Purificação, caracterização e avaliação da atividade antiproliferativa de um inibidor de quimotripsina tipo kunitz de semente de Eryhrina velutina

A chymotrypsin inhibitor was purified from Erythrina velutina seeds by ammonium sulphate fractionation, affinities chromatographies on Trypsin-Sepharose, Quimotrypsin-Sepharose and reversed phase C-18 FPLC/AKTA system. The inhibitor, named EvCI, shown molecular mass of 17 kDa, as determined by SDSPAGE. 2D-PAGE showed four isoinhibitors with pI values of 4,42, 4,63, 4,83 and 5,06, with molecular mass of 17 kDa each. The aminoacid sequence of EvCI was determined by MALDI-TOF-MS and showed a high similarity with other Kunitz-type inhibitor of Erythrina variegata. EvCI competitively inhibited chymotrypsin, with Ki of 4 x10-8 M, but did not inhibited trypsin, pancreatic elastase, bromelain and papain. The inhibitory activity of EvCI was stable over wide pH and temperature ranges. In the presence of DTT 100 mM for 120 min, EvCI lost 50 % of activity. Cytotoxicity was studied in HeLa, MDA, HepG2, K562 and PC3 cells after 72-h incubation period. EvCl inhibited HeLa cells growth with an IC50 value of 50 μg/ml. Subsequent studies in HeLa cells analysis of cell death by annexin V/PI double-staining and cell cycle, using flow cytometry. The results provide evidence for a cytostatic activity of EvCl and support further studies on potential application of this inhibitors as an antiproliferative agent in combined therapy against cervical cancer

Ação Sacietogênica de um inibidor de tripsina da semente de tamarindo (Tamarindus indica L.)

The seeds are excellent sources of proteinase inhibitors and have been highlighted owing to various applications. Among these applications are those in effect on food intake and weight gain that stand out because of the increasing number of obese individuals. This study evaluated the effects of trypsin inhibitor present in the seed of tamarind (Tamarindus indica L.) reduction in weight gain, biochemical and morphological alterations in Wistar rats. For this, we partially purified a trypsin inhibitor tamarind seed. This inhibitor, ITT2 at a concentration of 25 mg / kg body weight, over a period of 14 days was able to reduce food intake in rats (n = 6) by approximately 47%, causing a reduction in weight gain approximately 70% when compared with the control group. With the evaluation of the in vivo digestibility was demonstrated that the animals lost weight due to satiety, presented by the reduction of food intake, since there were significant differences between true digestibility for the control group (90.7%) and the group treated with inhibitor (89.88%). Additionally, we checked the deeds of ITT2 on biochemical parameters (glucose, triglycerides, total cholesterol, high-density lipoprotein, low-density lipoprotein, glutamic-pyruvic transaminase, glutamic oxaloacetic transaminase, gamma glutamyl transferase albumin, globulin, total protein and C-reactive protein) and these, when assessed in the study groups showed no statistically significant variations. We also evaluate the histology of some organs, liver, stomach, intestine, and pancreas, and showed no changes. And to evaluate the effect of trypsin inhibitor on food intake due to the satiety is regulated by cholecystokinin (CCK) were measured plasma levels, and it was observed that the levels of CCK in animals receiving ITT2 were significantly higher ( 20 + 1.22) than in animals receiving only solution with casein (10.14 + 2.9) or water (5.92 + 1.15). Thus, the results indicate that the effect caused ITT2 satiety, reducing food intake, which in turn caused a reduction in weight gain in animals without causing morphological and biochemical changes, this effect caused by the elevation of plasma levels CCK

Mobilização de reservas e partição de metabólitos durante a germinação da semente e o estabelecimento da plântula em moringa

