Novas propriedades do SKTI (Inibidor de tripsina de soja): inibição para elastase neutrofílica humana e efeitos no processo de injúria pulmonar aguda

Seeds from legumes including the Glycine max are known to be a rich source of protease inhibitors. The soybean Kunitz trypsin inhibitor (SKTI) has been well characterised and has been found to exhibit many biological activities. However its effects on inflammatory diseases have not been studied to date. In this study, SKTI was purified from a commercial soy fraction, enriched with this inhibitor, using anion exchange chromatography Resource Q column. The purified protein was able to inhibit human neutrophil elastase (HNE) and bovine trypsin. . Purified SKTI inhibited HNE with an IC50 value of 8 µg (0.3 nM). At this concentration SKTI showed neither cytotoxic nor haemolytic effects on human blood cell populations. SKTI showed no deleterious effects on organs, blood cells or the hepatic enzymes alanine amine transferase (ALT) and aspartate amino transferase (AST) in mice model of acute systemic toxicity. Human neutrophils incubated with SKTI released less HNE than control neutrophils when stimulated with PAF or fMLP (83.1% and 70% respectively). These results showed that SKTI affected both pathways of elastase release by PAF and fMLP stimuli, suggesting that SKTI is an antagonist of PAF/fMLP receptors. In an in vivo mouse model of acute lung injury, induced by LPS from E. coli, SKTI significantly suppressed the inflammatory effects caused by elastase in a dose dependent manner. Histological sections stained by hematoxylin/eosin confirmed this reduction in inflammation process. These results showed that SKTI could be used as a potential pharmacological agent for the therapy of many inflammatory diseases

Análise citogenética de células-tronco derivadas do subendotélio da veia umbilical humana

Mesenchymal stem cells (MSCs) are known as a population of multi-potential cells able to proliferate and differentiate into multiple mesodermal tissues including bone, cartilage, muscle, ligament, tendon, fat and stroma. Several applications of the study of EC can be emphasized the therapeutic techniques such as guided bone regeneration by implantation of EC in the affected site, without the need for bone grafts, using titanium as a vehicle. The process of cryopreservation is essential for the maintenance of cell cultures, since the cell line is frozen, it can be maintained in liquid nitrogen for an indefinite period and then thawed for therapeutic or experimental purposes. The aim of this study was to isolate a population of MSCs derived from the subendothelium of the umbilical vein human (MSCs-SUVH) to assess cytogenetic analysis by the possibility of appearance of chromosomal changes in two different situations: MSCs-SUVH regarding the process of cryopreservation and MSCs-SUVH grown on the surface of titanium. Flow cytometry analysis revealed that, this cell population was positive for the markers CD29, CD73 and CD90, but there was no expression of hematopoietic lineage markers, such as CD14, CD34 and CD45 and demonstrated capacity for osteogenic differentiation. The chromosomes obtained from the primary culture of MSCs-SUVH were analyzed by GTW banding technique, and results are described as guidelines to ISCN 2005. There was not the emergence of clonal chromosomal changes in the MSCs-SUVH in different situations analyzed. However one of the strings presented a balanced paracentric inversion, probably a cytogenetic constitutional alterations, which was present before and after the experimental situations that the MSCs-SUVH was submitted

Novas propriedades do SKTI (Inibidor de tripsina de soja): inibição para elastase neutrofílica humana e efeitos no processo de injúria pulmonar aguda

Seeds from legumes including the Glycine max are known to be a rich source of protease inhibitors. The soybean Kunitz trypsin inhibitor (SKTI) has been well characterised and has been found to exhibit many biological activities. However its effects on inflammatory diseases have not been studied to date. In this study, SKTI was purified from a commercial soy fraction, enriched with this inhibitor, using anion exchange chromatography Resource Q column. The purified protein was able to inhibit human neutrophil elastase (HNE) and bovine trypsin. . Purified SKTI inhibited HNE with an IC50 value of 8 µg (0.3 nM). At this concentration SKTI showed neither cytotoxic nor haemolytic effects on human blood cell populations. SKTI showed no deleterious effects on organs, blood cells or the hepatic enzymes alanine amine transferase (ALT) and aspartate amino transferase (AST) in mice model of acute systemic toxicity. Human neutrophils incubated with SKTI released less HNE than control neutrophils when stimulated with PAF or fMLP (83.1% and 70% respectively). These results showed that SKTI affected both pathways of elastase release by PAF and fMLP stimuli, suggesting that SKTI is an antagonist of PAF/fMLP receptors. In an in vivo mouse model of acute lung injury, induced by LPS from E. coli, SKTI significantly suppressed the inflammatory effects caused by elastase in a dose dependent manner. Histological sections stained by hematoxylin/eosin confirmed this reduction in inflammation process. These results showed that SKTI could be used as a potential pharmacological agent for the therapy of many inflammatory diseases

