Bothrops insularis venomics: A proteomic analysis supported by transcriptomic-generated sequence data

A joint transcriptomic and proteomic approach employing two-dimensional electrophoresis, liquid chromatography and mass spectrometry was carried out to identify peptides and proteins expressed by the venom gland of the snake Bothrops insularis, an endemic species of Queimada Grande Island, Brazil. Four protein families were mainly represented in processed spots, namely metalloproteinase, serine proteinase, phospholipase A(2) and lectin. Other represented families were growth factors, the developmental protein G10, a disintegrin and putative novel bradykinin-potentiating peptides. The enzymes were present in several isoforms. Most of the experimental data agreed with predicted values for isoelectric point and M(r) of proteins found in the transcriptome of the venom gland. The results also support the existence of posttranslational modifications and of proteolytic processing of precursor molecules which could lead to diverse multifunctional proteins. This study provides a preliminary reference map for proteins and peptides present in Bothrops insularis whole venom establishing the basis for comparative studies of other venom proteomes which could help the search for new drugs and the improvement of venom therapeutics. Altogether, our data point to the influence of transcriptional and post-translational events on the final venom composition and stress the need for a multivariate approach to snake venomics studies. (c) 2009 Elsevier B.V. All rights reserved.

Neuromuscular electrical stimulation improves GLUT-4 and morphological characteristics of skeletal muscle in rats with heart failure

Aim: Changes in skeletal muscle morphology and metabolism are associated with limited functional capacity in heart failure, which can be attenuated by neuromuscular electrical stimulation (ES). The purpose of the present study was to analyse the effects of ES upon GLUT-4 protein content, fibre structure and vessel density of the skeletal muscle in a rat model of HF subsequent to myocardial infarction. Methods: Forty-four male Wistar rats were assigned to one of four groups: sham (S), sham submitted to ES (S+ES), heart failure (HF) and heart failure submitted to ES (HF+ES). The rats in the ES groups were submitted to ES of the left leg during 20 days (2.5 kHz, once a day, 30 min, duty cycle 50%- 15 s contraction/15 s rest). After this period, the left tibialis anterior muscle was collected from all the rats for analysis. Results: HF+ES rats showed lower values of lung congestion when compared with HF rats (P = 0.0001). Although muscle weight was lower in HF rats than in the S group, thus indicating hypotrophy, 20 days of ES led to their recovery (P < 0.0001). In both groups submitted to ES, there was an increase in muscle vessel density (P < 0.04). Additionally, heart failure determined a 49% reduction in GLUT-4 protein content (P < 0.03), which was recovered by ES (P < 0.01). Conclusion: In heart failure, ES improves morphological changes and raises GLUT-4 content in skeletal muscle.

BnP1, a novel P-I metalloproteinase from Bothrops neuwiedi venom: Biological effects benchmarking relatively to jararhagin, a P-IIISVMP

Snake venom metalloproteinases (SVMPs) have been extensively studied and their effects associated with the local bleeding observed in human accidents by viper snakes. Representatives of P-I and P-III classes of SVMPs similarly hydrolyze extracellular matrix proteins or coagulation factors while only P-III SVMPs induce significant hemorrhage in experimental models. In this work, the effects of P-I and P-III SVMPs on plasma proteins and cultures of muscle and endothelial cells were compared in order to enlighten the mechanisms involved in venom-induced hemorrhage. To reach this comparison, BnP1 was isolated from B. neuwiedi venom and used as a weakly hemorrhagic P-I SVMPs and jararhagin was used as a model of potently hemorrhagic P-III SVMP. BnP1 was isolated by size exclusion and anion-exchange chromatographies, showing apparent molecular mass of approximately 24kDa and sequence similarity with other members of SVMPs, which allowed its classification as a group P-I SVMP. The comparison of local effects induced by SVMPs showed that BnP1 was devoid of significant myotoxic and hemorrhagic activities and jararhagin presented only hemorrhagic activity. BnP1 and jararhagin were able to hydrolyze fibrinogen and fibrin, although the latter displayed higher activity in both systems. Using HUVEC primary cultures, we observed that BnP1 induced cell detachment and a decrease in the number of viable endothelial cells in levels comparable to those observed by treatment with jararhagin. Moreover, both BnP1 and jararhagin induced apoptosis in HUVECs while only a small increase in LDH supernatant levels was observed after treatment with jararhagin, suggesting that the major mechanism involved in endothelial cell death is apoptosis. Jararhagin and BnP1 induced little effects on C2C12 muscle cell cultures, characterized by a partial detachment 24h after treatment and a mild necrotic effect as evidenced by a small increase in the supernatants LDH levels. Taken together, our data show that P-I and P-III SVMPs presented comparable effects except for the hemorrhagic activity, suggesting that hydrolysis of coagulation factors or damage to endothelial cells are not sufficient for induction of local bleeding. (C) 2007 Elsevier Ltd. All rights reserved.

