An ANP model to support decision-making in a Portuguese pharmaceutical supply chain

Dissertação para obtenção do Grau de Mestre em Engenharia e Gestão Industrial

Correlations between ANP concentrations in atria, plasma and cerebral structures and sodium chloride preference in Wistar rats

We determined whether ANP (atrial natriuretic peptide) concentrations, measured by radioimmunoassay, in the ANPergic cerebral regions involved in regulation of sodium intake and excretion and pituitary gland correlated with differences in sodium preference among 40 Wistar male rats (180-220 g). Sodium preference was measured as mean spontaneous ingestion of 1.5% NaCl solution during a test period of 12 days. The relevant tissues included the olfactory bulb (OB), the posterior and anterior lobes of the pituitary gland (PP and AP, respectively), the median eminence (ME), the medial basal hypothalamus (MBH), and the region anteroventral to the third ventricle (AV3V). We also measured ANP content in the right (RA) and left atrium (LA) and plasma. The concentrations of ANP in the OB and the AP were correlated with sodium ingestion during the preceding 24 h, since an increase of ANP in these structures was associated with a reduced ingestion and vice-versa (OB: r = -0.3649, P<0.05; AP: r = -0.3291, P<0.05). Moreover, the AP exhibited a correlation between ANP concentration and mean NaCl intake (r = -0.4165, P<0.05), but this was not the case for the OB (r = 0.2422). This suggests that differences in sodium preference among individual male rats can be related to variations of AP ANP level. Earlier studies indicated that the OB is involved in the control of NaCl ingestion. Our data suggest that the OB ANP level may play a role mainly in day-to-day variations of sodium ingestion in the individual rat

Does plasma ANP participate in natriuresis induced by a-MSH?

a-Melanocyte-stimulating hormone (a-MSH; 0.6 and 3 nmol) microinjected into the anteroventral region of the third ventricle (AV3V) induced a significant increase in diuresis without modifying natriuresis or kaliuresis. Intraperitoneal (ip) injection of a-MSH (3 and 9.6 nmol) induced a significant increase in urinary sodium, potassium and water excretion. Intraperitoneal (3 and 4.8 nmol) or iv (3 and 9.6 nmol) administration of a-MSH did not induce any significant changes in plasma atrial natriuretic peptide (ANP), suggesting that the natriuresis, kaliuresis and diuresis induced by the systemic action of a-MSH can be dissociated from the increase in plasma ANP. These preliminary results suggest that a-MSH may be involved in a g-MSH-independent mechanism of regulation of hydromineral metabolism

Interactions of ANP and ANG II in tubular nephron acidification

(ANP, 1 &micro;M) on the kinetics of bicarbonate reabsorption in the rat middle proximal tubule, we performed in vivo experiments using a stopped-flow microperfusion technique with the determination of lumen pH by Sb microelectrodes. These studies confirmed that ANG II added to the luminal or peritubular capillary perfusion fluid stimulates proximal bicarbonate reabsorption and showed that ANP alone does not affect this process, but impairs the stimulation caused by ANG II. We also studied the effects and the interaction of these hormones in cortical distal nephron acidification. Bicarbonate reabsorption was evaluated by the acidification kinetic technique in early (ED) and late (LD) distal tubules in rats during in vivo stopped-flow microperfusion experiments. The intratubular pH was measured with a double-barreled microelectrode with H+-sensitive resin. The results indicate that ANG II acted by stimulating Na+/H+ exchange in ED (81%) and LD (54%) segments via activation of AT1 receptors, as well as vacuolar H+-ATPase in LD segments (33%). ANP did not affect bicarbonate reabsorption in either segment and, as opposed to what was seen in the proximal tubule, did not impair the stimulation caused by ANG II. To investigate the mechanism of action of these hormones in more detail, we studied cell pH dependence on ANG II and ANP in MDCK cells using the fluorescent probe BCECF. We showed that the velocity of cell pH recovery was almost abolished in the absence of Na+, indicating that it is dependent on Na+/H+ exchange. ANP (1 &micro;M) alone had no effect on this recovery but reversed both the acceleration of H+ extrusion at low ANG II levels (1 pM and 1 nM), and inhibition of H+ extrusion at higher ANG II levels (100 nM). To obtain more information on the mechanism of interaction of these hormones, we also studied their effects on the regulation of intracellular free calcium concentration, [Ca2+]i, monitored with the fluorescent probe Fura-2 in MDCK cells in suspension. The data indicate that the addition of increasing concentrations of ANG II (1 pM to 1 &micro;M) to the cell suspension led to a progressive increase in [Ca2+]i to 2-3 times the basal level. In contrast, the addition of ANP (1 &micro;M) to the cell suspension led to a very rapid 60% decrease in [Ca2+]i and reduced the increase elicited by ANG II, thus modulating the effect of ANG II on [Ca2+]i. These results may indicate a role of [Ca2+]i in the regulation of the H+ extrusion process mediated by Na+/H+ exchange and stimulated/impaired by ANG II. The data are compatible with stimulation of Na+/H+ exchange by increases of [Ca2+]i in the lower range, and inhibition at high [Ca2+]i levels

