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

Purificação, caracterização e efeitos imunomodulatório e antiproliferativo de uma lectina do fungo Clavaria cristata (Holmsk.) Pers

A 140,0 kDa lectin was purified and characterized from the mushroom Clavaria cristata. The purification procedures from the crude extract of the mushroom comprised gel filtration chromatography on Sephacryl s200 and ion exchange on Resource Q column. The purified lectin agglutinated all types of human erythrocytes with preference for trypsinized type O erythrocytes. The haemagglutinating activity is dependent of Ca 2+ ions and was strongly inhibited by the glycoprotein bovine submaxillary mucin (BSM) up to the concentration of 0, 125 mg/mL. The C. cristata lectin (CcL) was stable in the pH range of 2,5-11,5 and termostable up to 80 °C. CcL molecular mass determined by gel filtration on a Superose 6 10 300 column was approximately 140,3 kDa. SDS polyacrilamide gel electrophoresis revealed a single band with a molecular mass of approximately 14,5 kDa, when the lectin was heated at 100 ⁰C in the presence or absence of β-mercaptoethanol. CcL induced activation of murine peritoneal macrophages in vitro resulting in the release of nitric oxide (NO), reaching the maximum production at 24 h. In experimental paw oedema model in mice, CcL showed proinflammatory activity being able to induce oedema formation. Cell viability of HepG2, MDA 435 e 3T3 cell lines was examined after 72 h of incubation with CcL in different concentrations (0,5-50 μg/mL). CcL inhibited HepG2 cells growth with an IC50 value of 50 μg/mL. In the present work, the observed immunomodulatory and antiproliferative effects indicate CcL as a possible immunomodulator compound, interfering in the macrophages immune response, taking possible anti-parasitic, anti-tumoral effects or diagnostic and/or therapeutic

Políticas públicas para agricultura familiar: uma avaliação sobre a correspondência do paa e do PRONAF nos territórios da cidadania do Rio Grande do Norte - 2008 a 2010.

Family farming has been considered as the new axis of rural development in the country, the focus of several public policies, especially the National Program for Strengthening Family Agriculture - PRONAF and Food Purchase Program - PAA. PRONAF was created with the aim of providing credit to farmers, while the PAA to support family farmers through the purchase of its production. In this context, the objective of this study is to analyze the correspondence of these two public policies for family farming, in the Territories of Citizenship of the state of Rio Grande do Norte, between the years 2008 to 2010. In the methodology, the analysis was performed by comparing the distributions of the two programs in the territories of citizenship status. There were also statistical tests of differences in proportions, and Spearman correlations, and estimated a logit regression model, in order to measure the probability of a farmer participating in the PAA is associated with one of the modes of PRONAF. The data used were obtained from the National and Supply - CONAB at the Institute of Technical Assistance and Rural Extension - EMATER, and the Ministry of Agrarian Development - MDA. Among the key findings was noted that policies were associated with a direct, but low in the districts of the Territories of Citizenship. And that, in the years 2008 and 2009, only in the territories of Mato Grande, Alto Oeste and Seridó, the actions of PAA and PRONAF had direct and significant correlations. It was found that in most of the territories, policies are performed randomly, ie that both have no correlation to each other. The estimates of the logit model showed that the chance of a family farmer, the PAA participant, receive credits PRONAF A, is higher in the territory of Mato Grande, and would have a chance to fall in PRONAF B in all areas surveyed. Moreover, farmers in the territories of the Assu-Mossoró, Sertão of Apodi, Seridó and Alto Oeste, participating in the PAA would be more likely to receive credits PRONAF C, reflecting thus the family farm more consolidated these territories

Uma linguagem de modelagem da interação para auxiliar a comunicação designer-usuário

The development of interactive systems involves several professionals and the integration between them normally uses common artifacts, such as models, that drive the development process. In the model-driven development approach, the interaction model is an artifact that includes the most of the aspects related to what and how the user can do while he/she interacting with the system. Furthermore, the interactive model may be used to identify usability problems at design time. Therefore, the central problematic addressed by this thesis is twofold. In the first place, the interaction modeling, in a perspective that helps the designer to explicit to developer, who will implement the interface, the aspcts related to the interaction process. In the second place, the anticipated identification of usability problems, that aims to reduce the application final costs. To achieve these goals, this work presents (i) the ALaDIM language, that aims to help the designer on the conception, representation and validation of his interactive message models; (ii) the ALaDIM editor, which was built using the EMF (Eclipse Modeling Framework) and its standardized technologies by OMG (Object Management Group); and (iii) the ALaDIM inspection method, which allows the anticipated identification of usability problems using ALaDIM models. ALaDIM language and editor were respectively specified and implemented using the OMG standards and they can be used in MDA (Model Driven Architecture) activities. Beyond that, we evaluated both ALaDIM language and editor using a CDN (Cognitive Dimensions of Notations) analysis. Finally, this work reports an experiment that validated the ALaDIM inspection method

