Abnormal regulation of Na,K-ATPase in Glucose Intolerant Rats.

Introduction: Glucose is the most important physiological insulin secretagogue. However, the mechanisms underlying glucose-induced insulin release are not fully understood. The role of electrogenic systems such as ionic pumps, to these events remains essentially uninvestigated. Na,K-ATPase, responsible for maintaining Na+ and K+ gradients across the plasma membrane and generates a net outward current, thus changes in its activity may contribute to the early ionic events regulating insulin secretion (Therien and Blostein, 2000). Objective: The aim of this work was to evaluate the regulation of Na,K-ATPase activity by glucose in intact -cells of normal and glucose intolerant (GI) rats and its putative contribution to the regulation of insulin secretion. Material and Methods: Pancreatic -cells, from normal or control or GI rats, were isolated and cultured (48h). Cell batches were pre-incubated (30min) with 2mM glucose to reach basal. Afterwards cells were challenged with glucose in the interval 0-11mM for 60min, for dose-dependence evaluation, or with 8mM glucose for 5-120min, for time-dependence evaluation. ATPase activity was assessed in intact cells by colorimetric quantification of Pi formed in 30min. Na,K-ATPase activity was calculated by the difference between the activities obtained in the absence and in presence the of 1mM ouabain (Costa et al., 2009). Results: In β-cells from normal rats, glucose induced a bimodal regulation of Na,K-ATPase. In the absence of glucose, Na,K-ATPase activity was 0.056±0.015 U/mg. Stimulation with 2mM glucose induced an increase of Na,K-ATPase activity of ~4 fold whereas for [glucose] above 2mM it was observed a significant inhibition of Na,K-ATPase activity (0.061±0.013, 0.080±0.009 and 0.064±0.005 U/mg for 5.6, 8.4 and 11mM glucose, respectively, compared to 0.188±0.035 U/mg observed in 2mM G; n=3-8). β-cells from GI rats does not present this profile; in the absence of glucose, Na,K-ATPase activity was 0.202±0.036 U/mg and no significant differences from this value were observed with the other glucose concentration tested. Addicionally, in β-cells from normal rats, glucose (8mM) induced a time-dependent inhibition, with a biphasic profile, of Na,K-ATPase - it was observed a decrease in the pump activity between 0 and 20min stimulation where it reached a minimum value (77%). For incubation periods over 20min, the pump activity slowly and partially recovered (54%, 55% and 52%, for 30, 60 and 120min, respectively; n=7). In β-cells from GI animals, an less accentuated decrease of Na,K-ATPase activity between 0 ans 20min was also observed (34%), and is not observed further recover in activity. Conclusions: This work demonstrates there Na,K-ATPase is strictly regulated by glucose in pancreatic β-cell. This regulation is unpaired in GI animals. Na,K-ATPase contribution to glucose-induced ionic events and insulin secretion might be relevant and must be explored as a possible therapeutic target in TD2 . 1. Therien AG, Blostein R (2000) Mechanisms of sodium pump regulation. Am J Physiol Cell Physiol 279:C541-C566 2. Costa AR, Real J, Antunes CM, Cruz-Morais J (2009) A new approach for determination of Na,K-ATPase activity: application to intact pancreatic beta-cells. In Vitro Cell Dev Biol Ani