Relationship between the actions of atrial natriuretic peptide (ANP), guanylin and uroguanylin on the isolated kidney

Guanylin and uroguanylin are peptides that bind to and activate guanylate cyclase C and control salt and water transport in many epithelia in vertebrates, mimicking the action of several heat-stable bacteria enterotoxins. In the kidney, both of them have well-documented natriuretic and kaliuretic effects. Since atrial natriuretic peptide (ANP) also has a natriuretic effect mediated by cGMP, experiments were designed in the isolated perfused rat kidney to identify possible synergisms between ANP, guanylin and uroguanylin. Inulin was added to the perfusate and glomerular filtration rate (GFR) was determined at 10-min intervals. Sodium was also determined. Electrolyte dynamics were measured by the clearance formula. Guanylin (0.5 µg/ml, N = 12) or uroguanylin (0.5 µg/ml, N = 9) was added to the system after 30 min of perfusion with ANP (0.1 ng/ml). The data were compared at 30-min intervals to a control (N = 12) perfused with modified Krebs-Hanseleit solution and to experiments using guanylin and uroguanylin at the same dose (0.5 µg/ml). After previous introduction of ANP in the system, guanylin promoted a reduction in fractional sodium transport (%TNa+, P<0.05) (from 78.46 ± 0.86 to 64.62 ± 1.92, 120 min). In contrast, ANP blocked uroguanylin-induced increase in urine flow (from 0.21 ± 0.01 to 0.15 ± 0.007 ml g-1 min-1, 120 min, P<0.05) and the reduction in fractional sodium transport (from 72.04 ± 0.86 to 85.19 ± 1.48, %TNa+, at 120 min of perfusion, P<0.05). Thus, the synergism between ANP + guanylin and the antagonism between ANP + uroguanylin indicate the existence of different subtypes of receptors mediating the renal actions of guanylins.

Le syndrome métabolique chez les congéniques du rat Dahl : influence de la diète et rôle du récepteur de l'ANP

L’hypertension artérielle et l’obésité sont deux composantes conjointement reliées du syndrome métabolique. Les récepteurs de l’ANP (GCA) et de l’oxyde nitrique (GCs) ont des propriétés diurétiques, natriurétiques, vasodilatatrices et sont liés au contrôle de la pression. Des études récentes ont démontré leur implication dans l’obésité. Hypothèse : Une différence génétique au niveau du gène GCA pourrait contribuer à des différences physiologiques. La composante lipidique et/ou sodique de la diète pourrait influencer la fonction rénale, cardiaque et les valeurs anthropométriques différemment chez les souches congéniques. Objectifs : (1) Déterminer l’effet de la composante lipidique et sodique des diètes; (2) Évaluer l’influence de GCA sur la réponse physiologique des souches congéniques; (3) Expliquer les mécanismes physiologiques procurant une réduction de la pression artérielle chez la souche SM9. Méthodologie : Des modèles congéniques du rat Dahl (DSS) hypertendu, nourri avec une diète riche en gras (HF) ou normale (NF), ont été utilisés pour démontrer l’impact d’un segment chromosomique d’origine normotendue. Résultats : La souche SM9 a une prise de poids plus importante que SM12 et DSS sur diète HF malgré un apport alimentaire équivalent. La souche SM9 présente également un ratio masse adipeuse/masse maigre plus élevé que SM12 et DSS. Nous n’avons observé aucune augmentation de la pression artérielle en réponse à la diète HF pour les 3 souches malgré une augmentation du dommage rénal pour les 3 souches. Le dommage rénal est plus important chez DSS que pour les 2 congéniques. La réponse diurétique à l’ANP est plus élevée chez SM9 et est influencée par le contenu en sel dand la diète. La perte glomérulaire plus importante chez le rat DSS semble compensée par une augmentation de la réponse à l’ANP par les glomérules résiduels. Il y a une corrélation entre l’activité de GCA en réponse à l’ANP, les niveaux d’ARNm et le nombre de répétition du dinucléotide TA dans son promoteur. Le rat DSS présente une hypertrophie cardiaque plus importante que les deux souches congénique et ceci n’est pas modifié par la diète HF. Conclusion : Nos études ont permis de mettre en évidence un effet génétique impliquant le segment chromosomique normotendu contenant GCA dans la réponse à une diète HF chez le rat DSS.