Seed germination and seedling establishment are critical processes for commercial plantation and depend directly on reserve mobilization as a source of cellular fuels and biosynthetic precursors. In this way, we investigated the coordination among reserve mobilization, metabolite partitioning, and mobilizing enzyme activities in Moringa oleifera Lam (moringa) an oil-seeded species employed in biofuel production. Seeds were germinated under controlled conditions and seedlings were grown hydroponically at a greenhouse. Samples were harvested at 0, 4, 8, 10, 12, 16, and 20 days after imbibition (DAI). The contents of dry mass (DM), neutral lipids (NL), soluble proteins (SP), starch, total soluble sugars (TSS), non-reducing sugars (NRS), and total free amino acids (TFAA) as the activity of isocitrate lyase (ICL), acid proteases, and amylases were determined. The mobilization of storage proteins was initiated during seed germination whereas the mobilization of storage lipids and starch was triggered throughout seedling establishment although all reserves have been depleted until 20 DAI. The partitioning of DM and metabolites to the roots and the shoots was uneven during seedling establishment. Low shoot/root ratio on the basis of DM could be related to the natural occurrence of moringa in drought climates. In the roots, TSS, NRS, and TFAA were accumulated from 12 to 16 DAI and then were consumed until the end of the experiment. In the shoots, TSS and TFAA were consumed in parallel with NRS accumulation from 12 to 20 DAI. The activity of ICL, acid proteases, and amylases was coordinated with the mobilization of lipids, proteins and starch respectively. Thus, we propose that the patterns of reserve mobilization and metabolite partitioning verified in moringa seem distinct from those found to other tree species and may be involved in metabolic strategies to enable environment colonization

Programa nova semente em perspectiva (2009-2012): uma avaliação de processo

Historically the provision of childcare facilities in Brazil was short of demand. This problem affects a large part of the population and has a tendency to worsen, due to the accelerated decline in the number of parents of dedication exclusive family. The program called "Nova Semente" seeks to accelerate the creation of childcare facilities in the county, enabling the opening of units at low cost and in record time, through partnerships between government, non-governmental and civil society organizations. This study conducted a process evaluation of the implementation of public policy, to identify the strengths and weaknesses of the policy. In real terms, this paper seeks to understand the motivation for the development of the program, identifying the reasons set out in the project match what is being sent; Understand how it is giving practical partnership between the government entity and the entities "non-state"; Identify the degree of achievement of goals that should be met even in the implementation phase; verify employment of instruments provided for its implementation. For this, the literature of public policy evaluation is used, and fits the criteria and mechanisms for analysis in the assessment of efficacy, to answer the question motivating the work, which deals with uncertainty about the durability of the Program. the framework is the criteria and mechanisms for analysis in the assessment of efficacy, to answer the question motivating the work, which deals with uncertainty about the durability of the Program

"Eis que a semente caiu em boa terra": a avenida Cruz Cabugá no Recife-PE como campo religioso local

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Equilíbrio líquido-líquido para o sistema biodiesel de óleo de semente de melão (Cucumis melo l.) + metanol + glicerina

Rio Grande do Norte, northeast state from Brazil, it is the greatest producer and exporter of yellow melon, well known as Spanish melon. Despite the consumption of this fruit to be mainly its pulp, melon seeds are an important source of lipids considered an industrial residue it has been discharge product. The use of oilseeds in order to produce biodiesel establishes an important raw material and the increase of its production promotes the national development of the agriculture. In this background, the aim of this work has been to use oil from seeds of yellow melon to produce biodiesel and to accomplish a study of the phase equilibrium of the system evolving biodiesel, methanol and glycerin. The biodiesel was obtained by oil transesterification through methylic route with molar ratio 1:9.7 (oil:alcohol) and with a mass of NaOH of 0.5% from the oil mass; the reaction time was 73 minutes at 55 °C. A yield of 84.94% in biodiesel was achieved. The equilibria data present a well-characterized behavior with a great region of two phases. The tie lines indicate that methanol has a best solubility in the phase that is rich in glycerin. Consistency of the experimental data was made based on Othmer-Tobias and Hand correlations which values above 0.99 were found to correlation coefficients, this fact confers a good thermodynamic consistency to the experimental data. NRTL and UNIQUAC models were employed to predict liquid-liquid equilibrium of this system. It was observed a better concordance of the results when NRTL was applied (standard deviation 1.25%) although the UNIQUAC model has presented a quite satisfactory result either (standard deviation 2.70%). The NRTL and UNIQUAC models were also used to evaluate the effect of temperature in the range of 328 K to 358 K, in which a little change in solubility with respect to the data obtained at 298 K was observed, thus being considered negligible the effect of temperature