Uma abordagem para promover o alinhamento entre a estratégia de negócio e a tecnologia de informação

Currently with the increase in complexity in doing business, organizations are seeking information systems that help to quickly respond to new demands in the processes of production of products and services. An information system is no longer just a support tool and has become an integral part of doing business. However, in spite of significant technological evolution in recent years, information systems that support business do not respond efficiently to the constant alterations that occur in many organizations. One of the main problems faced by information systems currently is the lack of strategic alignment between business strategy and information technology. The concept of strategic alignment can be defined as a way between business strategies and objectives and the strategies, objectives and functions of information technology in such as way as to contribute to the increase in competitivity of the organization over time. Strategic alignment together with strategic planning are important management instruments. Approaches for operationalizing this alignment are being developed currently but are still in their initial stages due to the fact that it is a relatively new concept in the literature. Another point that needs to be taken into consideration during the strategic alignment is the question of trackability between the business elements and IT. Trackability (Tracking) is necessary for example when one wishes to know exactly which goal defined in the business strategy was left out or not accepted due to a modification made in the IT strategy. Very few proposals present concrete ways supported by software systems in order to obtain strategic alignement while taking into consideration this trackability. Therefore the objective of this work is to propose the creation of a strategic alignment process supported by a software system which is capable of permitting trackability between the organizational objectives and the business processes based on formalization standards defined through a model oriented approach

CrossMDA2: Uma abordagem para minimizar o problema da fragilidade de pointcuts na evolução de sistemas orientados a aspectos

This work proposes a model based approach for pointcut management in the presence of evolution in aspect oriented systems. The proposed approach, called conceptual visions based pointcuts, is motivated by the observation of the shortcomings in traditional approaches pointcuts definition, which generally refer directly to software structure and/or behavior, thereby creating a strong coupling between pointcut definition and the base code. This coupling causes the problem known as pointcut fragility problem and hinders the evolution of aspect-oriented systems. This problem occurs when all the pointcuts of each aspect should be reviewed due to any software changes/evolution, to ensure that they remain valid even after the changes made in the software. Our approach is focused on the pointcuts definition based on a conceptual model, which has definitions of the system's structure in a more abstract level. The conceptual model consists of classifications (called conceptual views) on entities of the business model elements based on common characteristics, and relationships between these views. Thus the pointcuts definitions are created based on the conceptual model rather than directly referencing the base model. Moreover, the conceptual model contains a set of relationships that allows it to be automatically verified if the classifications in the conceptual model remain valid even after a software change. To this end, all the development using the conceptual views based pointcuts approach is supported by a conceptual framework called CrossMDA2 and a development process based on MDA, both also proposed in this work. As proof of concept, we present two versions of a case study, setting up a scenario of evolution that shows how the use of conceptual visions based pointcuts helps detecting and minimizing the pointcuts fragility. For the proposal evaluation the Goal/Question/Metric (GQM) technique is used together with metrics for efficiency analysis in the pointcuts definition

Abordagem dirigida a modelos para redes de sensores sem fio

Research on Wireless Sensor Networks (WSN) has evolved, with potential applications in several domains. However, the building of WSN applications is hampered by the need of programming in low-level abstractions provided by sensor OS and of specific knowledge about each application domain and each sensor platform. We propose a MDA approach do develop WSN applications. This approach allows domain experts to directly contribute in the developing of applications without needing low level knowledge on WSN platforms and, at the same time, it allows network experts to program WSN nodes to met application requirements without specific knowledge on the application domain. Our approach also promotes the reuse of the developed software artifacts, allowing an application model to be reused across different sensor platforms and a platform model to be reused for different applications