Purificação e caracterização parcial de uma Fucana C de Dictyota mentrualis e estudo do seu efeito antiinflamatório e nociceptivo

In recent years, sulfated polysaccharides from marine algae have emerged as an important class of natural biopolymers with potential application in human and veterinary health care, while taking advantage of the absence of potential risk of contamination by animal viruses. Among these, fucans isolated from the cell walls of marine brown alga have been study due to their anticoagulant, antithrombotic, anti-inflammatory and antiviral activities. These biological effects of fucans have been found to depend on the degree of sulfation and molecular size of the polysaccharide chains. In the present study, we examined structural features of a fucan extracted from brown alga Dictyota menstrualis and its effect on the leukocyte migration to the peritoneum. The sulfated polysaccharides were extracted from the brown seaweed by proteolytic digestion, followed by sequential acetone precipitation producing 5 fractions. Gel lectrophoresis using 0.05 M 1,3-diaminopropane-acetate buffer, pH 9.0, stained with 0.1% toluidine blue, showed the presence of sulfated polysaccharides in all fractions. The chemical analyses demonstrated that all fractions are composed mainly of fucose, xylose, galactose, uronic acid, and sulfate. Electrophoresis in agarose gel in three different buffers demonstrated that the fraction 2.0v have only one population of fucan. This compound was purify by exclusion molecular. It has shown composition of fucose, xilose, sulfate and uronic acid in molar ration of 1.0: 1.7: 1.1: 0.5 respectively. The effect of this heterofucan on the leukocyte migration was observed 6h after zymozan (mg/g) administration into the peritoneum. The heterofucan showed higher antimigratory activity, it decrease the migration of leukocyte in 83.77% to peritoneum. The results suggest that this fucan is a new antimigratory compound with potential pharmacological appications

Avaliação das atividades anti-inflamatória, anticoagulante e antiproliferativa do inibidor de quimotripsina das sementes de erythrina velutina (EvCI)

Studies indicate that several components were isolated from medicinal plants, which have antibacterial, antifungal, antitumor and anti-inflammatory properties. Sepsis is characterized by a systemic inflammation which leads to the production of inflammatory mediators exacerbated by excessive activation of inflammatory cells and disseminated intravascular coagulation (DIC), in which the human neutrophil elastase plays an important role in its pathogenesis. Several epidemiological studies suggest that components of plants, especially legumes, can play a beneficial role in reducing the incidence of different cancers. A chymotrypsin inhibitor of Kunitz (Varela, 2010) was purified from seeds of Erythrina velutina (Mulungu) by fractionation with ammonium sulfate, affinity chromatography on Trypsin-Sepharose, Chymotrypsin-Sepharose and ion exchange chromatography on Resource Q 1 ml (GE Healthcare) in system FPLC / AKTA. The inhibitor, called EvCI, had a molecular mass of 17 kDa determined by SDS-PAGE. The purified protein was able to inhibit human neutrophil elastase (HNE), with an IC50 of 3.12 nM. The EvCI was able to inhibit both pathways of HNE release stimulated by PAF and fMLP (75.6% and 65% respectively). The inhibitor also inhibited leukocyte migration in septic mice about 87% and prolonged the time of coagulation and inhibition factor Xa. EvCI showed neither hemolytic activity nor cytotoxicity. EvCI showed a selective antiproliferative effect to HepG2 cell lines with IC50 of 0.5 micrograms per milliliter. These results suggest EvCI as a molecule antagonist of PAF / fMLP and a potential use in fighting inflammation related disorders, disseminated intravascular coagulation (DIC) and cancer