Qualidade de vida no serviço público: um programa para a Agência Nacional do Petróleo, Gás Natural e Biocombustíveis - ANP

This study aimed to verify which are the main factors for a Quality of Working Life Program in context of the Agência Nacional do Petróleo, Gás Natural e Biocombustíveis (ANP). For this, a descriptive, an explanatory, a literature, a documentary and a field research was applied. ANP was the universe of the research. The sample consists of 2 Quality of Working Life Program¿s managers: one of Agência Nacional de Águas (ANA), and another one from Agência Nacional de Saúde Suplementar (ANS), as well as managers and servers from ANP. Two semi structured interviews, a mixed questionnaire and a words evoke test were applied. The Quality of Working Life Factors used in this study were: working conditions, health, moral, compensation, participation, internal communication, organization image, headmansubaltern relationship and working organization. It was concluded that relevant factors for a Quality of Working Life Program in ANP are: working organization, working condition, moral, health and organization image.

Desafios que enfrenta a ANP para regular a indústria de petróleo no Brasil depois da quebra do monopólio em 1997 ate o ano de 2005

This work is analyzing the challenges which the National Petrol Agency is facing to regulate the Petrol industry in Brazil after the Monopoly crash in the period between 1997 until 2005. Due to the necessities of adaptation of its political strategies to the rules which determine the international economic flows, Brazil was forced to use the Economic Regulation in order to control the market. The regulation established in Brazil is not indifferent to imperfect markets. Thus can be find a conflict of interests among companies, the government and consumers within this process of regulation. The established agency does not have enough autonomy for administrating a regulation. The State with its paternalism power does not allow the agency to fulfill its function for which it was established, even though its function was established by law. A regulating policy which is clearly defined will establish a strong and independent agency with a clear limitation of its competences, avoiding divergent interpretation which prioritizes investments and promotes economic development. The agency will have the challenge to regulate the companies that enter the sector, allowing the opening of the market for new initiatives of investments which contribute to the welfare of the country and breaking at the same time the monopoly that is lead by Petrobras since 1953. Combining a stable set of rules with agility in order to adapt to changes will provide the regulator with a great decision-making power. The flexibility in the regulation will improve the correcting of the rules that were set in the beginning, being more efficient, which are based on acquired experience and achieved results. The structure of the agency and the flexibility of the regulation should be orientated on the promotion of competition in order to achieve economic and social development.

Um modelo de leilões com conteúdo local: análise e modelagem dos leilões de concessão de blocos exploratórios de petróleo e gás promovidos pela ANP no Brasil