Um modelo conceitual baseado em MDD e padrões para evolução de sistemas OA

Aspect-Oriented Software Development (AOSD) is a technique that complements the Object- Oriented Software Development (OOSD) modularizing several concepts that OOSD approaches do not modularize appropriately. However, the current state-of-the art on AOSD suffers with software evolution, mainly because aspect definition can stop to work correctly when base elements evolve. A promising approach to deal with that problem is the definition of model-based pointcuts, where pointcuts are defined based on a conceptual model. That strategy makes pointcut less prone to software evolution than model-base elements. Based on that strategy, this work defines a conceptual model at high abstraction level where we can specify software patterns and architectures that through Model Driven Development techniques they can be instantiated and composed in architecture description language that allows aspect modeling at architecture level. Our MDD approach allows propagate concepts in architecture level to another abstraction levels (design level, for example) through MDA transformation rules. Also, this work shows a plug-in implemented to Eclipse platform called AOADLwithCM. That plug-in was created to support our development process. The AOADLwithCM plug-in was used to describe a case study based on MobileMedia System. MobileMedia case study shows step-by-step how the Conceptual Model approach could minimize Pointcut Fragile Problems, due to software evolution. MobileMedia case study was used as input to analyses evolutions on software according to software metrics proposed by KHATCHADOURIAN, GREENWOOD and RASHID. Also, we analyze how evolution in base model could affect maintenance on aspectual model with and without Conceptual Model approaches

Empreendedorismo social e protagonismo juvenil: a estratégia socioeducativa do projeto Oasis de intervivência universitária

This dissertation aims to analyze the strategy of socio Intervivência University Project OASIS training of community development from the perspective of social entrepreneurship. This project was part of the context of a sort of university extension, called Intervivência University, which emerged in 2008 through the announcement MCT / CNPq / CT-Agribusiness / MDA - No. 23/2008 - Intervivência University Program. With the analysis of the OASIS project sought to identify a strategy socio, which was used elements of popular education and aimed to develop young people within the social entrepreneurship that made contributions to such has taken actions in their communities seeking range of social gains. For this, the methodological aspect, it was used in the research, a qualitative approach, case study, content analysis, semi-structured interviews and non-participant observation. In the theoretical aspect was addressed in the affairs of: social entrepreneurship, with emphasis on the concept, their differentiations of business entrepreneurship and social responsibility, and characterizing the social entrepreneur, popular education, as for the development of social entrepreneurship is the need for a new way of thinking and seeing reality for the people, and youth leadership, given the importance in the context of social action, to have young people seeking a better life for their communities. Addressed the theory were drawn categories (capital, empowerment and youth participation) and subcategories of analysis (value, attitude, behavior, trust, cooperation, civic participation) used in this work. Given the information and analysis of interviews and observations could be perceived that the strategy promoted socio Project OASIS changes, or the reaffirmation of values, resulting in new attitudes and, consequently, new behaviors in the students interviewed. Has been identified in them the development of trust and cooperation. In the aspect of civic awakening was observed in some of them only. In relation to juvenile activity was perceived to develop actions aimed at social benefits for all respondents. Thus, the OASIS Project Intervivência University made contributions to the young participants in the research aspect of social entrepreneurship, influencing them in developing activities in the social sphere in which cities are embedded

O programa de aquisição de alimentos (PAA) e as transformações socioterritoriais no Rio Grande do Norte

The Acquisition Food Program (Programa de Aquisição de Alimentos – PAA) is one of the governmental public politics proposed, in the year of 2003, that has the focus in the family-run farm in Brazil. This Program has as objectives: commercialize the familyrun farm products, bought by the government agencies; to encourage production, and promoting actions that improve quality food of people facing starving risk. This Program has been working, to some extent, as complement to the Program of family-run farm Enhancement (PRONAF). The present study has as its focus to analyse the social territorials changes observed in the state of Rio Grande do Norte, as a result of the PAA implementation. In this sense, it was promoted a discussion between territory and rules. I this work, public politics are understood as rules. This procedure was important in order to understand how the rules have influenced territorial changes, creating new social and territorial relations. In the research process was used scientific and institutional texts on the subject. It was applied a quiz with 97 PAA’s supplying farmers, and interviews were done with the Program managers in Federal (MDA), State (CONAB and EMATER) and municipal (local office of EMATER and Departments of Agriculture) levels. Social public politics were also analyzed with the aim of understand the context where PAA is situated in relation with the public politics directed to familyrun farm as a whole since 1980, within new Brazilian democratization process. With the information collected in the field, plus the secondary data, maps, charts, tables and cartograms were made. With all of this, a socio-economical profile of the family-run farm in the Rio Grande do Norte was traced, as well as the included in this research. Before this picture, we concluded that PAA indeed represents a forward in public politics directed to family-run farm. However, in spite of the importance of the Program for the family-run farmers to come of age, but in order to the rural and family-run farm reality in the Rio Grande do Norte come to present meaningful changes much still remain to be done.