Estudo clínico-laboratorial e histopatológico de cães naturalmente infectados por Leishmania chagasi com diferentes graus de manifestação física

Canine Visceral Leishmania (CVL) is an important zoonotic disease that has a world wide distribution and has a large impact on public health on the American Continent, especially in Brazil, where the nature of endemic diseases in humans affects a large part of the nation. The influence of the prevalence of CVL in the increased rate of human cases in endemic areas and in the unleashing of epidemic outbreaks shows the need for a more profound understanding, that would generate significant advances in the current measures used to control the reservoirs of sickness that are practiced by the Programa Nacional de Vigilância e Controle da Leishmaniose Visceral. The present work describes and compares the clinical-laboratorial and histopathological findings of twenty-three dogs that were naturally infected by Leishmania chagasi, from endemic areas in metropolitan Natal, Rio Grande do Norte, Brazil. These animals, that were selected and given physical and serological exams (IFI and ELISA rK-39), were classified according to the degree of clinical severity and had blood samples drawn (whole blood and serum) for a complete hemogram and a coagulogram to be done as well as biochemical tests for kidney and liver function. The confirmation of infection by L. chagasi was done after the euthanasia of the animals, through the direct demonstration of the parasite in the impression of the spleen and liver crowned with GIEMSA and through a cultivation by means of NNN/Schneider. According to the clinical evaluation, the animals were classified as asymptomatic (7), oligosymptomatic (7) and polysymptomatic (9). Among the animals that were chosen to be autopsied, there were 2 asymptomatic, 3 oligosymptomatic and 3 polysymptomatic, for the purpose of studying their histopathology, having collected fragments of the spleen, liver, kidneys and skin and were fixed in 10% tamponed formol. The comparison between the average parameters of the clinical-laboratory tested animals in the groups was done through the Student t test (a<0.05). The main clinical signals observed were lymphadenomegaly, alopecy, dermatitis, exfoliation, cutaneous ulcers, onicogriphosis and emaciation. The main clinical-laboratorial alterations established, mainly in the polysymptomatic group, were anemia, hyperproteinemia, hyperglobulinemia, alterations in the albumin/globulin ratio and increased ALT activity. Renal alterations were not verified (urea and creatinine levels were normal). Thrombocytopenia was observed in three clinical groups. However, the other indicators of coagulation function (TAP and TTPA) did not have abnormal variations. There were inflammatory infiltrations and leishmania amastigotes in the skin of polysymptomatic dogs, however, they were not found in the skin of asymptomatic animals. Hypertrophy and hyperplasia of the phagocyte mononuclear system, leishmania amastigote parasites were found in the macrophages, extramedullary hematopoiesis and degenerative alterations were detected in the spleen and liver of 8 of the animals submitted to histopathological exams. In accord with these results, it was demonstrated that the expected alterations in the hematological and biochemical parameters in function of their viscerotropic nature of CVL are mainly observed in the more advanced stages of the disease. The absence of inflammatory infiltration and parasite load in the skin suggest that infected animals without symptoms may have an importance irrelevant to the infectiousness of the vector

Simulador radiográfico antropomórfico de mama humana

Mammography is a diagnostic imaging method in which interpretation depends on knowledge of radiological aspects as well as the clinical exam and pathophysiology of breast diseases. In this work a mammography phantom was developed to be used for training in the operation of mammographic x-ray equipment, image quality evaluation, self-examination and clinical examination of palpation. Polyurethane was used for the production of the phantoms for its physical and chemical properties and because it is one of the components normally used in prostheses. According to the range of flexibility of the polyurethane, it was possible to simulate breasts with higher or lower amount of adipose tissue. Pathologies such as areolar necrosis and tissue rejection due to surgery reconstruction after partial mastectomy were also simulated. Calcifications and nodules were simulated using the following materials: polyethylene, poly (methyl methacrylate), polyamide, polyurethane and poly (dimethyl silicone). Among these, polyethylene was able to simulate characteristics of calcification as well as breast nodules. The results from mammographic techniques used in this paper for the evaluation of the phantoms are in agreement with data found in the literature. The image analyses of four phantoms indicated significant similarities with the human skin texture and the female breast parenchyma. It was possible to detect in the radiographic images produced regions of high and low radiographic optical density, which are characteristic of breasts with regions of different amount of adipose tissue. The stiffnesses of breast phantoms were adjusted according to the formulation of the polyurethane which enabled the production of phantoms with distinct radiographic features and texture similar to human female breast parenchyma. Clinical palpation exam of the phantoms developed in this work indicated characteristics similar to human breast in skin texture, areolar region and parenchyma