Quality indices for (practical) clustering evaluation

Clustering quality or validation indices allow the evaluation of the quality of clustering in order to support the selection of a specific partition or clustering structure in its natural unsupervised environment, where the real solution is unknown or not available. In this paper, we investigate the use of quality indices mostly based on the concepts of clusters` compactness and separation, for the evaluation of clustering results (partitions in particular). This work intends to offer a general perspective regarding the appropriate use of quality indices for the purpose of clustering evaluation. After presenting some commonly used indices, as well as indices recently proposed in the literature, key issues regarding the practical use of quality indices are addressed. A general methodological approach is presented which considers the identification of appropriate indices thresholds. This general approach is compared with the simple use of quality indices for evaluating a clustering solution.

Association between particulate matter from biomass burning and respiratory diseases in the southern region of the Brazilian Amazon

Objective. To investigate the short-term effects of exposure to particulate matter from biomass burning in the Amazon on the daily demand for outpatient care due to respiratory diseases in children and the elderly. Methods. Epidemiologic study with ecologic time series design. Daily consultation records were obtained from the 14 primary health care clinics in the municipality of Alta Floresta, state of Mato Grosso, in the southern region of the Brazilian Amazon, between January 2004 and December 2005. Information on the daily levels of fine particulate matter was made available by the Brazilian National Institute for Spatial Research. To control for confounding factors ( situations in which a non-causal association between exposure and disease is observed due to a third variable), variables related to time trends, seasonality, temperature, relative humidity, rainfall, and calendar effects ( such as occurrence of holidays and weekends) were included in the model. Poisson regression with generalized additive models was used. Results. A 10 mu g/m(3) increase in the level of exposure to particulate matter was associated with increases of 2.9% and 2.6% in outpatient consultations due to respiratory diseases in children on the 6th and 7th days following exposure. Significant associations were not observed for elderly individuals. Conclusions. The results suggest that the levels of particulate matter from biomass burning in the Amazon are associated with adverse effects on the respiratory health of children.

Thermodynamic characterization of the palm tree Roystonea regia peroxidase stability

The structural stability of a peroxidase, a dimeric protein from royal palm tree (Roystonea regia) leaves, has been characterized by high-sensitivity differential scanning calorimetry, circular dichroism, steady-state tryptophan fluorescence and analytical ultracentifugation under different solvent conditions. It is shown that the thermal and chemical (using guanidine hydrochloride (Gdn-HCl)) folding/unfolding of royal palm tree peroxidase (RPTP) at pH 7 is a reversible process involving a highly cooperative transition between the folded dimer and unfolded monomers, with a free stabilization energy of about 23 kcal per mol of monomer at 25 degrees C. The structural stability of RPTP is pH-dependent. At pH 3, where ion pairs have disappeared due to protonation, the thermally induced denaturation of RPTP is irreversible and strongly dependent upon the scan rate, suggesting that this process is under kinetic control. Moreover, thermally induced transitions at this pH value are dependent on the protein concentration, allowing it to be concluded that in solution RPTP behaves as dimer, which undergoes thermal denaturation coupled with dissociation. Analysis of the kinetic parameters of RPTP denaturation at pH 3 was accomplished on the basis of the simple kinetic scheme N ->(k) D, where k is a first-order kinetic constant that changes with temperature, as given by the Arrhenius equation; N is the native state, and D is the denatured state, and thermodynamic information was obtained by extrapolation of the kinetic transition parameters to an infinite heating rate. Obtained in this way, the value of RPTP stability at 25 degrees C is ca. 8 kcal per mole of monomer lower than at pH 7. In all probability, this quantity reflects the contribution of ion pair interactions to the structural stability of RPTP. From a comparison of the stability of RPTP with other plant peroxidases it is proposed that one of the main factors responsible for the unusually high stability of RPTP which enhances its potential use for biotechnological purposes, is its dimerization. (c) 2008 Elsevier Masson SAS. All rights reserved.