Estudo da circulação hidrodinâmica e da dispersão de óleo na zona costeira entre Macau e Galinhos RN

This paper presents a study on coastal hydrodynamics and the spread of an oil spill in waters off Macau and Galinhos, on the east coast of the state of Rio Grande do Norte in Northeast Brazil. This area has a very marked coastal dynamic owing to the complexity of its geomorphological features, developed in a regime of semidiurnal mesotides involving reefs, spits, estuaries, mangroves, lakes and dunes. The region also plays an important role in the socioeconomic development of the state, given that the production of oil, natural gas, salt and shrimp is concentrated there. The series of oil platforms is interconnected by a pipeline system that carries oil to the local terminal. This pipeline could leak at any moment, causing immense ecological damage. To gauge the risks of an oil leak and resulting contamination of the coastal region, two hydrodynamic scenarios were simulated. The results obtained were used to implement a contaminant transport model with the creation of various oil leak scenarios modeled at different volumes (from small to large) and intensities (sporadic and continuous), at points considered critical for the model (on two platforms and at two pipeline intersections), under different wind (summer and winter) and tidal (high and low at new, full and quarter moon phases) conditions. The use of hydrodynamic circulation computer models as a tool for representing a real project design has been increasingly frequent in recent years, given that they enable the realistic simulation of the hydrodynamic circulation pattern in bodies of water and an analysis of the impacts caused by contaminants released into the water. This study used the computer models contained in SisBAHIA®, in continuous development in the area of Coastal Engineering and Oceanography at COPPE/UFRJ

Simulações da ação do vento e da dispersão de contaminantes no estuário do Rio Potengi/ Brasil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Sair curado para a vida e para o bem: diagramas, linhas e dispersão de forças no complexus nosoespacial do Hospital de Caridade Juvino Barreto (1909-1927)

This work has as object of study the Hospital de Caridade Juvino Barreto, nosocomial institution located in the city of Natal (RN), between the Praia de Areia Preta and the Monte Petrópolis, focusing on the period from 1909, the year in which the new hospital building was constructed and opened, and 1927, the date of the transfer of administration of the public domain to the newly created Sociedade de Assistência Hospitalar (SAH). We study the conditions of possibility of the emergence of this hospital space in the urban environment of the capital of Rio Grande do Norte, seeking to understand the different tactics and strategies implemented by the historical subjects involved in the formation of this institution nosocomial. Starting from a corpus of documents consisting of medical memories (with Dr. Januário Cicco as privileged observer), information present in newspapers (the Republic and the Christmas Journa l), photo collection and extensive administrative and legal material (Speeches, Exhibitions, Reports, Laws and Resolutions), we analyzed in detail the medical geography of HCJB, relating the discourses of medicine and geography in choosing the spatial location of the hospital as we examine the architecture of the hospital, its inner spat iality, divisions, forms of space control, and, finally, we discuss the medical practices that took place within it, leading us in this regard, from the experiences of clinical hospital chief, Dr. Januário Cicco, especially the discussion on "ethics" in hospital work. The perception of HCJB as medical nosoespaciality always on the move, incorporated under taxonomic principles based on difference and dispersion forces, led us to articulate it theoretically from the conceptual-methodological arsenal of philosopher Michel Foucault, particularly his reflections of genealogical phase, focusing on the phenomenon of power, a position that allows us to enhance our space-hospital construction, invention, product of power relations, which give the unfinished aspect nosocômio, apparent, always at stake, perpetual non-modeling possibility has previously defined array, establishing it at the field of possible, of virtuality, of power: hospital that could have been and that it was not. Indeed, the investigation of various aspects/elements of hospital space Juvino Barreto revealed us new dimensions of hospital space, far more complex than the simple and the current idea of a place to shelter patients: plasticity and fluidity of space, which is not made to circumscribe the limits of empeiria, engraving up to strength relations fought between different subject; its Constitution as a transitional space, Heterotopic, doing live inside modern elements with premoderns (professional doctors working with religious thought, skeptical of positivist medicine living with the religious faith of the nuns of Santana); the impossibility of thinking hospital space of HCJB while homogeneous unit, static, transistoric, making the spatiality, without considering the profound differences, fractures and dislocations that animated his own existence, multiplying their expressions of identity