Implication of AMPK in glucose-evoked modulation of Na,K-ATPase

Background and aims: Na,K-ATPase is an integral membrane protein that maintains the gradients of Na+ and K+, using the energy of ATP hydrolysis, maintaining the ionic gradients that allow electrical activity to occur. It has been demonstrated that, in pancreatic β-cells, Na,K-ATPase is regulated by glucose and that this phenomenon is impaired in glucose intolerant subjects. However, the mechanism underlying glucose-induced modulation of Na,K-ATPase is still unclear. The AMP-activated protein kinase (AMPK) is a molecular key player in energy homeostasis, providing exquisite sensitivity to small changes in intracellular AMP levels and thus to intracellular [ATP]/[ADP] ratio, that is known to activate protein regulatory pathways. Since in pancreatic β-cell, glucose has marked effects on oxidative metabolism and total intracellular ATP and AMP levels, the involvement of AMPK in the cascade of events regulating Na,K-ATPase regulation in pancreatic β-cells was postulated. The aim of this work was to evaluate the putative role of AMPK in the glucose-evoked regulation of Na,K-ATPase activity in the pancreatic β-cell. Materials and methods: Pancreatic -cells from normal (control) or glucose-intolerant Wistar rats (GIR) were isolated and cultured (48h). Cell batches were pre-incubated (30min) with 2.1mM glucose to reach basal activity. Afterwards cells were challenged to 8.4mM glucose for 20min, in the presence or absence of AMPK agonists (AICAR) and antagonists (compound C; CC). ATPase activity was assessed in intact cells by colorimetric quantification of Pi formed in 30min. Na,K-ATPase activity was calculated by the difference between the activities obtained in the absence and in presence the of 1mM ouabain. Results: In basal conditions the activity of Na,K-ATPase from normal and GIR pancreatic β-cell was similar (0.184±0.030 and 0.186±0.020 molPi/min/mgProt, respectively). Challenging the control β-cells with glucose 8.4mM evoked a 62% reduction of Na,K-ATPase activity whereas in GIR β-cells a significantly lower inhibition (40%) was observed. The addition of AICAR 1mM abolished glucose-induced Na,K-ATPase inhibition (0,166±0.011 molPi/min/mg). In control β-cell, the addition of CC 10 μM had no effect on glucose-induced inhibition of Na,K-ATPase. In the contrary, in GIR β-cells it significantly potentiated glucose-evoked inhibition of Na,K-ATPase reaching values similar to that observed in the controls (66%). Conclusions: The AMPK agonist AICAR counteracts the inhibitory action of glucose on Na,K-ATPase of control β-cells whereas CC amplified the glucose-induced inhibition of Na,K-ATPase in GIR β-cells. These results suggest that AMPK plays a central role in the cascade of events underlying glucose-induced modulation of Na,K-ATPase and that the defect must be upstream of AMPK. Finally, abnormal glucose-induced regulation of Na,K-ATPase occurs prior to overt type 2 diabetes and might be a feature in the disease development

Chemical composition and anti-diabetic properties of Cytisus multiflorus

Bakground and aims: The interest on plants with potential medicinal properties has been increasing worldwide. In the Iberian Peninsula there are some endemic species known by the population for their pharmacologic activity with valorization potential that have not been yet characterized. The white Spanish broom (Cytisus multiflorus) is described as having anti-diabetic effect [1] and in a preliminary the hypoglycemic and hyper-insulinemic effect of an aqueous extract has been shown [2]. The aim of this work was to fractionate and analyse the composition of the aqueous extract of C. multiflorus flowering parts and evaluate its potential as an anti-diabetic agent. Materials and methods: The aqueous extract was primarily fractionated by SPE using water:methanol (W:Me) eluent (a 10% step-wise gradient W:Me from 100:0 to 0:100) followed by high performance liquid chromatography with diode array detector (HPLC-DAD). The most relevant fraction were analysed by LC-MS to determine the chemical composition. Total fenol content was determined by a modified Folin-Ciocalteau method and the anti-oxidant activity was evaluated by the DPPH mehod. Finally, the hipoglicemic potential was evaluated in vivo using glucose intolerant rats (GIR). Results: Eleven fractions of the bulk extract were obtained. Seven of these fractions (10, 30, 40, 50, 60, 70 e 80% Me) were found to have a relevant compounds, mostly flavonoid compounds, namely, rutin (50, 60 and 70% Me fractions), ferrulic acid (30% Me), referred as having hypoglicemic effect. The fractions obtained with 50 and 70% Me showed the highest content in phenol equivalents and the highest anti-oxidant effect were found in the 50 and 60% Me fractions. The 30 and 60% Me fraction had no effect on the post-prandial glicemia. Conclusions: The 30, 50, 60 and 70% Me fractions, due to their chemical composition and anti-oxidant effects were the most promising to have anti-diabetic effect. However, the 30 and 60% Me were found to be ineffective. The 50% Me fraction showed both a high content of flavonoid compounds and the highest anti-oxidant power which suggest that it may constitute the most promising one. The anti-diabetic properties of this fraction should be investigated. [1] Camejo-Rodrigues J. et al. (2003). J. Ethnopharmacol, 89, 199-209 [2] Célia M. Antunes, Laurinda R. Areias, Inês P. Vieira, Ana C. Costa, M. Teresa Tinoco, & Júlio Cruz-Morais (2009). Rev. Fitoterapia 9 (Supl.1): 91