Crystal structure and statistical coupling analysis of highly glycosylated peroxidase from royal palm tree (Roystonea regia)

Royal palm tree peroxidase (RPTP) is a very stable enzyme in regards to acidity, temperature, H(2)O(2), and organic solvents. Thus, RPTP is a promising candidate for developing H(2)O(2)-sensitive biosensors for diverse applications in industry and analytical chemistry. RPTP belongs to the family of class III secretory plant peroxidases, which include horseradish peroxidase isozyme C, soybean and peanut peroxidases. Here we report the X-ray structure of native RPTP isolated from royal palm tree (Roystonea regia) refined to a resolution of 1.85 angstrom. RPTP has the same overall folding pattern of the plant peroxidase superfamily, and it contains one heme group and two calcium-binding sites in similar locations. The three-dimensional structure of RPTP was solved for a hydroperoxide complex state, and it revealed a bound 2-(N-morpholino) ethanesulfonic acid molecule (MES) positioned at a putative substrate-binding secondary site. Nine N-glycosylation sites are clearly defined in the RPTP electron-density maps, revealing for the first time conformations of the glycan chains of this highly glycosylated enzyme. Furthermore, statistical coupling analysis (SCA) of the plant peroxidase superfamily was performed. This sequence-based method identified a set of evolutionarily conserved sites that mapped to regions surrounding the heme prosthetic group. The SCA matrix also predicted a set of energetically coupled residues that are involved in the maintenance of the structural folding of plant peroxidases. The combination of crystallographic data and SCA analysis provides information about the key structural elements that could contribute to explaining the unique stability of RPTP. (C) 2009 Elsevier Inc. All rights reserved.

Ausencia, prohibición y carencia : Estudio de los personajes y el deseo frustrado en tres obras de García Lorca

In this thesis, the main male characters in three of the plays written by Federico García Lorca are analysed with the aim of seeingthe role they play in the frustration of desire. After two chapters dedicated to a review of published critical studies on Lorca and tocertain theoretical considerations, Chapter Three examines desire drawing on Ubersfeld's actancial model and observes that thesemale characters can be divided into two groups: those who are desired and those who are undesired.In Chapter Four this classification is linked to an analysis of absence, prohibition and lack. Absence is here defined not asrelated to their non-appearance on stage but rather to their non-presence in the lives of the desiring female protagonists. It isobserved that a number of male characters are absent in the plays mainly due to death or a journey. As far as prohibition isconcerned, in two of the works, there is a moral code associated with concepts such as "honour" and "decency", which blocks thefemale characters' access to the males they desire. Chapter Four also shows how several characters can be considered as lacking inthe sense that they do not possess the ideal male qualities contained in the plays. This chapter reaches the conclusion that desiredmale characters are either absent or forbidden in the world of the desiring female, whereas undesired male characters are lacking inthe sense that they fail to live up to the ideal highlighted in the plays.Chapter Five analyses the female characters' perception of the male figures, making use of René Girard's notion of"transfiguration", which alludes to a process of idealisation of the object of desire. Our analysis reveals a connection betweendesire, denied access to the object of desire and transfiguration in the main subjects of desire. The phenomenon of "transfiguration"has several functions in the play: firstly, the creation of hyperbolical male characters; secondly, that of transmitting the intensity ofthe desire experienced and, finally, the highlighting of the lack of certain qualities in several male characters.We thus observe that, in these three plays written by García Lorca, Girard's pessimistic view of desire is confirmed, since desireneeds a series of obstacles, such as absence or prohibition, to survive. However, this is not the only explanation for the frustrationof desire: other factors, like the actions of certain male characters or destiny, also play a decisive role.

