O objetivo do presente trabalho foi analisar o papel do ANP na ação não genômica da Aldosterona sobre o trocador Na+/H+ no segmento S3 do túbulo proximal de rato, isolado, in vitro. Os resultados indicam que o pHi basal do segmento S3 proximal de ratos é 7.20 + ou - 0.009 (n = 47/209). O valor médio da velocidade de extrusão celular de H+ na condição controle é de 0.195 + ou - 0.012 pHi/min (n = 16/96). Os dados confirmam que a aldosterona apresenta um efeito bifásico sobre o NHE1: em baixas doses (10-12 M) o estimula, enquanto que em altas doses (10-6M), o inibe. O ANP (10-6 M) não possui efeito sobre o NHE1; contudo, o ANP previne ambos os efeitos da aldosterona sobre esse trocador. O valor médio da concentração do cálcio no citosol ([Ca2+]i) na condição controle é 100 ± 1 (n = 5) hM Adicionalmente, nossos estudos mostram que o ANP diminui a [Ca2+]i e inibe o efeito estimulatório de ambas as doses de aldosterona sobre esse parâmetro. / The effects of aldosterone and ANP(2 min preincubation) on the intracellular pH recovery rate (pHirr) after the acid load induced by NH4Cl and on the [Ca2+]i were investigated in isolated rat S3 segment. The basal pHi was 7.20 + ou - 0.009(n=47/209) and the basal pHirr via the Na+/H+ exchanger was 0.195 + ou - 0.012 pHi/min(n=16/96). Aldosterone(10-12M) caused an increase in the pHirr, but aldosterone(10-6M) decreased it. ANP(10-6M) alone or plus aldosterone(10-12 or 10-6 M) had no effect on pHirr. The basal [Ca2+]i was 100 + ou - 1(n=5)hM. After 1 min of Aldosterone pi there was a transient and dose-dependent increase of the [Ca2+]i and after 6 min pi there was a new increase of [Ca2+]i. ANP alone decreased the [Ca2+]i and prevented the stimulatory effects of aldosterone(10-12 or 10-6M) on this parameter. The data indicate a nongenomic action of aldosterone and ANP on the Na+/H+ exchanger and on [Ca2+]i and are compatible with stimulation of the this exchanger by increases in [Ca2+]i in the lower range (at10-12M aldosterone) and inhibition by increases at high levels (at10-6M aldosterone).

Submitted by Mariana Tsuha Moreno (mariana.moreno@raizen.com) on 2016-10-21T13:48:46Z No. of bitstreams: 1 Dissertação (v 2016 - v final).pdf: 1034853 bytes, checksum: 5c83d9a498dbbf7d7c57a487e570299d (MD5) / Approved for entry into archive by Joana Martorini (joana.martorini@fgv.br) on 2016-10-21T13:52:47Z (GMT) No. of bitstreams: 1 Dissertação (v 2016 - v final).pdf: 1034853 bytes, checksum: 5c83d9a498dbbf7d7c57a487e570299d (MD5) / Made available in DSpace on 2016-10-21T14:45:46Z (GMT). No. of bitstreams: 1 Dissertação (v 2016 - v final).pdf: 1034853 bytes, checksum: 5c83d9a498dbbf7d7c57a487e570299d (MD5) Previous issue date: 2016-09-02 / Essential condition for the proper functioning of the new State model sought by the Federal Constitution of 88, the Brazilian regulatory system is significantly behind the American congeneric in what concern to the employment of advanced regulatory practices. Taking the national petroleum agency-ANP as a reference, not much besides public consultations and hearings are required for the issuing of new regulation. Tools such as (1) mechanisms for coordination, planning and control of the regulatory action, (2) the fixing of a minimum content for the regulatory reasons, including the obligatory usage of regulatory impact analysis, and (3) the compulsory periodic review of existing regulation, among several other tools incorporated by the American system throughout the evolution experimented in the 20th century, are whether not used or not mandatory. Such fact, combined with an unreasoned tendency to deference by the Judiciary when asked to review such agent’s action, explains how a crucial agency like ANP remains indifferent to such practices after two decades since its foundation. Given that the conclusions of this work are based on a single case of a single agency, such conclusions cannot (and should not) be taken as a representative sample of the modus operandi of all agencies or, in general, as a representative sample of the quality of the entire Brazilian regulatory system. And this is not, in fact, the objective; our claim is that due to the absence of a regulatory framework requiring the adoption of uniform and mandatory regulatory practices, combined with the undemanding attitude demonstrated by the Judiciary in the control of the actions of these agents, the Brazilian regulatory system admits such a low level of performance that