Self-association of isolated large cytoplasmic domain of plasma membrane H+-ATPase from Saccharomyces cerevisiae: Role of the phosphorylation domain in a general dimeric model for P-ATPases

AbstractLarge cytoplasmic domain (LCD) plasma membrane H+-ATPase from S. cerevisiae was expressed as two fusion polypeptides in E. coli: a DNA sequence coding for Leu353–Ileu674 (LCDh), comprising both nucleotide (N) and phosphorylation (P) domains, and a DNA sequence coding for Leu353–Thr543 (LCDΔh, lacking the C-terminus of P domain), were inserted in expression vectors pDEST-17, yielding the respective recombinant plasmids. Overexpressed fusion polypeptides were solubilized with 6 M urea and purified on affinity columns, and urea was removed by dialysis. Their predicted secondary structure contents were confirmed by CD spectra. In addition, both recombinant polypeptides exhibited high-affinity 2′,3′-O-(2,4,6-trinitrophenyl)adenosine-5′-triphosphate (TNP-ATP) binding (Kd=1.9 μM and 2.9 μM for LCDh and LCDΔh, respectively), suggesting that they have native-like folding. The gel filtration profile (HPLC) of purified LCDh showed two main peaks, with molecular weights of 95 kDa and 39 kDa, compatible with dimeric and monomeric forms, respectively. However, a single elution peak was observed for purified LCDΔh, with an estimated molecular weight of 29 kDa, as expected for a monomer. Together, these data suggest that LCDh exist in monomer–dimer equilibrium, and that the C-terminus of P domain is necessary for self-association. We propose that such association is due to interaction between vicinal P domains, which may be of functional relevance for H+-ATPase in native membranes. We discuss a general dimeric model for P-ATPases with interacting P domains, based on published crystallography and cryo-electron microscopy evidence

The slope current along the western European margin: A numerical investigation

A three-dimensional hydrodynamic model is used to investigate the poleward flow along the western European slopes. The area of the model domain goes from northwest Africa to Ireland. During a first stage, the currents are driven by climatological density fields. In a second stage the model is also forced by climatological winds, although special attention is paid to spring, when the winds are favourable to the development of an equatorward jet along the Iberian coast. Results show that the climatological density field is able to produce the poleward current along the European continental slope. Winds can modify the flow pattern, mainly in the southern areas off Iberia, specially during the upwelling season. The poleward current obtained is continuous between the Portuguese and French coasts, following the contour depths, with a core of maximum velocity located from 300-1 500 m, depending both on space and time. This core corresponds to the shelfward divergence of isopycnals. Maximum speeds range from 10-20 cm/s. A poleward intensification of the current is also obtained.Un modelo tridimensional hidrodinámico se ha usado para investigar el flujo en dirección hacia el polo a lo largo del talud occidental europeo. El área de dominio del modelo va desde el noroeste de áfrica hasta Irlanda. En un primer estadio, las corrientes son dirigidas por los campos climatológicos de densidad. En un segundo estadio el modelo se fuerza con vientos climatológicos, donde se presta una especial atención a la primavera, cuando los vientos son favorables al desarrollo de un jet en dirección al ecuador a lo largo de la costa ibérica. Los resultados muestran cómo el campo de densidad climatológica es capaz de producir una corriente en dirección al polo a lo largo del talud del continente europeo. Los vientos pueden modificar este esquema de flujo, principalmente en las áreas al sur de la península Ibérica, sobre todo durante la estación de upwelling. La corriente hacia el polo obtenida es continua entre las costas portuguesa y francesa, siguiendo las líneas de profundidad, con una velocidad máxima en el núcleo situada entre 300 m y 1 500 m, dependiendo tanto del espacio como del tiempo. Este núcleo corresponde a la divergencia de las isopicnas hacia la plataforma continental. Las velocidades máximas varían entre 10 y 20 cm/s. También se obtiene una intensificación hacia el polo de la corriente.Instituto Español de Oceanografí

CardNutri:A Software of Weekly Menus Nutritional Elaboration for Scholar Feeding Applying Evolutionary Computation

This paper aims to present and evaluate a software that uses an evolutionary strategy to design weekly nutritional menus for School Feeding. The software ensures the nutritional needs of students and also minimizes the total cost of the menu. We based our nutritional needs on the Brazilian National School Feeding Programme (PNAE). This program takes into account: (i) the age of the student; (ii) some preparations issues as color, consistency and, variety; and also (iii) the maximum amount to be paid per meal. Our software generates, in less than five minutes, a set of menus, and the nutritionist can choose the menu that suits his/her best. We evaluate our algorithm using the Weighted-Sum approach, and our results show that the obtained 5-days menus using the proposed methodology not only comply with the restrictions imposed by the authorities but also produce inexpensive and healthy menus. We also appraise the software itself using an opinion pool among nine nutritionists. The professionals considered our software above expectations