Alterações na modulação da Na,K-ATPase pela glicose em célula β-pancreática de animais intolerantes à glicose.

Alterações na modulação da Na,K-ATPase pela glicose em célula β-pancreática de animais intolerantes à glicose AR Costa1,2, CM Antunes1,3, J Cruz-Morais1,2 1Departamento de Química, 2Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), Universidade de Évora, Largo dos Colegiais 2, 7000 Évora; 3Centro de Neurociências e Biologia Celular (CNC), Universidade de Coimbra, 3004-517 Coimbra;[email protected] A diabetes tipo 2 é uma síndrome metabólica, que se caracteriza por alterações da secreção ou acção da insulina, cujas bases bioquímicas não estão completamente esclarecidas. Na célula-, a resposta fisiológica à glicose envolve a despolarização, resultante principalmente do fecho de canais de K+ sensíveis ao ATP, seguido do influxo de Ca2+ por canais sensíveis à voltagem tipo-L e consequente aumento da concentração intracelular de Ca2+, necessário à exocitose da insulina, hormona responsável pela homeostasia da glicose. A Na,K-ATPase (ou bomba de sódio), é o sistema de transporte activo responsável por manter o gradiente transmembranar de Na+ e K+. Estudos recentes têm sugerindo que a inibição da Na,K-ATPase possa contribuir para a despolarização induzida por glicose. Como o acoplamento estímulo-secreção se encontra alterado em célula- de animais diabéticos, proposemo-nos estudar a participação da Na,K-ATPase neste mecanismo utilizando um modelo de pré-diabetes que se caracteriza essencialmente por apresentar alterações na curva de tolerância à glicose. Para isso, desenvolveu-se um método que permite avaliar a actividade da bomba em células β-pancreáticas primárias, mantidas em culturas aderentes, que preservam os processos de transdução de sinal. Com este método observámos que a glicose inibe a Na,K-ATPase em animais normais, estando este processo significativamente atenuado em animais pré-diabéticos. Observámos ainda que a secreção de insulina estimulada por glicose, nas células de pré-diabéticos, é cerca de 4 vezes menor relativamente aos controlos. Estes dados permitem-nos sugerir que a regulação da bomba por glicose se encontra modificada em animais pré-diabéticos, podendo contribuir para as alterações dos mecanismos iónicos responsáveis pelo acoplamento estímulo-secreção, nomeadamente na cinética da despolarização, comprometendo a secreção de insulina. O trabalho prossegue no sentido de esclarecer os mecanismos através dos quais a glicose induz inibição na Na,K-ATPase, bem como no levantamento dos factores que podem contribuir para as diferenças observadas entre os animais normais e pré-diabéticos, nomeadamente ao nível da expressão isoenzimática

A review of data mining applications in semiconductor manufacturing

The authors acknowledge Fundacao para a Ciencia e a Tecnologia (FCT-MCTES) for its financial support via the project UIDB/00667/2020 (UNIDEMI).For decades, industrial companies have been collecting and storing high amounts of data with the aim of better controlling and managing their processes. However, this vast amount of information and hidden knowledge implicit in all of this data could be utilized more efficiently. With the help of data mining techniques unknown relationships can be systematically discovered. The production of semiconductors is a highly complex process, which entails several subprocesses that employ a diverse array of equipment. The size of the semiconductors signifies a high number of units can be produced, which require huge amounts of data in order to be able to control and improve the semiconductor manufacturing process. Therefore, in this paper a structured review is made through a sample of 137 papers of the published articles in the scientific community regarding data mining applications in semiconductor manufacturing. A detailed bibliometric analysis is also made. All data mining applications are classified in function of the application area. The results are then analyzed and conclusions are drawn.publishersversionpublishe