Determinantes envolvidos na resposta imune celular humana à infecção por Leishmania infantum chagasi

Visceral leishmaniasis (VL) is a disease caused by protozoa of the Leishmania donovani complex, whose infection has clinical spectrum ranging from asymptomatic infection to active disease characterized by fever, cachexia, hepatosplenomegaly, and immunosuppression. The healing or protective immunity require an antigen-specific type 1. The Montenegro skin test (DTH) has been interpreted as a marker of protective immunity. However, there is no known correlation between the DTH response to type 1 and DTH and immunity of type 1 are maintained in the long term. Thus, a longitudinal study of 8 years, nested in a cohort family held in Brazil, documented the status of DTH and cytokine production by peripheral blood mononuclear cells in response to antigen-specific stimulation. This study was the interdisciplinary approach of physicians, biochemists, nutritionists, veterinary medicine, biology and statistics. The results show that 46.2% of subjects were analyzed DTH positive at baseline. The prevalence of positive and DTH induration size increased with age (p = 0.0021). 15.7% of individuals positive DTH "retro-converted" the negative and 50.4% (64) of individuals negative DTH became positive. The size of DTH induration was correlated significantly with the antigen-induced production of IFN-γ (r = 0.6186, p = 0.0001). IL-6 was secreted at higher levels in peripheral blood mononuclear cells of individuals who "retro-converted" DTH positive to negative than individuals who remained stable DTH status (p = 0.005). Thus, IFN-γ produced by peripheral blood mononuclear cells, may be a surrogate marker for protective immunity instead of the DTH response. In addition, differences in innate immune response may determine whether individuals maintain or eliminate the infection by Leishmania infantum chagasi in asymptomatic patients

A (bio)ética e a Odontologia: os (des)caminhos de uma formação humana

The present research carried out from three national dentistry magazines published in the period between 1990 and 2004, has as a goal to analyze how bioethics has been approached in this area, not only identifying the main concerns and tendencies, but also aiming to learn how this knowledge is produced and divulged in the dentistry circuit. We have articulated a quantitative-qualitative approach, studying 2995 articles. The articles were selected and assorted in twenty thematic categories, through their titles and key words. The analysis of the empiric material shows that, although there is a growth tendency of this discussion, little has been published about this theme (1,9%). Besides, it seems that there is an arrhythmia between the present bioethic approach in dentistry and the present life demands, where the deontology and legalist focuses are predominant, seeming to correspond to the inner aspects of the profession alone. In spite of this, through the qualitative approach it was possible to identify ways to build a more complex and integral odontological formation and practice. Within the conclusions, we still point out, that, this investigation, even face to its limitations, seems to offer subsidies for reflection and further studies about the theme, working as a parameter to keep up with the evolution of the bioethic thinking in the Odontology

A experiência humana como estratégia : a comunicação como jogo & o jogo como comunicação