Pro-oxidant Activity And Genotoxicity Of The Astronium Fraxinifolium Using Wing Smart And Allium Cepa Test

Astronium fraxinifolium is an arboreal species found throughout the Brazilian Cerrado region and used in folk medicine as antimicrobial, anti-hemorrhagic and healing. Pro-oxidant activity of extracts of fraxinifolium rough Relative Electrophoresis Mobility (REM) of Bovine Serum Albumin (BSA) protein in presence of the extract and Cu2+ and also the genotoxic potential through Somatic Mutation and Recombination Test (SMART) and Allium cepa tests has been investigated during this study. In the REM, the extracts acting exclusively on BSA resulted in a band formation with higher molecular weight than BSA, probably due to oxidative action of the extracts. In presence of Cu2+and extracts occurred protein fragmentation due to Cu2+ oxidative action potentiated by the extracts. In SMART test the frequency of mutant spots increases with the increased concentration of extract A. fraxinifolium (50 mg mL-1= 1.40 and 100 mg mL-1 = 2.66). The HB cross shows a decrease in the total of mutant spots frequency for the different treatments (50 mg mL-1 =6.60 and 100 mg mL-1 = 3.25). Evaluation of A cutest demonstrated the following results (concentration extract = chromosomal abnormalities) 1 mg mL-1 = 85,10 mg L-1 = 61 50 mg mL-1 = 53,100 mg mL-1 =33 and MMS 10 mg L-1 = 50. Genotoxic and cytotoxic actions can be explained by the actions of tannins present in its composition. Butthere may be other substances that also act for such results. The genotoxicity in medicinal plants contributes to therapeutic safety warning that although the use of medicinal plants is an inexpensive and non-aggressive method it can cause harmful effects if used incorrectly. © 2016 Rafael Rozolen Teixeria Zafred et al.10427628

Effect Of Post Type And Restorative Techniques On The Strain And Fracture Resistance Of Flared Incisor Roots