Os leilões para concessão de blocos de petróleo no Brasil utilizam uma equação para formar a pontuação que define o vencedor. Cada participante deve submeter ao leiloeiro um lance composto por três atributos: Bônus de Assinatura (BA), Programa Exploratório Mínimo (PEM) e Conteúdo Local (CL). Cada atributo possui um peso na equação e a nota final de cada participante também depende dos lances ofertados pelos outros participantes. Apesar de leilões de petróleo serem muito estudados na economia, o leilão multi-atributos, do tipo máxima pontuação, ainda é pouco analisado, principalmente como mecanismo de alocação de direitos minerários. Este trabalho destaca a inserção do CL como atributo que transforma a estrutura, do que poderia ser um leilão simples de primeiro preço, em um leilão multi-atributos de máxima pontuação. Demonstra-se como o CL, através da curva de custos do projeto, está relacionado também ao Bônus de Assinatura, outro importante atributo da equação. Para compreender o impacto do fenômeno da inserção do CL, foram criados três casos de leilões hipotéticos, onde, dentre outras simplificações, o programa exploratório mínimo foi fixado para todas as empresas envolvidas. No caso base (Sem CL), simula-se a estrutura de um leilão de primeiro preço, onde apenas o BA define o vencedor do leilão. Já no caso forçado (CLO=CLR), há inserção do atributo CL, sendo o participante obrigado a cumprir o CL ofertado. Por fim, o caso completo (Com Multa) permite que o participante preveja a aplicação de multa por descumprimento do CL ofertado e, caso haja benefício econômico, descumpra efetivamente o CL ofertado. Considerando estes casos, argumenta-se que, apesar do o lucro das empresas e a eficiência do leilão não serem alterados, a inclusão do conteúdo local na estrutura do leilão pode ter reflexos consideráveis na receita do governo.

Os conflitos de competência normativa do estado regulador brasileiro no setor administrado pela ANP

The dissertation has by objective describe the administrative activity of regulation exercised by independent regulatory agencies, observing that this activity was already done before this structures creation, however, after a really deep administrative reform that had as objective built a Public Administration with more efficiency, it passed to be done with some own peculiarities of these new structures of regulation. The work gave especial attention to what concern the conflicts of normative competency that really often happen between ANP (Agência Nacional do Petróleo, Gás Natural e Biocombustíveis) and the legislatives organs of the Republic Federative of Brazil, because energetic area has unbelievable mater to any country, and the fact of some juridical norm be against the law and be accepted is very dangerous, it affronts the constitutional principle of the legality e may put in risk the democratic Estate of law, them, regulatory dogmatic must be scientifically knows, and developed, thought and especially there are so many doctrinaires divergences about regulation constitutionality. As a theorist point, the investigation got the Logical nocontradiction principle, according Hans Kelsen and Lourival Vilanova thought, doing a philosophical reflection about the system of positive law, in which there are many antinomies, or conflicts of norms, what include the conflicts of the administrative acts expedited by ANP and the legislation of the brazilian regulatory Estate. For a better understanding and exemplify some perplexities treated by the doctrinaire angle, this work did a lucubration about a possibility of a normative conflict between a ANP resolution and the municipal legislation in a specific case, also, brought several jurisprudences for the brazilians courts of justice, that confirm the empiric existence of normative conflicts among ANP s administrative norms and federal legislation. Finally, concludes observing that the regulation is not a legislative competency delegation to regulatory agencies, is just a new exercise of the administrative function, it is a technical specialization of the public administration, that using this know-how can acting with more efficiency, however the normative power of regulatory agencies must respect the empire of law, so in this terms, the dissertation suggests the ponderation of the constitutionals principles of efficiency and legality how form to harmonizing the democratic legitimate inherent to legal norm supremacy, with the perspective of an efficient economic and institutional development

Does plasma ANP participate in natriuresis induced by a-MSH?

-Melanocyte-stimulating hormone (-MSH; 0.6 and 3 nmol) microinjected into the anteroventral region of the third ventricle (AV3V) induced a significant increase in diuresis without modifying natriuresis or kaliuresis. Intraperitoneal (ip) injection of -MSH (3 and 9.6 nmol) induced a significant increase in urinary sodium, potassium and water excretion. Intraperitoneal (3 and 4.8 nmol) or iv (3 and 9.6 nmol) administration of -MSH did not induce any significant changes in plasma atrial natriuretic peptide (ANP), suggesting that the natriuresis, kaliuresis and diuresis induced by the systemic action of -MSH can be dissociated from the increase in plasma ANP. These preliminary results suggest that -MSH may be involved in a -MSHindependent mechanism of regulation of hydromineral metabolism.