should be cause for concern and immediate attention. / Condição fundamental para o bom funcionamento do novo modelo de Estado pretendido pela Constituição Federal de 88, o sistema regulatório brasileiro está significativamente atrasado em relação ao seu congênere norte-americano no que se refere ao emprego de boas práticas de regulação. Tomando-se como referência a Agência Nacional do Petróleo – ANP, veremos neste trabalho que não se encontram nas normas de criação das agências brasileiras muito mais do que menções genéricas a necessidade de realização de consultas e audiências públicas para a discussão de propostas de regulação que, de alguma maneira, possam afetar direitos dos entes regulados. Ferramentas como (1) mecanismos de coordenação, planejamento e controle da ação regulatória, (2) fixação de conteúdo mínimo para a fundamentação da ação regulatória pretendida, incluindo a exigência da apresentação de análise de impacto regulatório e detalhamento sobre seu decision-making process, e (3) a obrigatoriedade de revisão periódica da regulação existente, dentre várias outras ferramentas incorporadas ao sistema americano ao longo da evolução pela qual este sistema passou ao longo da segunda metade do século XX, ou não são utilizadas ou não são obrigatórias. Tal fato, aliado à tendência verificada à deferência não fundamentada que o Judiciário demonstra quando chamado a realizar o controle judicial das normas editadas por estes agentes, é o que permite que uma agência tão importante quanto ANP, quase duas décadas após a sua criação, permaneça indiferente à tais práticas. Evidentemente, por estarem as conclusões deste trabalho baseadas na análise da experiência concreta de um único caso, de uma única agência, tais conclusões não podem (e não devem) ser tomadas como amostra representativa do modus operandi de todas as agências ou, de maneira geral, da qualidade de todo o sistema regulatório brasileiro. E não é esta, de fato, a pretensão; o argumento que se pretende marcar é que, em razão da ausência de um marco regulatório que obrigue a adoção uniforme e obrigatória de boas práticas de regulação, combinada com a postura pouco exigente demonstrada pelo Judiciário no controle dos atos destes agentes, o sistema regulatório brasileiro, em seu estágio atual de evolução, admite atuação como a que veremos no caso concreto da ANP, o que deve ser motivo de preocupação e atenção imediata.

O objetivo do presente trabalho foi analisar o papel do ANP na ação não genômica da Aldosterona sobre o trocador Na+/H+ no segmento S3 do túbulo proximal de rato, isolado, in vitro. Os resultados indicam que o pHi basal do segmento S3 proximal de ratos é 7.20 + ou - 0.009 (n = 47/209). O valor médio da velocidade de extrusão celular de H+ na condição controle é de 0.195 + ou - 0.012 pHi/min (n = 16/96). Os dados confirmam que a aldosterona apresenta um efeito bifásico sobre o NHE1: em baixas doses (10-12 M) o estimula, enquanto que em altas doses (10-6M), o inibe. O ANP (10-6 M) não possui efeito sobre o NHE1; contudo, o ANP previne ambos os efeitos da aldosterona sobre esse trocador. O valor médio da concentração do cálcio no citosol ([Ca2+]i) na condição controle é 100 ± 1 (n = 5) hM Adicionalmente, nossos estudos mostram que o ANP diminui a [Ca2+]i e inibe o efeito estimulatório de ambas as doses de aldosterona sobre esse parâmetro. / The effects of aldosterone and ANP(2 min preincubation) on the intracellular pH recovery rate (pHirr) after the acid load induced by NH4Cl and on the [Ca2+]i were investigated in isolated rat S3 segment. The basal pHi was 7.20 + ou - 0.009(n=47/209) and the basal pHirr via the Na+/H+ exchanger was 0.195 + ou - 0.012 pHi/min(n=16/96). Aldosterone(10-12M) caused an increase in the pHirr, but aldosterone(10-6M) decreased it. ANP(10-6M) alone or plus aldosterone(10-12 or 10-6 M) had no effect on pHirr. The basal [Ca2+]i was 100 + ou - 1(n=5)hM. After 1 min of Aldosterone pi there was a transient and dose-dependent increase of the [Ca2+]i and after 6 min pi there was a new increase of [Ca2+]i. ANP alone decreased the [Ca2+]i and prevented the stimulatory effects of aldosterone(10-12 or 10-6M) on this parameter. The data indicate a nongenomic action of aldosterone and ANP on the Na+/H+ exchanger and on [Ca2+]i and are compatible with stimulation of the this exchanger by increases in [Ca2+]i in the lower range (at10-12M aldosterone) and inhibition by increases at high levels (at10-6M aldosterone).