En la interface de las Ciencias Sociales y de la Comunicación, la tesis expone una discusión sobre la Comunicación Social como área de conocimiento y discute las bases fenomenológicas de la producción de significado utilitario sobre el mundo. Para dar cuenta de esa reflexión se tiene como referencia la película Slumdog Millionaire? del director Danny Boyle. La experiencia vivida por el personaje principal llamado Jamal Malik sirve de apoyo a su éxito en un programa de TV cuyos desafíos consigue vencer uno a uno y transformarse en un millonario. El film, y en consecuencia la trayectoria de Jamal, es el operador cognitivo para que se presente la estrategia del personaje que transforma su conocimiento experimental en conocimiento pertinente, objetivo. La investigación sirve de base para ofrecer el argumento central de que solo existe conocimiento por medio de la experiencia vivida. A partir de esa concepción, la comunicación es vista en este ensayo como entrecruzamiento de caminos y nudos que se asemejan a un rizoma donde cada construcción de sentido, cada palabra que gana significación representa diversos enlaces de esos nudos sistémicos. Aseguramos que la comunicación es una condición sine qua non a los humanos y puede ser comprendida por la paradoja natural-artificial. Es de esta paradoja, que presentamos una contribución de la complejidad de la comunicación creyendo de forma dialógica que sí, hay una condición innata de la comunicación (y así mismo post humana) concomitantemente con la hipótesis de la artificialidad comunicacional. Corroboramos así, la idea de que el conocimiento fenomenológico de la comunicación es un juego de la construcción científica y, por lo tanto, un juego de la humanidad. En este mismo juego podemos comprender la comunicación por intermedio de algunos macro conceptos de la compleja actividad de comunicar (retroalimentación, recursividad y holograma) constelando así una estrategia de ligazón/estímulo/respuesta que nos permite conocer bien y saber organizar mejor el conocimiento adquirido en la práctica. La tesis tiene como interlocutores principales Claude Lévi-Strauss, Edgar Morin, Jean-Marie Pelt, Norval Baitello Junior y Vilém Flusser, entre otros

Sobre a dignidade humana: prelúdio para uma abertura das ciências jurídicas

The thesis aims at discussing and analyzing the Principle of Human Dignity in its multidimensionality, in a way not exclusively individual and anthropocentrical, but rather intersubjective and planetary, which implies to say that such understanding transcends the human itself, in order to contemplate the dignity of an individual connected to the world. In this context, the thesis proposes stretching the disciplinarity of the Law, unfolding the Legal Science to a more thorough perception of the human condition, encompassing the individual, social, anthropolitical and anthropoetical dimensions. It is not about, however, resorting to an abstract idea, but rather undertaking the construction of the concrete universality of such understanding, which translates into contextualizing the face of a planetary identity of the man, also taking into account its triunic nature which encompasses the dialogical and complementary relations amongst individual, society and species

Brincar, atividade humana e ferramenta pedagógica no contexto escolar da região metropolitana de natal

This thesis talks about the use of guided educational tools in play and playful in public schools in the Metropolitan Region of Natal (MRN), contributing to the teaching and students learning of basic education in the contents of portuguese and mathematics. We assume that the play is an activity / human need and therefore implies the proper development of children in physical, psychological, biological, cultural, social and historical aspects. We consider the mediation exercised by the teacher in the classroom, and the affection existing between teacher and student, sine qua non terms, so that the truth and fact of the process of teaching and learning occur. Thus, theoretically collaborated Paulo Freire, with his dialogical vision; Lev S. Vygotsky says that we learn and why we develop; Lev S. Vygotsky that states that we learn and for that we develop ourselves; Pierre Bourdieu and the concept of habitus, as something embedded and therefore procedural, and the cultural capital that needs to be fed and (re)meaning at school; Luiz Pereira, Bernard Lahire, Gilles Brougère and others brought their theoretical contributions. The empirical field of research was composed by the Municipal School Professor Ulysses de Góis, located in the neighborhood of Nova Descoberta in Natal, and the Municipal School José Horacio de Góis, located in the community of Guanduba, in São Gonçalo do Amarante, municipalities of the Metropolitan Region of Natal (MRN). We used as methodology the collaborative-action-research as a possible of effective participation of the research subjects, imputing them a voice and performance in the process, and not considering them just observers. The results indicate the effectiveness of the pedagogical tools in play and playful to the learning of students, but that alone is not able to solve all the problems of the school, other referrals need to be secured, as the planning of the actions to be developed in the school and in the classroom, systematized pedagogical orienttion for faculty, family participation/involvement in the school life of students, among other actions that need to be weighted so that education fulfills its role and promote the emancipation of the subject, because in the freireana liberating perspective, "the reading of the world preced the lecture of the word"