ArchWiSeN: uma estratégia baseada em modelos para desenvolvimento de aplicações para redes de sensores e atuadores sem fio

Wireless Sensor and Actuator Networks (WSAN) are a key component in Ubiquitous Computing Systems and have many applications in different knowledge domains. Programming for such networks is very hard and requires developers to know the available sensor platforms specificities, increasing the learning curve for developing WSAN applications. In this work, an MDA (Model-Driven Architecture) approach for WSAN applications development called ArchWiSeN is proposed. The goal of such approach is to facilitate the development task by providing: (i) A WSAN domain-specific language, (ii) a methodology for WSAN application development; and (iii) an MDA infrastructure composed of several software artifacts (PIM, PSMs and transformations). ArchWiSeN allows the direct contribution of domain experts in the WSAN application development without the need of specialized knowledge on WSAN platforms and, at the same time, allows network experts to manage the application requirements without the need for specific knowledge of the application domain. Furthermore, this approach also aims to enable developers to express and validate functional and non-functional requirements of the application, incorporate services offered by WSAN middleware platforms and promote reuse of the developed software artifacts. In this sense, this Thesis proposes an approach that includes all WSAN development stages for current and emerging scenarios through the proposed MDA infrastructure. An evaluation of the proposal was performed by: (i) a proof of concept encompassing three different scenarios performed with the usage of the MDA infrastructure to describe the WSAN development process using the application engineering process, (ii) a controlled experiment to assess the use of the proposed approach compared to traditional method of WSAN application development, (iii) the analysis of ArchWiSeN support of middleware services to ensure that WSAN applications using such services can achieve their requirements ; and (iv) systematic analysis of ArchWiSeN in terms of desired characteristics for MDA tool when compared with other existing MDA tools for WSAN.

Efeito da Olmesartana na resposta inflamatória em modelo de mucosite intestinal em ratos

Intestinal Mucositis is inflammation and/or ulceration of mucosa of the gastrointestinal tract caused by anticancer therapies. Histologically, villous atrophy, damage to enterocytes and infiltration of inflammatory cells. Methotrexate (MTX) is a compound that depletes dihydrofolate pools and is widely used in the treatment of leukemia and other malignancies. The aim of this study was to evaluate the effect of Olmesartan (OLM), an angiotensin II receptor antagonist, on an Intestinal Mucositis Model (IMM) induced by MTX in Wistar rats. IMM was induced via intraperitoneal (i.p.) administration of MTX (7 mg/kg) for three consecutive days. The animals were pretreated with oral OLM at 0.5, 1 or 5 mg/kg or with vehicle 30 min prior to exposure to MTX, for three days. Small intestinal (duodenum, jejunum and ileum) homogenates were assayed for levels of the IL-1β, IL-10 and TNF-α cytokines, malondialdehyde and myeloperoxidase activity. Additionally, immunohistochemical analyses of MMP-2, MMP-9, COX-2, RANK/RANKL and SOCS-1 and confocal microscopy analysis of SOCS-1 expression were performed. Treatment with MTX+OLM (5 mg/kg) resulted in a reduction of mucosal inflammatory infiltration, ulcerations, vasodilatation and hemorrhagic areas (p<0.05) as well as reduced concentrations of MPO (p<0.001) and the pro-inflammatory cytokines IL-1β and TNF-α (p<0.01), and increase antiinflammatory cytosine IL-10 (p,0.05). Additionally, the combined treatment reduced expression of MMP-2, MMP-9, COX-2, RANK and RANKL (p<0.05) and increased cytoplasmic expression of SOCS-1 (p<0.05). Our findings confirm the involvement of OLM in reducing the inflammatory response through increased immunosuppressive signaling in an IMM. We also suggest that the beneficial effect of Olmesartan treatment is specifically exerted during the damage through blocking inflammatory cytosines.