Restoring flared endodontically treated teeth continues to be a challenge for clinicians. This study evaluated the effect of post types and restorative techniques on the strain, fracture resistance, and fracture mode of incisors with weakened roots. One hundred five endodontically treated bovine incisors roots (15 mm) were divided into 7 groups (n=15). The two control groups were (C) intact roots restored with Cpc (cast posts and core) or Gfp (glass fiber posts). The five experimental groups were (F) flared roots restored with GfpAp (Gfp associated with accessory glass fiber posts), GfpRc (anatomic Gfp, relined with composite resin), and GfpRcAp (anatomized Gfp with resin and accessory glass fiber posts). All teeth were restored with metal crowns. Mechanical fatigue was performed with 3x10 5/50 N. Specimens were loaded at 45°, and the strain values (μS) were obtained on root buccal and proximal surfaces. Following that, the fracture resistance (N) was measured. One-way ANOVA and Tukey's HSD tests (α=0.05) were applied, and failure mode was checked. No significant difference in strain values among the groups was found. Cpc presented lower fracture resistance and more catastrophic failures in flared roots. Gfp associated with composite resin or accessory glass fiber posts seems to be an effective method to improve the biomechanical behavior of flared roots.223230237Coelho, C.S., Biffi, J.C., Silva, G.R., Abrahão, A., Campos, R.E., Soares, C.J., Finite element analysis of weakened roots restored with composite resin and posts (2009) Dent Mater J, 28, pp. 671-678Teixeira, C.S., Silva-Sousa, Y.T., Sousa-Neto, M.D., Bond strength of fiber posts to weakened roots after resin restoration with different light-curing times (2009) J Endod, 35, pp. 1034-1039Zogheib, L.V., Pereira, J.R., do Valle, A.L., de Oliveira, J.A., Pegoraro, L.F., Fracture resistance of weakened roots restored with composite resin and glass fiber post (2008) Braz Dent J, 19, pp. 329-333Marchi, G.M., Mitsui, F.H., Cavalcanti, A.N., Effect of remaining dentine structure and thermal-mechanical aging on the fracture resistance of bovine roots with different post and core systems (2008) Int Endod J, 41, pp. 969-976da Silveira, T.C., Santos, F.M.C., Silva-Sousa, Y.T., de Sousa-Neto, M.D., Interfacial evaluation of experimentally weakened roots restored with adhesive materials and fibre posts: An SEM analysis (2008) J Dent, 36, pp. 672-682Moosavi, H., Maleknejad, F., Kimyai, S., Fracture resistance of endodontically-treated teeth restored using three root- reinforcement methods (2008) J Contemp Dent Pract, 9, pp. 30-37Bonfante, G., Kaizer, O.B., Pegoraro, L.F., do Valle, A.L., Fracture strength of teeth with flared root canals restored with glass fibre posts (2007) Int Dent J, 57, pp. 153-160Santos, A.F., Meira, J.B., Tanaka, C.B., Xavier, T.A., Ballester, R.Y., Lima, R.G., Can fiber posts increase root stresses and reduce fracture? (2010) J Dent Res, 89, pp. 587-591da Silva, N.R., Raposo, L.H., Versluis, A., Fernandes-Neto, A.J., Soares, C.J., The effect of post, core, crown type, and ferrule presence on the biomechanical behavior of endodontically treated bovine anterior teeth (2010) J Prosthet Dent, 104, pp. 306-317Silva, N.R., Castro, C.G., Santos-Filho, P.C., Silva, G.R., Campos, R.E., Soares, P.V., Influence of different post design and composition on stress distribution in maxillary central incisor: Finite element analysis (2009) Indian J Dent Res, 20, pp. 153-158Santos-Filho, P.C., Castro, C.G., Silva, G.R., Campos, R.E., Soares, C.J., Effects of post system and length on the strain and fracture resistance of root filled bovine teeth (2008) Int Endod J, 41, pp. 493-501Soares, C.J., Castro, C.G., Santos, F.P.C., Soares, P.V., Magalhaes, D., Martins, L.R., Two-dimensional FEA of dowels of different compositions and external surface configurations (2009) J Prosthodont, 18, pp. 36-42Macedo, V.C., de Faria, S.A.L., Martins, L.R., Effect of cement type, relining procedure, and length of cementation on pull-out bond strength of fiber posts (2010) J Endod, 36, pp. 1543-1546Leitune, V.C., Collares, F.M., Werner, S.S.M., Influence of chlorhexidine application at longitudinal push-out bond strength of fiber posts (2010) Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 110, pp. 77-81Novais, V.R., Quagliatto, P.S., Bona, A.D., Correr-Sobrinho, L., Soares, C.J., Flexural modulus, flexural strength, and stiffness of fiber-reinforced posts (2009) Indian J Dent Res, 20, pp. 277-281Marchi, G.M., Paulillo, L.A., Pimenta, L.A., de Lima, F.A., Effect of different filling materials in combination with intraradicular posts on the resistance to fracture of weakened roots (2003) J Oral Rehabil, 30, pp. 623-629Soares, C.J., Pizi, E.C., Fonseca, R.B., Martins, L.R., Influence of root embedment material and periodontal ligament simulation on fracture resistance tests (2005) Braz Oral Res, 19, pp. 11-16Huysmans, M.C., Peters, M.C., van der Varst, P.G., Plasschaert, A.J., Failure behaviour of fatigue-tested post and cores (1993) Int Endod J, 26, pp. 294-300Soares, P.V., Santos-Filho, P.C., Gomide, H.A., Araujo, C.A., Martins, L.R., Soares, C.J., Influence of restorative technique on the biomechanical behavior of endodontically treated maxillary premolars. Part II: Strain measurement and stress distribution (2008) J Prosthet Dent, 99, pp. 114-122Kishen, A., Kumar, G.V., Chen, N.N., Stress-strain response in human dentine: Rethinking fracture predilection in post-core restored teeth (2004) Dent Traumatol, 20, pp. 90-10

Identification of proteomic signatures associated with depression and psychotic depression in post-mortem brains from major depression patients

Major depressive disorder (MDD) is a leading cause of disability worldwide and results tragically in the loss of almost one million lives in Western societies every year. This is due to poor understanding of the disease pathophysiology and lack of empirical medical tests for accurate diagnosis or for guiding antidepressant treatment strategies. Here, we have used shotgun proteomics in the analysis of post-mortem dorsolateral prefrontal cortex brain tissue from 24 MDD patients and 12 matched controls. Brain proteomes were pre-fractionated by gel electrophoresis and further analyzed by shotgun data-independent label-free liquid chromatography-mass spectrometry. This led to identification of distinct proteome fingerprints between MDD and control subjects. Some of these differences were validated by Western blot or selected reaction monitoring mass spectrometry. This included proteins associated with energy metabolism and synaptic function and we also found changes in the histidine triad nucleotide-binding protein 1 (HINT1), which has been implicated recently in regulation of mood and behavior. We also found differential proteome profiles in MDD with (n=11) and without (n=12) psychosis. Interestingly, the psychosis fingerprint showed a marked overlap to changes seen in the brain proteome of schizophrenia patients. These findings suggest that it may be possible to contribute to the disease understanding by distinguishing different subtypes of MDD based on distinct brain proteomic profiles

Variance fluctuations in nonstationary time series: a comparative study of music genres