Bradykinin (BK) acts through eNOS to reduce MAPK-mediated feedback inhibition of insulin signalling. Preliminary data suggest that the sGC-cGMP-PKG pathway, a prominent NO target, is involved. Our present study aimed to support the role of this pathway with atrial natriuretic peptide (ANP), which uses a receptor associated GC (NPR-A) to generate cGMP. We found that treating adipocytes with ANP mimicked BK effects on insulin-stimulated glucose uptake, Tyr-IRS-1 and Akt/PKB phosphorylation, as well as JNK and ERK1/2 inhibition. These outcomes depended on GC-cGMP-PKG signalling since A71915 (NPR-A antagonist), and KT-5823 (PKG inhibitor), completely abrogated them, while zaprinast (phosphodiesterase inhibitor), prolonged ANP actions. Furthermore, decreased MAPK phosphorylation was independent of upstream kinase activity, suggesting that MAPK phosphatases may be involved. These data indicate that BK and ANP act through the GC-cGMP-PKG pathway to potentiate insulin signalling via attenuated feedback inhibition. Stimulating the GC-cGMP-PKG pathway may, therefore, be a promising therapy for T2DM.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Core enterprise is the center of Green Supply Chain (GSC) and it helps partners to meet the environmental standards and to improve GSC’s performance. However there are two questions for core enterprise when it establishes GSC: What roles do core enterprise play in GSC? How does core enterprise improve GSC’s performance? What criterions are considered when core enterprise chooses partners in GSC?   In order to answer these questions, this paper will use following methods: firstly, Analytic hierarchy process (AHP) and Analytic network process (ANP) have be used to study on three research companies (IKEA, TOYOTA and Apple); Secondly, six interviews with research companies’ employees.   After case study, the results show that when core enterprise chooses partners, it will draw up some criterions and sub-standards for partners to fulfill. Furthermore, core enterprise plays the roles as GSC’s builder, director, coordinator, and designer in the system.   Finally, this paper answered the research questions: Firstly, in order to improve the performance of GSC, core enterprise should direct all partners in GSC to save materials and recycle waste in each link of GSC. Secondly, core enterprise can use four criterions to choose partners. These criterions are: environmental management performance, product-related elements, production-service levels, and enterprise culture.