Economic burden of personal protective strategies for dengue disease: an optimal control approach

Dengue fever is a vector-borne disease that is widely spread. It has a vast impact on the economy of countries, especially where the disease is endemic. The associated costs with the disease comprise prevention and treatment. This study focus on the impact of adopting individual behaviors to reduce mosquito bites - avoiding the disease’s transmission - and their associated costs. An epidemiological model is presented with human and mosquito compartments, modeling the interaction of dengue disease. The model assumed some self-protection measures, namely the use of repellent in human skin, wear treated clothes with repellent, and sleep with treated bed nets. The household costs for these protections are taking into account to study their use. We conclude that personal protection could have an impact on the reduction of the infected individuals and the outbreak duration. The costs associated with the personal protection could represent a burden to the household budget, and its purchase could influence the shape of the infected’s curve.publishe

Substantive rationality of impression management:an analysis of corporate reports of JBS

Objective: To investigate the substantive rationality inherent to the practice of impression management (IM). Originality/value:Based on the taxonomy created by Merkl-Davies and Brennan (2011), it is analyzed: 1. the discussion under the sociological perspective; and 2. the actions of substantive rationality in the management of impressions that can manifest through actions of symbolic management. This study responds to the call of Merkl-Davies et al. (2011) by putting into practice an unconventional analytical model based on the use of the corporate reports (annual and sustainability) of the company JBS with special attention to the “Carne Fraca” [Weak Meat] Operation. Design/methodology/approach: A content analysis of the accounting narratives of JBS’ corporate reports between 2015 and 2018 was carried out.Findings: Accounting narratives suggest evidence of IM and substantive rationality. The preparers of the corporate reports used substantive rationality to legitimize the company’s actions, with the most significant manifestations of this practice in the 2017 report, the year of the “Carne Fraca” operation. The implications of this research lie in demonstrating evidence that guides companies regarding the use of accounting narratives to convince society of their actions as consistent with the structure of social rules and norms. Furthermore, we emphasize that the accounting narratives must be analyzed with caution since they can be elaborated under the influence of the manager’s opportunistic behavior.info:eu-repo/semantics/publishedVersio

Impact of frost on the morphology and chemical composition of cv. Santulhana olives

Frost events and extreme weather phenomena greatly affect several characteristics of the olive fruit. This study evaluated the impact of frost on the morphology, composition (moisture, fat, fatty acids, tocopherols, and total phenolic contents), and antioxidant activity of olives of cv. Santulhana. A total of 14 trees from the same geographical region (Santulhão, northeast of Portugal) were chosen, including trees subjected or not subjected to frost conditions (n = 7 each). The results showed that frost led to morphological changes in olive fruits, particularly in terms of weight and diameter, which were imposed by a huge decrease in the moisture content (-20%). Fat relative content increased as a consequence of the water loss (+29% in fresh pulp weight), with a slight reduction of the relative abundance of saturated fatty acids (-4%) and tocopherol contents in the fat (-17%). However, the total phenolic contents and antioxidant activity were severely affected (-70% and -42%, respectively), with potential consequences for the olive oil stability and sensorial attributes. Principal component analysis showed that both morphological and chemical parameters could be used as biomarkers to identify olives subjected or not subjected to frost. The overall negative impact of frost on the minor antioxidant contents of cv. Santulhana olives may anticipate a quality loss of olive oils extracted from olives affected by frost.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020) and to REQUIMTELAQV (UIDB/50006/2020) units and to the Associate Laboratories for Green Chemistry-LAQV (UIDB/50006/2020) and SusTEC (LA/P/0007/2020). Nuno Rodrigues thanks to National funding by FCT—Foundation for Science and Technology, P.I., through the institutional scientific employment program-contract. Ana I. Rodrigues thanks the financial support by OleaChain project “Skills for sustainability and innovation in the value chain of traditional olive groves in the Northern Interior of Portugal” (NORTE-06-3559-FSE-000188).info:eu-repo/semantics/publishedVersio