Modelos de dispersão para extremos de precipitação. Estudo de caso: o nordeste do Brasil

In this thesis used four different methods in order to diagnose the precipitation extremes on Northeastern Brazil (NEB): Generalized Linear Model s via logistic regression and Poisson, extreme value theory analysis via generalized extre me value (GEV) and generalized Pareto (GPD) distributions and Vectorial Generalized Linea r Models via GEV (MVLG GEV). The logistic regression and Poisson models were used to identify the interactions between the precipitation extremes and other variables based on the odds ratios and relative risks. It was found that the outgoing longwave radiation was the indicator variable for the occurrence of extreme precipitation on eastern, northern and semi arid NEB, and the relative humidity was verified on southern NEB. The GEV and GPD distribut ions (based on the 95th percentile) showed that the location and scale parameters were presented the maximum on the eastern and northern coast NEB, the GEV verified a maximum core on western of Pernambuco influenced by weather systems and topography. The GEV and GPD shape parameter, for most regions the data fitted by Weibull negative an d Beta distributions (ξ < 0) , respectively. The levels and return periods of GEV (GPD) on north ern Maranhão (centerrn of Bahia) may occur at least an extreme precipitation event excee ding over of 160.9 mm /day (192.3 mm / day) on next 30 years. The MVLG GEV model found tha t the zonal and meridional wind components, evaporation and Atlantic and Pacific se a surface temperature boost the precipitation extremes. The GEV parameters show the following results: a) location ( ), the highest value was 88.26 ± 6.42 mm on northern Maran hão; b) scale ( σ ), most regions showed positive values, except on southern of Maranhão; an d c) shape ( ξ ), most of the selected regions were adjusted by the Weibull negative distr ibution ( ξ < 0 ). The southern Maranhão and southern Bahia have greater accuracy. The level period, it was estimated that the centern of Bahia may occur at least an extreme precipitatio n event equal to or exceeding over 571.2 mm/day on next 30 years.

Investigando o uso do sentido háptico na visualização de dados multivariados em gráficos de dispersão bidimensionais

Data Visualization is widely used to facilitate the comprehension of information and find relationships between data. One of the most widely used techniques for multivariate data (4 or more variables) visualization is the 2D scatterplot. This technique associates each data item to a visual mark in the following way: two variables are mapped to Cartesian coordinates so that a visual mark can be placed on the Cartesian plane; the others variables are mapped gradually to visual properties of the mark, such as size, color, shape, among others. As the number of variables to be visualized increases, the amount of visual properties associated to the mark increases as well. As a result, the complexity of the final visualization is higher. However, increasing the complexity of the visualization does not necessarily implies a better visualization and, sometimes, it provides an inverse situation, producing a visually polluted and confusing visualization—this problem is called visual properties overload. This work aims to investigate whether it is possible to work around the overload of the visual channel and improve insight about multivariate data visualized through a modification in the 2D scatterplot technique. In this modification, we map the variables from data items to multisensoriy marks. These marks are composed not only by visual properties, but haptic properties, such as vibration, viscosity and elastic resistance, as well. We believed that this approach could ease the insight process, through the transposition of properties from the visual channel to the haptic channel. The hypothesis was verified through experiments, in which we have analyzed (a) the accuracy of the answers; (b) response time; and (c) the grade of personal satisfaction with the proposed approach. However, the hypothesis was not validated. The results suggest that there is an equivalence between the investigated visual and haptic properties in all analyzed aspects, though in strictly numeric terms the multisensory visualization achieved better results in response time and personal satisfaction.

