lP3stimulée par l'anion superoxyde. Ces données, par conséquent, suggèrent que l'inhibition de la formation de GMPc par l'anion superoxyde contribue probablement à l'activation de la formation d'1P3induite par l'anion superoxyde en atténuant le rétrocontrôle inhibiteur du GMPc sur les voies de signalisation liées à la PLC, tandis que la voie de signalisation de l'AMPc ne serait pas impliquée dans la formation d'EP3induite par l'anion superoxyde. Dans les CMLs vasculaires de rat SHR, les effets de l'anion superoxyde ont été plus puissants que dans les CMLs de rat WKY, en ce qui concerne l'augmentation de formation d'1P3, la diminution des taux de GMPc et la facilitation induite par l'anion superoxyde des interactions croisées entre les voies du GMPc et de 1'IP3. Dans les CMLs vasculaires des deux souches de rat, la formation d'IP3induite par l'anion superoxyde a été inhibée par une variété d'antioxydants, dont la N-acétylcystéine, l'acide a-lipoïque, la mélatonine et la SOD. Il apparaît donc vraisemblable que l'hypersensibilité à l'anion superoxyde des voies de 1'IP3et du GMPc puissent contribuer à l'augmentation du tonus vasculaire et de la réactivité des CMLs dans l'hypertension artérielle. Nous avons aussi investigué si l'effet de la mélatonine était dû à ses propriétés antioxydantes. Un effet inhibiteur plus important de la mélatonine sur la contraction aortique induite par la norépinéphrine (NE) a été observé chez les rats SHR en comparaison avec les rats Wistar-Kyoto (WKY). L'inhibition par la mélatonine de la formation d'IP induite par la NE a été aussi plus importante dans les CMLs aortiques de rat SHR que dans celles de rat WKY. Les effets plus puissants de la mélatonine chez le rat SHR, qui ont été aussi observés avec la SOD, mais non avec la catalase, ne sont pas dûs à l'activation des récepteurs à la mélatonine ou des récepteurs a-adrénergiques. Ces résultats indiquent que les effets anti-hypertenseurs de la mélatonine sont largement dûs à l'inactivation de l'anion superoxyde, et que les niveaux endogènes d' antioxydants ne parviennent pas à contrecarrer les niveaux accrus d'anion superoxyde produits chez le rat SHR. En conclusion, cette étude révèle une variété de nouveaux mécanismes de signalisation de l'anion superoxyde. Pour la première fois, il a été démontré que l'anion superoxyde active l'hydrolyse des phosphoinositides et augmente les taux d'IP3dans les CMLs vasculaires, principalement par la stimulation de la tyrosine kinase liée à la voie de signalisation de la PLCy. Il a aussi été observé que l'anion superoxyde réduit la formation de GMPc et supprime l'inhibition croisée de 1'1P3par le GMPc, facilitant ainsi la formation d'1P3. Les effets sélectifs de l'anion superoxyde sur les voies de 1'IP3et du GMPc, ainsi que l'existence d'une inhibition croisée de la formation d'1P3par la voie du GMPc, révèlent des mécanismes nouveaux pour expliquer le rôle modulateur de l'anion superoxyde sur les voies de signalisation dans les CMLs. Par conséquent, les effets plus puissants de l'anion superoxyde sur la signalisation de la voie de 1'IP3et de la voie du GMPc dans les CMLs vasculaires de rat SHR, effets qui ont été démontrés pour la première fois dans cette étude, pourraient être responsables des altérations des mécanismes de transduction du signal cellulaire chez le rat SHR et ainsi contribuer au développement et/ou au maintien de l'hypertension artérielle. Ces observations permettent donc d'imaginer de nouvelles orientations pour le développement de nouvelles stratégies pour la prévention ou le traitement de l'hypertension artérielle. / Superoxide anion can act as a signalling molecule or a detrimental factor depending on its concentration, the targeted organ, and the presence of counteracting antioxidants. The effects of superoxide on different signal transduction pathways and on the cross-talk interactions among these pathways in vascular smooth muscle cells (SMCs) are presently still unsettled. Therefore, a better knowledge on the effects of superoxide on different signalling pathways may provide a better understanding of the mechanisms underlying the altered functions in vascular SMCs observed in pathological conditions. The general objective of this study was to characterize and evaluate the modulating role of superoxide generated by the hypoxanthine and xanthine oxidase reaction on the activities of different signalling pathways in vascular SMCs and to investigate whether the sensitivities of different signalling pathways to superoxide were altered in hypertension. The design of the present research program was based on the following major postulates. (1) superoxide might selectively affect the generation of inositol 1,4,5-triphosphates (IP3), cGMP, or cAMP in vascular SMCs; (2) the modulating