Adopting new technologies enable enterprises to improve employees¡¦ performance and competitive advantages. The differences in natures of adopting processes of organizations and individuals need further clarify for better understandings regarding to their framework in adopting new technologies. This paper based on the Theory of Reasoned Action (TRA) and existed research to construct the relations amongst the effective factors which affect the adopting new technologies in either individual or organization perspectives. In the individual level, the constructed research framework indicated employees¡¦ e-Learning satisfaction could be measured by three major dimensions, the perceived e-Learning qualities, individual internal beliefs (usefulness and ease of use), and social influence. Eight proposed hypothesis were confirmed by Structured Equation Modeling analysis of 428 valid samples. Path analyses verified the original path in TRA, TAM, and D&M ISS Model. The perceived e-Learning qualities and social influence cause significantly influence to employees¡¦ e-Learning satisfaction in both directly and indirectly, which by way of individual internal beliefs, positive paths. The results also showed that perceived information quality, usefulness, system quality, social influence, ease of use, and than service quality positively affect employees¡¦ satisfaction of e-Learning in descend sequences. Where, in the organization level, decision framework of adopting new technology of oil refinery was composed by modified Delphi method and was verified by Analytic Network Process from the survey of 15 experts. The consistency opinions confirmed four inter-depended dimensions and seventeen criteria were included. The results suggested that process fitness, environmental fitness, actors¡¦ organizational fitness, and new technology characteristics are important dimensions of adopting new technology in descend sequences. On the other hand, economic feasibility, relative advantages, government, environment acceptance, and engineering feasibility are the top five important factors to be evaluated during the adopting process. The different natures of adopting processes of organizations and individuals cause their different framework in adopting new technologies. This paper concluded that new technology, actors¡¦, environmental characteristics are three interdepended dimensions which influence the adopting behavior no matter in individual or organization context. In organization level of oil refinery case, actors¡¦ characteristics consist not only of actors¡¦ organizational fitness but also process fitness, which is the most important dimension while adopting new technology. In final, the implications of findings were discussed and directions were also suggested for future research.

H οικογένεια των γουανυλικών κυκλασών περιλαμβάνει δύο μέλη, μία διαλυτή μορφή (sGC) και μια μορφή που είναι συνδεδεμένη στην κυτταρική μεμβράνη (pGC). Αυτές ενεργοποιούνται από τα διαφορετικούς προσδέτες. Η μεν sGC από το μονοξείδιο του αζώτου (NO), η δε pGC από τα νατριουρητικά πεπτίδια και με την ενεργοποίησή τους παράγουν την κυκλική GMP (cGMP). Σε πολλές περιπτώσεις, η ξεχωριστή αυτή παραγωγή του cGMP από sGC έναντι αυτής που προέρχεται από την pGC οδηγεί σε διαφορετικές βιολογικές δράσεις. Προηγούμενη εργασία στο εργαστήριό μας έχει δώσει έμφαση στη σημασία της από sGC παραγόμενης cGMP στην αγγειογένεση. Ωστόσο, περιορισμένες πληροφορίες είναι διαθέσιμες όσον αφορά στις αγγειογενετικές δράσεις της pGC στο ενδοθήλιο. Επομένως επιδιώξαμε να καθορίσουμε τα αποτελέσματα της ενεργοποίησης pGC στις ιδιότητες των ενδοθηλιακών κυττάρων που σχετίζονται με την αγγειογένεση. Αρχικά, ερευνήσαμε την επίδραση του κολπικού νατριουρητικού πεπτιδίου (ANP) στο σχηματισμό αγγείων αίματος in vivo, χρησιμοποιώντας ως πρότυπο μοντέλο τη χοριοαλλαντοϊκή μεμβράνη εμβρύου όρνιθας (CAM). Το ANP (0.1-10 μmole) ενίσχυσε τη νεοαγγειογένεση με δοσοεξαρτώμενο τρόπο, όπως διαπιστώθηκε από την αύξηση στα σημείων διακλάδωσης και το μήκος αγγείων. In vitro, το ANP ενίσχυσε