Fanfiction : la anonimicidad, el texto y el armario

This essay looks at slash, a genre within fanfiction, from the perspective of Sedgwick’s theory of the closet, which reflects on the concealing mechanisms associated with homosexuality. While the real author stays in the closet, disguised behind a pseudonym, slash texts present homosexual themes in a very explicit way, often relying on humor or subversive elements. Between these two spheres, the real author and the text, we can find what we call the author’s voice, conscious about the existing homophobic structures, a voice that uses different strategies to shield itself against them. Internet, with the possibility to stay anonymous, serves as a social closet where the masked authors create texts that subvert heteronormativity.

L’image comme outil didactique dans l’enseignement du français dans des collèges et des lycées en Suède

Le travail actuel a été rédigé dans le but d’éclaircir en quoi consiste le rôle de l’image comme outil didactique aux cours de français aux collèges et aux lycées suédois. Nous avons abordé notre thème sous des perspectives différentes : d’un côté, nous avons analysé les images dans deux livres, puisque les manuels font souvent partie de chaque leçon de français ; de l’autre côté, nous avons fait un questionnaire que nous avons distribué aux professeurs à travers les médias sociaux et par email. Nous avons également étudié les données récoltées aux cours des entretiens avec trois professeurs de français au lycée suédois, pour savoir comment les professeurs intègrent des images dans la pratique. Selon les résultats de notre étude, il nous a été possible d’affirmer que l’image est un outil qui n’est pas omniprésent, mais qui est parfois utilisé afin de rendre les leçons plus variées. La photographie est le type d’image le plus répandu dans la pratique des professeurs et dans les manuels ; le format numérique est le plus fréquent en tenant en compte de la révolution technologique et de l’habitude des jeunes à travailler avec les ordinateurs. Cependant, l’image n’est pas un outil indépendant. Elle est très souvent complétée par d’autres outils didactiques et sert principalement à accompagner la lecture, à développer l’expression orale et le vocabulaire et aussi à illustrer des phénomènes culturels. En conclusion, il ressort de notre mémoire, qu’il est important que le professeur accompagne le travail avec des images avec des instructions et des commentaires, afin de rendre l’apprentissage plus efficace.

Use of social network analysis to indentify the coopetition in a commercial cluster plan

The cluster provides a greater commercial relationship between the companies that comprise it. This encourages companies to adopt competitive structures that allow solving problems that would hardly alone (Lubeck et. Al., 2011). With that this paper aims to describe the coopetition between companies operating on a commercial cluster planned, from the point of view of retailers, taking as a basis the theoretical models proposed by Bengtsson and Kock (1999) and Leon (2005) and operationalized by means of Social Network Analysis (SNA). Data collection consisted of two phases, the first exploratory aspect to identify the actors, and the second was characterized as descriptive as it aims to describe the coopetition among the enterprises. As a result we identified the companies that cooperate and compete simultaneously (coopetition), firms that only compete, companies just cooperate and businesses that do not compete and do not cooperate (coexistence)

Estudo das propriedades psicométricas do perfil psicoeducacional PEP-R : elaboração da versão brasileira

O presente estudo investigou as propriedades psicométricas do Perfil Psicoeducacional Revisado (Psychoeducational Profile Revised - PEP-R), criado para avaliar a idade de desenvolvimento em crianças com autismo ou com transtornos correlatos da comunicação. Após tradução feita por dois tradutores bilíngües, vários itens (como imagens e letras do alfabeto) foram substituídos para tornar o material mais atrativo e apropriado à nossa cultura. Um estudo piloto composto por 20 crianças com desenvolvimento típico foi realizado para avaliar a adequação das mudanças realizadas nas imagens do instrumento. A validade relacionada a critério das dimensões da escala de Desenvolvimento e Comportamento foi avaliada administrando-se o PEP-R em 20 crianças autistas, 20 crianças com Síndrome de Down e 40 crianças com desenvolvimento típico. As crianças tinham idade entre 4 e 9 anos e também responderam ao teste de Matrizes Coloridas de Raven e à versão brasileira do CBCL (Child Behavior Checklist). As correlações entre o PEP-R e essas medidas foram de 0,54 e 0,39, respectivamente. A fidedignidade entre os avaliadores (coeficiente de W-Kendall) mostrou resultados variando entre 0,80 e 0,87. Os índices de consistência interna do PEP-R variaram entre 0,80 e 0,97. A comparação dos escores brutos finais da escala de Comportamento do PEP-R, nos três grupos, confirmou a validade discriminante do instrumento, uma vez que o grupo com autismo apresentou maior comprometimento que os demais grupos, nas dimensões investigadas