An important problem in physics concerns the analysis of audio time series generated by transduced acoustic phenomena. Here, we develop a new method to quantify the scaling properties of the local variance of nonstationary time series. We apply this technique to analyze audio signals obtained from selected genres of music. We find quantitative differences in the correlation properties of high art music, popular music, and dance music. We discuss the relevance of these objective findings in relation to the subjective experience of music.Comment: 13 pages, 4 fig

Proteomic changes in serum of first onset, antidepressant drug-naïve major depression patients

Major depressive disorder (MDD) is a complex and multi-factorial disorder. Although genetic factors and other molecular aspects of MDD have been widely studied, the underlying pathological mechanisms are still mostly unknown. We sought to investigate the pathophysiology of MDD by identifying and characterising serum molecular differences and their correlation to symptom severity in first onset, antidepressant drug-naïve MDD patients. We performed an exploratory molecular profiling study on serum samples of MDD patients and controls using multiplex immunoassay and label-free liquid chromatography mass spectrometry in data independent mode (LC-MSE). We included two independent cohorts of first onset, antidepressant drug-naïve MDD patients (n = 23 and 15) and matched controls (n = 42 and 21) in our study in order to validate the results. The main outcome included the following list of circulatory molecules changing and/or correlating to symptom severity: angiotensin-converting enzyme, acute phase proteins (e.g. ferritin and serotransferrin), brain-derived neurotrophic factor, complement component C4-B, cortisol, cytokines (e.g. macrophage migration inhibitory factor and interleukin-16), extracellular newly identified receptor for advanced glycosylation end products-binding protein, growth hormone and superoxide dismutase-1. This study provides evidence of an increased pro-inflammatory and oxidative stress response, followed by a hyperactivation of the HPA-axis in the acute stages of first onset MDD, as well as a dysregulation in growth factor pathways. These findings help to elucidate MDD related pathways in more detail and further studies may lead to identification of novel drug targets, inc

Effect on Broiler Production Performance and Meat Quality of Feeding Ulva lactuca Supplemented with Carbohydrases

Macroalgae have been increasingly exploited worldwide for feed, food and biofuel applications, due to their nutritive and bioactive compounds. Green seaweeds belonging to the genus Ulva have high growth rates, which makes them suitable for being cultured in sustainable algae production, such as an integrated multi-trophic aquaculture system. This is expected to increase the use of Ulva sp. as an alternative source to conventional feedstuffs (e.g., cereals and soybean meal) for poultry. The objective of the current study was to assess if the incorporation of 15% Ulva lactuca in broiler chickens’ diet, combined or not with carbohydrate-active enzymes, would enhance meat nutritional quality without compromising animal growth performance. Overall, U. lactuca led to an accumulation of antioxidant carotenoids, n-3 PUFA and macrominerals, including magnesium, potassium and phosphorus, in the breast muscle, with likely health benefits, without significantly impairing growth performance. The supplementation of macroalgae with a recombinant ulvan lyase reduced ileal viscosity with possible beneficial effects on broiler digestibility. Although dietary U. lactuca showed potential to increase meat quality, it reduced meat overall acceptability, which suggests the use of a lower algae inclusion level to prevent a negative meat sensory perception for consumersinfo:eu-repo/semantics/publishedVersio

Estudo do complexo valvar atrioventricular cardíaco esquerdo em búfalos (Bubalus bubalis) da raça Jafarabadi

Atrioventricular valve complex of 30 Jafarabadi water buffaloes, adult males were studied in this research with no heart diseases. The animals were obtained from a slaughterhouse in Brazilian State of Parana. The hearts were opened at the third portion affording access to the valve complex. The complexes had its area, number and type of tendinous cords submitted to analysis. The results showed that the complex is composed by two cusps and four accessory cusps, two or three papillary muscles in which 10-25 tendinous cords fix on the cusps that face the ventricle wall. The total area of the complex was on average 38.56cm², with a minimum of 24.96cm² and a maximum of 55.54cm². Statistically, no relation between the number of cords and the cusps' area where they are inserted or with the number of papillary muscle where they originated from was observed.Foram estudados os complexos valvares atrioventricular esquerdo de 30 búfalos da raça Jafarabadi, machos e adultos, sem alterações cardíacas, provenientes de abatedouros do Estado do Paraná. Os corações foram examinados em seu terço médio para acesso ao complexo valvar, que foi submetido a estudos de área, número e tipificação de cordas tendíneas. Os resultados demonstram que este complexo é formado por duas cúspides principais e quatro cúspides acessórias, apresentam em sua formação de 2-3 músculos papilares, nos quais se inserem de 10-25 cordas tendíneas, que se fixam em cúspides voltadas para a parede do ventrículo. A área total deste complexo apresenta uma média de 38,56cm² com um mínimo de 24,96cm² e um máximo de 55,54cm². Estatisticamente não há relação entre número de cordas e a área da cúspide onde estas estão inseridas, nem com o número de músculos papilares dos quais elas provem