Avaliação do efeito anti-inflamatório dos fármacos: atorvastatin, carvedilol, olmesartan e telmisartan, em modelo experimental da doença periodontal induzida em ratos

A periodontite é uma doença crônica inflamatória mediada por marcadores inflamatórios, tais como as citocinas: IL-1β, IL-10 e TNF-α, que provoca a destruição dos tecidos gengivais e osso alveolar, causando perda de inserção dentária e posterior perda dental. A perda óssea é causada pela ativação de prostaglandinas oriundas do ácido araquidônico, através da ação da enzima ciclooxigenase 2 (COX-2), promovendo a liberação de enzimas proteolíticas, as metaloproteinases de matriz, principalmente a MMP-2 e MMP-9, que promovem reabsorção óssea. Além disso, ocorre o desequilíbrio entre a ação de RANKL e OPG, havendo uma maior ativação de RANKL, e por consequência a maior ativação de osteoclastos e maior reabsorção óssea. Mediadores inflamatórios e espécies reativas de oxigênio (ROS) produzidos localmente possuem potencial para disseminar na corrente sanguínea e iniciar ou exacerbar doenças sistêmicas como as cardiovasculares. O tratamento atual da doença consiste em terapêutica local, mas a necessidade de estudos sobre fármacos de atuação sistêmica culminou nesta pesquisa, que realizou a avaliação dos fármacos: atorvastatin, carvedilol, olmesartan e telmisartan, quanto a sua ação anti-inflamatória sobre a doença periodontal induzida por ligadura em ratos Wistar. Os animais foram divididos em 5 grupos, para cada fármaco, separadamente: (NL) grupo não ligado, (L) grupo ligado sem tratamento, (1mg/Kg) grupo ligado que recebeu dose de 1mg/Kg de fármaco, (5 ou 6 mg/Kg) grupo ligado que recebeu dose de 5 ou 6 mg/Kg de fármaco, (10 mg/Kg) grupo ligado que recebeu dose de 10mg/Kg de fármaco. Foram realizadas avaliações: histopatológica, perda óssea alveolar, imuno-histoquímica (para COX-2, MMP-2, MMP-9, RANK-L, RANK e OPG), e ELISA (para mieloperoxidase, glutationa, malonaldeído e as citocinas: IL-1β, IL-10 e TNF-α). Os grupos tratados com olmesartana a 6 mg/Kg, e atorvastatin, carvedilol e telmisartan a 10mg/Kg, mostraram diminuição da perda óssea, redução de: MPO, MDA, IL-1β, TNF-α, MMP-2, MMP-9, COX-2, RANKL/RANK, e aumento na expressão da OPG e da IL-10.

Efeitos da terapia fotodinâmica com aluminio – cloro ftalocianina sobre a peroxidação lipídica e produtos antioxidantes em tecidos inflamados animais

Photodynamic therapy (PDT) consists of a non-toxic photosensitizing agent (FS) administration followed by a laser source resulting in a sequence of photochemical and photobiological processes that generate reactive oxygen species (ROS) that damaging cells. The present work evaluated the effects of PDT nanoemulsion-aluminum chloride phthalocyanine (AlClFc) mediated on malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels, which represent indicators involved in oxidative stress and antioxidant defenses. For this purpose, this study used 120 female rats of the Rattus norvegicus species, Wistar race, divided into 5 groups: Healthy (H), with periodontal disease (PD), with periodontal disease and treatment with FS (F), with periodontal disease and treatment with the laser (L); and periodontal disease and treatment with PDT (FL). An experimental model for represent periodontal disease (PD) was induced by ligature (split-mouth). Seven days later the induction of PD, the treatments were instituted according to the groups. In the group treated with PDT was applied 40μl FS (5μM) followed by laser irradiation diode InGaAlP (660nm, 100J / cm2). The rats were sacrificed on the 7th and 28th day after treatment and tissue specimens were removed and subjected to histological, immunohistochemical methods and enzymatic colorimetric measurements with detection by UV / VIS spectroscopy. Inflammatory changes, connective tissue disorganization and alveolar bone loss were displaying in groups with PD induced. The enzyme dosages showed that MDA levels were higher in PD induced groups, with no statistically significant differences (p> 0.05). High levels of GSH were found in groups L (p = 0.028) and FL (p = 0.028) compared with PD group, with statistically significant differences. Immunohistochemistry for SOD showed higher immunostaining in L and FL groups, compared to the PD group without statistically significant differences (p> 0.05). GPx showed lower immunoreactivity in the DP group when compared to the other groups and statistically significant differences were observed between the DPxL groups (p <0.05). TFD administered in this experiment did not induce elevation of MDA levels significantly increased the GSH levels and showed intense immunostaining pada SOD and GPx, showing that this therapy does not accentuated lipid peroxidation, however, it was able to induce effects on the antioxidant defenses processes. The LBI therapy appeared to show photomodulatory promoting effects reduction of the MDA levels, increasing GSH levels and with intense immunostaining for SOD and GPx, demonstrating that laser therapy induced antioxidant effects.

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