Estrutura crustal e mantélica da província Borborema através de funções do receptor e dispersão de ondas superficiais

The Borborema Province, located in northeastern Brazil, has a basement of Precambrian age and a tectonic framework structured at the Neoproterozoic (740-560 Ma). After separation between South America and Africa during the Mesozoic, a rift system was formed, giving rise to a number of marginal and inland basins in the Province. After continental breakup, episodes of volcanism and uplift characterized the evolution of the Province. Plateau uplift was initially related to magmatic underplating of mafic material at the base of the crust, perhaps related to the generation of young continental plugs (45-7 Ma) along the Macau-Queimadas Alignment (MQA), due to a small-scale convection at the continental edge. The goal of this study is to investigate the causes of intra-plate uplift and its relationship to MQA volcanism, by using broadband seismology and integrating our results with independent geophysical and geological studies in the Borborema Province. The investigation of the deep structure of the Province with broadband seismic data includes receiver functions and surface-wave dispersion tomography. Both the receiver functions and surface-wave dispersion tomography are methods that use teleseismic events and allow to develop estimates of crustal parameters such as crustal thickness, Vp/Vs ratio, and S-velocity structure. The seismograms used for the receiver function work were obtained from 52 stations in Northeast Brazil: 16 broadband stations from the RSISNE network (Rede Sismográfica do Nordeste do Brasil), and 21 short-period and 6 broadband stations from the INCT-ET network (Instituto Nacional de Ciência e Tecnologia – Estudos Tectônicos). These results add signifi- cantly to previous datasets collected at individual stations in the Province, which include station RCBR (GSN - Global Seismic Network), stations CAUB and AGBL (Brazilian Lithosphere Seismic Project IAG/USP), and 6 other broadband stations that were part of the Projeto Milênio - Estudos geofísicos e tectônicos na Província Borborema/CNPq. For the surface-wave vii tomography, seismograms recorde at 22 broadband stations were utilized: 16 broadband stations from the RSISNE network and 6 broadband stations from the Milênio project. The new constraints developed in this work include: (i) estimates of crustal thickness and bulk Vp/Vs ratio for each station using receiver functions; (ii) new measurements of surfassewave group velocity, which were integrated to existing measurementes from a continental-scale tomography for South America, and (iii) S-wave velocity models (1D) at various locations in the Borborema Province, developed through the simultaneous inversion of receiver functions and surface-wave dispersion velocities. The results display S-wave velocity structure down to the base of the crust that are consistent with the presence of a 5-7.5 km thick mafic layer. The mafic layer was observed only in the southern portion of the Plateau and absent in its northern portion. Another important observation is that our models divide the plateau into a region of thin crust (northern Plateau) and a region of thick crust (southern Plateau), confirming results from independent refraction surveys and receiver function analyses. Existing models of plateau uplift, nonetheless, cannot explain all the new observations. It is proposed that during the Brazilian orogeny a layer of preexisting mafic material was delaminated, as a whole or in part, from the original Brasiliano crust. Partial delamination would have happened in the southern portion of the plateau, where independent studies found evidence of a more resistant rheology. During Mesozoic rifting, thinning of the crust around the southern Plateau would have formed the marginal basins and the Sertaneja depression, which would have included the northern part of the Plateau. In the Cenozoic, uplift of the northern Plateau would have occurred, resulting in a northern Plateau without mafic material at the base of the crust and a southern Plateau with partially delaminated mafic layer.

Purificação, caracterização e avaliação da atividade antiproliferativa de um inibidor de quimotripsina tipo kunitz de semente de Eryhrina velutina

A chymotrypsin inhibitor was purified from Erythrina velutina seeds by ammonium sulphate fractionation, affinities chromatographies on Trypsin-Sepharose, Quimotrypsin-Sepharose and reversed phase C-18 FPLC/AKTA system. The inhibitor, named EvCI, shown molecular mass of 17 kDa, as determined by SDSPAGE. 2D-PAGE showed four isoinhibitors with pI values of 4,42, 4,63, 4,83 and 5,06, with molecular mass of 17 kDa each. The aminoacid sequence of EvCI was determined by MALDI-TOF-MS and showed a high similarity with other Kunitz-type inhibitor of Erythrina variegata. EvCI competitively inhibited chymotrypsin, with Ki of 4 x10-8 M, but did not inhibited trypsin, pancreatic elastase, bromelain and papain. The inhibitory activity of EvCI was stable over wide pH and temperature ranges. In the presence of DTT 100 mM for 120 min, EvCI lost 50 % of activity. Cytotoxicity was studied in HeLa, MDA, HepG2, K562 and PC3 cells after 72-h incubation period. EvCl inhibited HeLa cells growth with an IC50 value of 50 μg/ml. Subsequent studies in HeLa cells analysis of cell death by annexin V/PI double-staining and cell cycle, using flow cytometry. The results provide evidence for a cytostatic activity of EvCl and support further studies on potential application of this inhibitors as an antiproliferative agent in combined therapy against cervical cancer