Proteção da relação de emprego sob o prisma da dignidade da pessoa humana

This work aims to show that the protection of the employment relation is one of the determining factors to respect the principle of the human dignity. The goal is initially to show the devaluation of work from antiquity to the early twentieth century, when the constitutions began to standardize protective devices. This way, the consecration of the social labor rights in the Constitution of 1988 represents the culmination of the historical achievements. This work demonstrates that such rights can not be reduced or suppressed by political conveniences, once these rights are included in the list of immutable clauses. It is displayed that to achieve the fundamental right to work is not well advised to encourage the creation of jobs that maculates the worker s dignity. The outsourced work is, therefore, a classic example of the advancement of precarious forms of contemporary labor. It is inferred that the presence of various forms of harassment results in a degradation of working environment, bringing about dire consequences on professional and personal life of the worker. Thus, decent work must be the appropriate benchmark for the creation of new jobs. It is also shown that the flexibilization of the propaganda rights by certain pressure groups has as main goal to reduce or eliminate rights, based on fallacious data depicting an increase of competitiveness and jobs. In addition, the flexibility implies a growth of the precarization of the work - a reality felt by many workers subjected to such a situation due to the unemployment phenomenon whose origin is not in the protectionism of the norms. It is necessary to expand and structure the constitutionally legitimate institutions to monitor and curb the precarized work, as well as all practices that go against the dignity of the worker. It is also shown the loss of power of the sindicates in the last few years as a consequence of the pulverization workes and the capital attacks through the productive restructure whose outsourcings and privatizations are notorious examples

A materialização da dignidade da pessoa humana e o cumprimento das penas privativas de liberdade

The current research come from need to analyze possibilities to materialize human dignity principle during freedom curtail penalties fullfilment, abreast finding that internal and international regulations dictate this is the way to be tread by Brazilian penitentiary system, however, verily, indignity, assistance missing, overcrownding, crime, in the end, barbarie reigns. The work will analyze two strands in order to effective the mentioned principle: the state responsability optics, such in internal scope, as internationally, abreast historical omission in satisfy Constitution, international treaties and laws; and also indicating penal execution alternative methods adoption as a way, bringing to fore a case study - called "apaqueano" method. With such desideratum will bring, first of all, considerations about consolidation process of human dignity principle, its concept and essential content. Furthermore, will address historical and philosophical evolution of freedom curtail penalties. As it follows, will be done an approach about constitutional and underconstitutional legislation that disciplines penalties fullfiling in Brazil, analyzing their main aspects, emphasizing the possibility to charge Brazilian state for disregarding mentioned standards. Next, will also be started a critical analyzis about international regulations, which forbids diminishing or cruel penalties or treatments, approaching human rights international treaties and conventions ratified by Braziland their incorporation and effectiveness in local Law, emphasizing monitoring forms and country international charging possibility for disregarding international regulations. Lastly, will advance to the real possibility to materialize human dignity principle in penalties fullfiling, based in a case study verification - the APAC (Associação de Proteção e Assistência aos Condenados) called method, analyzing the various theories about penalties grounding, with emphasis in their ressocializing function, as well as traditional penitentiary systems, and the theory adopted by vernacular order, in desideratum to contribute to improve national penitentiary system chaotic situation

Dignidade da pessoa humana e medidas sócio-educativas do Estatuto da Criança e do Adolescente

The main goal of this work is to verify the presence of the principle of human dignity, determined by the Constitution of 1988, when Socio Educational Acts from the Statute of Children and adolescent were put in practice, focusing on the treatment designated to the youth whose acts were against the law in the city of Natal, as well as the difficulties to match the legal texts and its reality. It is common knowledge that the law for children and adolescent was based in the principle of human dignity, for its institutes say so. But would the Law match the practice of Socio Educational Acts? Or this law would be an example of good intentions that never left the paper and became reality? First there is an approach on the human dignity principle, with its definition and limitation, according to a theory about the theme. Afterwards it is made a connection between human rights and the principle of human dignity considering historical and social features, for the law is also a reflect of these transformations, we try to show the different laws the country had until today, concerning the children inflicted by poverty and those whose acts were against the law, since the rodas dos expostos, the phase of irregular situation, in which the children were arrested simply because they were beggars, until present time. The theory aspects are shown beside a field research made with the adolescent and staff from CEDUC/Natal, producing a critical view about the subject and showing some solutions for the problems found. At last, it is made a critical analyses of the problems detected on the field research, and, in some cases, a suggestion is given to change the reality