Monetary arrangements for emerging economies

In this paper we look at various alternatives for monetary regimes: dollarization, monetary union and local currency. We use an extension of the debt crisis model of Cole and Kehoe ([3], [4] and [5]), although we do not necessarily follow their sunspot interpretation. Our focus is to appraise the welfare of a country which is heavily dependent on international capital due to low savings, for example, and might suffer a speculative attack on its external public debt. We study the conditions under which countries will be better off adopting each one of the regimes described above. If it belongs to a monetary union or to a local currency regime, a default may be avoided by an ination tax on debt denominated in common or local currency, respectively. Under the former regime, the decision to inate depends on each member country's political inuence over the union's central bank, while, in the latter one, the country has full autonomy to decide about its monetary policy. The possibility that the government inuences the central bank to create ination tax for political reasons adversely affects the expected welfare of both regimes. Under dollarization, ination is ruled out and the country that is subject to an external debt crisis has no other option than to default. Accordingly, one of our main results is that shared ination control strengthens currencies and a common-currency regime is superior in terms of expected welfare to the local-currency one and to dollarization if external shocks that member countries suffer are strongly correlated to each other. On the other hand, dollarization is dominant if the room for political ination under the alternative regime is high. Finally, local currency is dominant if external shocks are uncorrelated and the room for political pressure is mild. We nish by comparing Brazil's and Argentina's recent experiences which resemble the dollarization and the local currency regimes, and appraising the incentives that member countries would have to unify their currencies in the following common markets: Southern Common Market, Andean Community of Nations and Central American Common Market.

Speculative attacks on debts and optimum currency area: a welfare analysis

Traditionally the issue of an optimum currency area is based on the theoretical underpinnings developed in the 1960s by McKinnon [13], Kenen [12] and mainly Mundell [14], who is concerned with the benefits of lowering transaction costs vis-à- vis adjustments to asymmetrical shocks. Recently, this theme has been reappraised with new aspects included in the analysis, such as: incomplete markets, credibility of monetary policy and seigniorage, among others. For instance, Neumeyer [15] develops a general equilibrium model with incomplete asset markets and shows that a monetary union is desirable when the welfare gains of eliminating the exchange rate volatility are greater than the cost of reducing the number of currencies to hedge against risks. In this paper, we also resort to a general equilibrium model to evaluate financial aspects of an optimum currency area. Our focus is to appraise the welfare of a country heavily dependent on foreign capital that may suffer a speculative attack on its public debt. The welfare analysis uses as reference the self-fulfilling debt crisis model of Cole and Kehoe ([6], [7] and [8]), which is employed here to represent dollarization. Under this regime, the national government has no control over its monetary policy, the total public debt is denominated in dollars and it is in the hands of international bankers. To describe a country that is a member of a currency union, we modify the original Cole-Kehoe model by including public debt denominated in common currency, only purchased by national consumers. According to this rule, the member countries regain some influence over the monetary policy decision, which is, however, dependent on majority voting. We show that for specific levels of dollar debt, to create inflation tax on common-currency debt in order to avoid an external default is more desirable than to suspend its payment, which is the only choice available for a dollarized economy when foreign creditors decide not to renew their loans.

Speculative attacks, openness and crises

In this paper we propose a dynamic stochastic general equilibrium model to evaluate financial adjustments that some emerging market economies went through to overcome external crises during the latest decades, such as default and local currency devaluation. We assume that real devaluation can be used to avoid external debt default, to improve trade balance and to reduce the real public debt level denominated in local currency. Such effects increase the government ability to deal with external crisis, but also have costs in terms of welfare, related to expected inflation, reductions in private investments and higher interest to be paid over the public debt. We conclude that openness improves expected welfare as it allows for a better devaluation-response technology against crises. We also present results for 32 middle-income countries, verifying that the proposed model can indicate, in a stylized way, the preferences for default-devaluation options and the magnitude of the currency depreciation required to overcome 48 external crises occurred as from 1971. Finally, as we construct our model based on the Cole-Kehoe self-fulfilling debt crisis model ([7]), adding local debt and trade, it is important to say that their policy alternatives to leave the crisis zone remains in our extended model, namely, to reduce the external debt level and to lengthen its maturity.