Ação Sacietogênica de um inibidor de tripsina da semente de tamarindo (Tamarindus indica L.)

The seeds are excellent sources of proteinase inhibitors and have been highlighted owing to various applications. Among these applications are those in effect on food intake and weight gain that stand out because of the increasing number of obese individuals. This study evaluated the effects of trypsin inhibitor present in the seed of tamarind (Tamarindus indica L.) reduction in weight gain, biochemical and morphological alterations in Wistar rats. For this, we partially purified a trypsin inhibitor tamarind seed. This inhibitor, ITT2 at a concentration of 25 mg / kg body weight, over a period of 14 days was able to reduce food intake in rats (n = 6) by approximately 47%, causing a reduction in weight gain approximately 70% when compared with the control group. With the evaluation of the in vivo digestibility was demonstrated that the animals lost weight due to satiety, presented by the reduction of food intake, since there were significant differences between true digestibility for the control group (90.7%) and the group treated with inhibitor (89.88%). Additionally, we checked the deeds of ITT2 on biochemical parameters (glucose, triglycerides, total cholesterol, high-density lipoprotein, low-density lipoprotein, glutamic-pyruvic transaminase, glutamic oxaloacetic transaminase, gamma glutamyl transferase albumin, globulin, total protein and C-reactive protein) and these, when assessed in the study groups showed no statistically significant variations. We also evaluate the histology of some organs, liver, stomach, intestine, and pancreas, and showed no changes. And to evaluate the effect of trypsin inhibitor on food intake due to the satiety is regulated by cholecystokinin (CCK) were measured plasma levels, and it was observed that the levels of CCK in animals receiving ITT2 were significantly higher ( 20 + 1.22) than in animals receiving only solution with casein (10.14 + 2.9) or water (5.92 + 1.15). Thus, the results indicate that the effect caused ITT2 satiety, reducing food intake, which in turn caused a reduction in weight gain in animals without causing morphological and biochemical changes, this effect caused by the elevation of plasma levels CCK

Construção de modelos de interação in silico e in vitro do inibidor do tipo Kunitz de Adenanthera pavonina L. para as enzimas cisteínicas e serínicas

Serines proteinases inhibitors (PIs) are widely distributed in nature and are able to inhibit both in vitro and in vivo enzymatic activites. Seed PIs in than leguminous are classified in seven families, Bowman-Birk and Kunitz type families that most studied representing an important role in the first line of defense toward insects pests. Some Kunitz type inhibitors possess activities serine and cysteine for proteinases named bifunctional inhibitor, as ApTKI the inhibitor isolate from seed of Adenanthera pavonina. The A. pavonina inhibitor presenting the uncommon property and was used for interaction studies between proteinases serine (trypsin) and cysteine (papain). In order to determinate the in vitro interaction of ApTKI against enzymes inhibitor purification was carried cut by using chromatographic techniques and inhibition assays. The 3D model of the bifunctional inhibitor ApTKI was constructed SWISS-MODEL program by homology modeling using soybean trypsin inhibitor (STI, pdb:1ba7), as template which presented 40% of identity to A. pavonina inhibitor. Model quality was evaluated by PROCHECK program. Moreover in silico analyzes of formed complex between the enzymes and ApTKI was evaluated by HEX 4.5 program. In vitro results confirmed the inhibitory assays, where the inhibitor presented the ability to simultaneously inhibit trypsin and papain. The residues encountered in the inhibitor model of folder structural three-dimensional that make contact to enzymes target coud explain the specificity pattern against serine and cysteine proteinases