Action of ANP on the nongenomic dose-dependent biphasic effect of aldosterone on NHE1 in proximal S3 segment

The rapid (2 min) nongenomic effects of aldosterone (ALDO) and/or spironolactone (MR antagonist), RU 486 (GR antagonist), atrial natriuretic peptide (ANP) and dimethyl-BAPTA (BAPTA) on the intracellular pH recovery rate (pHirr) via NHE1 (basolateral Na+/H+ exchanger isoform), after the acid load induced by NH4Cl, and on the cytosolic free calcium concentration ([Ca2+](i)) were investigated in the proximal S3 segment isolated from rats, by the probes BCECF-AM and FLUO-4-AM, respectively. The basal pHi was 7.15+/-0.008 and the basal pHirr was 0.195+/-0.012 pH units/min (number of tubules/number of tubular areas = 16/96). Our results confirmed the rapid biphasic effect of ALDO on NHE1: ALDO (10(-12) M) increases the pHirr to approximately 59% of control value, and ALDO (10(-6)M) decreases it to approximately 49%. Spironolactone did not change these effects, but RU 486 inhibited the stimulatory effect and maintained the inhibitory effect. ANP (10(-6) M) or BAPTA (5 x 10(-5) M) alone had no significant effect on NHE1 but prevented both effects of ALDO on this exchanger. The basal [Ca2+](i) was 104+/-3 nM (15), and ALDO (10(-12) or 10(-6) M) increased the basal [Ca2+](i) to approximately 50% or 124%, respectively. RU 486, ANP and BAPTA decreased the [Ca2+](i) and inhibited the stimulatory effect of both doses of ALDO. The results suggest the involvement of GR on the nongenomic effects of ALDO and indicate a pHirr-regulating role for [Ca2+](i) that is mediated by NHE1, stimulated/impaired by ALDO, and affected by ANP or BAPTA with ALDO. The observed nongenomic hormonal interaction in the S3 segment may represent a rapid and physiologically relevant regulatory mechanism in the intact animal under conditions of volume alterations. (C) 2011 Elsevier Ltd. All rights reserved.

ANP impairs the dose-dependent stimulatory effect of ANG II or AVP on H+-ATPase subcellular vesicle trafficking

The effect of angiotensin II (ANG II) or arginine vasopressin (AVP) alone or plus atrial natriuretic peptide (ANP) on H+-ATPase subcellular vesicle trafficking was investigated in MDCK cells following intracellular pH (pHi) acidification by exposure to20 mMNH4Cl for 2 min in a Na+-free solution containing Schering 28080, conditions under which H+-AT-Pase is the only cell mechanism for pHi recovery. Using the acridine orange fluorescent probe (5mM) and confocal microscopy, the vesicle movement was quantified by determining, for each experimental group, the mean slope of the line indicating the changes in apical/basolateral fluorescence density ratio over time during the first 5.30 min of the pHi recovery period. Under the control conditions, the mean slope was 0.079 ± 0.0033 min-1 (14) and it increased significantly with ANG II [10-12 and 10-7 M, respectively to 0.322 ± 0.038 min-1 (13) and 0.578 ± 0.061 min-1 (12)] or AVP [10-12 and 10-6 M, respectively to 0.301 ± 0.018 min-1 (12) and 0.687 ± 0.049 min-1 (11)]. However, in presence of ANP (10-6 M, decreases cytosolic free calcium), dimethyl-BAPTA/AM (5 × 10-5 M, chelates intracellular calcium) or colchicine (10-5 M, 2-h preincubation; inhibits microtubule-dependent vesicular trafficking) alone or plus ANG II or AVP the mean slopes were similar to the control values, indicating that such agents blocked the stimulatory effect of ANG II or AVP on vesicle trafficking. The results suggest that the pathway responsible for the increase in cytosolic free calcium and the microtu-bule-dependent vesicular trafficking are involved in this hormonal stimulating effect. Whether cytosolic free calcium reduction represents an important direct mechanism for ANP impairs the dose-dependent stimulatory effect of ANG II or AVP on H+-ATPase subcellular vesicle trafficking, or is a side effect of other signaling pathways which will require additional studies.