τον πολλαπλασιασμό (0.01-1 μΜ) και τη μετανάστευση (10 μΜ) ενδοθηλιακών κυττάρων ανθρώπινου ομφάλιου λώρου (HUVEC) και υποκίνησε το σχηματισμό δομών αγγειακού τύπου από τα HUVEC 10 μΜ) σε υπόστρωμα Matrigel. Προκειμένου να μελετηθούν οι μηχανισμοί που ενέχονται στην επαγόμενη από ANP αγγειογένεση, εξετάσαμε τη φωσφορυλίωση δύο κινασών (MAPK), των ERK1/2 και p38, ως πιθανά μόρια-στόχουςστο μονοπάτι της σηματοδότησης του ANP. Το ANP ενεργοποίησε τόσο την ERK1/2, όσο και την p38 MAPKs κατά χρονοεξαρτώμενο τρόπο. Για να εξασφαλίσουμε στοιχεία για τη λειτουργική σημασία p38 χρησιμοποιήσαμε ένα φαρμακολογικό της αναστολέα, τον παράγοντα SB203580. Προεπώαση των κυττάρων με SB203580 ανέστειλε εν μέρει τη μετανάστευση των HUVEC. Εν περιλήψει, τα στοιχεία μας δείχνουν ότι το ANP προάγει την αγγειογένεση in vivo και ενισχύει τις σχετιζόμενες με αγγειογένεση, ιδιότητες των ενδοθηλιακών κυττάρω in vitro με τη ρύθμιση της φωσφορυλίωσης των MAPK. Η ενεργοποίηση λοιπόν της pGC να είναι ευεργετική όταν απαιτείται ο σχηματισμός νέων αγγείων αίματος. / The guanylyl cyclase (GC) family comprises of two members, a soluble one (sGC) and a membrane-bound GC, the particulate GC (pGC). These are activated by different ligands; sGC by nitric oxide (NO) and pGC by natriuretic peptides and upon activation produce cyclic GMP (cGMP). In many instances compartmentalized production of cGMP by sGC vs pGC results in different biological responses. Previous work in our laboratory has highlighted the importance of sGC-derived cGMP in angiogenesis. However, limited information is available with regard to the angiogenic actions of pGC in the endothelium. We therefore sought to determine the effects of pGC activation on angiogenesis-related properties of EC. Initially, we investigated the effects of atrial natriuretic peptide (ANP) on blood vessel formation in vivo, using as a model the chick embryo chorioallantoic membrane (CAM). ANP (0.1-10 μmole) enhanced neovascularization in a dose-dependent manner, as shown by the increase in branching points and vessel length. In vitro, ANP increased human umbilical vein endothelial cell (HUVEC) growth (0.01-1 μΜ) and migration (10 μΜ) and stimulated the assembly of HUVEC into tube-like networks (10 μΜ) on Matrigel. In order to study the mechanisms implicated in ANP-induced angiogenesis, we examined the phosphorylation of two MAP Kinases (MAPK), ERK1/2 and p38, as possible downstream targets of ANP signaling. ANP activated both ERK1/2 and p38 MAPKs in a time-dependent manner. To provide evidence for the functional significance of p38 we used the pharmacological inhibitor SB203580. Pretreatment of cells with SB203580 inhibited ANF-stimulated migration of HUVEC. In summary, our data show that ANP promotes angiogenesis in vivo and enhances angiogenesis-related properties of endothelial cells in vitro by modulating phosphorylation of MAPK. pGC activation might be beneficial when new blood vessel formation is desired.

Bradykinin (BK) acts through eNOS to reduce MAPK-mediated feedback inhibition of insulin signalling. Preliminary data suggest that the sGC-cGMP-PKG pathway, a prominent NO target, is involved. Our present study aimed to support the role of this pathway with atrial natriuretic peptide (ANP), which uses a receptor associated GC (NPR-A) to generate cGMP. We found that treating adipocytes with ANP mimicked BK effects on insulin-stimulated glucose uptake, Tyr-IRS-1 and Akt/PKB phosphorylation, as well as JNK and ERK1/2 inhibition. These outcomes depended on GC-cGMP-PKG signalling since A71915 (NPR-A antagonist), and KT-5823 (PKG inhibitor), completely abrogated them, while zaprinast (phosphodiesterase inhibitor), prolonged ANP actions. Furthermore, decreased MAPK phosphorylation was independent of upstream kinase activity, suggesting that MAPK phosphatases may be involved. These data indicate that BK and ANP act through the GC-cGMP-PKG pathway to potentiate insulin signalling via attenuated feedback inhibition. Stimulating the GC-cGMP-PKG pathway may, therefore, be a promising therapy for T2DM.