role of superoxide might be mediated by alteration in the cross-talk interactions among different signalling pathways; and (3) the abnormalities observed in vascular SMCs from spontaneously hypertensive rats (SHR) might be related to the alterations induced by superoxide on different signalling pathways. An enhanced production of 1P3induced by superoxide in cultured SMCs from rat aorta or mesenteric artery was demonstrated, for the first time, in this study. Superoxide increased 1P3 formation in a concentration- and time-dependent manner. Superoxide dismutase (SOD), but not catalase, significantly inhibited the superoxide-increased 1P3formation. The inhibition of phospholipase C (PLC) abolished the effect of superoxide on IP3formation. Genistein and tyrphostin A25, two tyrosine kinase inhibitors, also significantly inhibited the superoxideinduced IP3formation. The application of antibody against PLCI, significantly attenuated the superoxide-induced 1P3formation. Moreover, the expression level of PLC7proteins was increased after exposing SMCs to superoxide. These observations thus suggest that superoxideinduced IP3 formation may be in a great part secondary to an increase in the activity of tyrosine kinase-link PLCy signalling pathways in vascular SMCs. Concerning the cGMP pathway, superoxide significantly decreased the basal levels of cGMP and also suppressed the rise in cGMP levels induced by guanylyl cyclase stimulator sodium nitroprusside (SNP) or s-nitroso-n-acetylpenicillamine (SNAP). The superoxide-induced IP3 formation was significantly inhibited by SNP or SNAP, but markedly potentiated by a guanylyl cyclase inhibitor ODQ or KT5823 (a cGMP-dependent protein kinase inhibitor). However, superoxide had no effect on the basal levels of cAMP or the forskolin-induced cAMP production and moreover, the inhibition of adenylyl cyclase or cAMP-dependent protein kinase did not affect the superoxide-enhanced IP3formation. These data, therefore, suggest that the reduced cGMP formation by superoxide probably contributes to the superoxide induced activation of 1P3 formation by lifting the inhibitory feedback of cGMP on the PLC pathway(s), whereas, the cAMP pathway may not be involved in the superoxide-induced IP3formation. In vascular SMCs from SHR, the effects of superoxide were more potent than in SMCs from WKY, including the increase in 1P3 formation, the decrease in cGMP levels, and the superoxide-induced facilitation of the cross-talk interaction between cGMP and IP3pathways. The superoxide-induced 1P3formation was inhibited by a variety of antioxidants, including nacetylcysteine, cc-lipoic acid, melatonin and SOD, in vascular SMCs from both strains. It thus appears that the hypersensitivity of 1P3and cGMP pathways to superoxide is likely to contribute to the increased vascular tone and reactivity of SMCs in hypertension. Whether the effect of melatonin is due to its antioxidant properties was also explored. A greater inhibitory effect of melatonin on the norepinephrine (NE)-induced aortic contraction was observed in SHR than in Wistar-Kyoto rats (WKY). The inhibition of the NE-induced IP formation by melatonin was also greater in aortic SMCs from SHR than that from WKY. The enhanced effects of melatonin in SHR, which were found to be similarly enhanced with SOD but not with catalase, were not mediated by melatonin receptors or oc-adrenoceptors. These results indicate that the anti-hypertensive effects of melatonin are largely due to the scavenging of superoxide, and that the levels of endogenous antioxidants may not counteract the levels of overproduced superoxide in SHR. In conclusion, this study reveals a variety of novel signalling mechanisms for superoxide. For the first time, it was demonstrated that superoxide activates the hydrolysis of phosphoinositides and increases IP3levels in vascular SMCs mainly through the stimulation of tyrosine kinase-link PLCy signal pathway. It was also found that superoxide reduces cGMP formation and suppresses the cross-inhibition of IP3by cGMP, thus facilitating 1133formation. The selective effects of superoxide on 1133and cGMP pathways as well as the existence of a cross-inhibition of IP3formation by cGMP pathway provide novel mechanisms for the signalling role of superoxide in vascular SMCs. Therefore, the altered signalling effects of superoxide on the IP3pathway and the cGMP pathway, which were demonstrated in vascular SMCs from SHR for the first time in this study, could thus be responsible for the alterations in cellular signal transduction mechanisms in SHR and might contribute to the development and/or maintenance of hypertension. These observations could provide new avenues for the development of new strategies for the prevention or treatment of hypertension.