Isolation and partial characterization of a 30 kDa adhesin from Paracoccidioides brasiliensis

The virulence of Paracoccidioides brasiliensis can be attenuated or lost after long periods of repeated subculturing and reestablished after animal inoculation. Only one adhesin (gp43) has been described until now, among the various identified components of P. brasiliensis, and gp43 shows adhesion to laminin. Thus, the present study was designed to isolate and characterize factors putatively related to the capacity of this fungus to adhere to the host by comparing P brasiliensis samples, taken before and after animal inoculation. The two samples differed in their pattern of adhesion and invasion. The sample recently isolated from animals (Pb18b) demonstrated a greater capacity to adhere and to invade the Vero cells than the one subcultured in vitro (Pb18a). Extract from Ph18b also showed higher levels of protein expression than that from Pb18a, when two-dimensional electrophoresis gels were compared. A protein species of 30 kDa, pI 4.9, was more evident in the Pb18b extract and had properties of adhesin. Laminin, but none of the other extracellular matrix (ECM) components, such as fibronectin, collagen I and IV, bound specifically to the P. brasiliensis 30 kDa protein. The roles of 30 kDa and gp43 in cellular interactions were investigated and the adhesion of P. brasiliensis yeast cells was intensively inhibited by pre-treatment of epithelial cells with 30 kDa protein and gp43. Thus, this study presents evidence that adhesion capacity could be related to virulence, and that a 30 kDa adhesin accumulated differentially in samples with different levels of pathogenicity. This protein and its adhesion characteristics are being published for the first time and may be related to the virulence of P brasiliensis. (c) 2005 Elsevier SAS. All rights reserved.

Identification of a gene encoding adaptin-like protein in the Paracoccidioides brasiliensis genome by random amplified polymorphic DNA analysis

Paracoccidioides brasiliensis isolates are not homogeneous in their patterns of pathogenicity in animals and adhesion to epithelial cells. During this investigation, genotypic differences were observed between two samples of P. brasiliensis strain 18 yeast phase (Pbl 8) previously cultured many times, one taken before (Pb18a) and the other after (Pb18b) animal inoculation. Random amplified polymorphic DNA analysis using the primer OPJ4 distinguished Pb18b from Pbl Ba by one 308 bp DNA fragment, which after cloning and sequencing was shown to encode a polypeptide sequence homologous to the protein beta-adaptin. It is suggested, by comparison to other micro-organisms, that this protein might play an important role in the virulence of P. brasiliensis. This result demonstrates the influence of in vitro subculturing on the genotype of this organism.

Adherence and intracellular parasitism of Paracoccidioides brasiliensis in Vero cells

Paracoccidioides brasiliensis is a dimorphic fungus known to produce invasive systemic disease in humans. The 43-kDa glycoprotein of P, brasiliensis is the major diagnostic antigen of paracoccidioidomycosis and may act as a virulence factor, since it is a receptor for laminin. Very little is known about early interact-ions between this fungus and the host cells, so we developed in vitro a model system employing cultured mammalian cells (Vero cells), in order to investigate the factors and virulence mechanisms of P. brasiliensis related to the adhesion and invasion process. We found that there is a permanent interaction after 30 min of contact between the fungus and the cells. The yeasts multiply in the cells for between 5 and 24 h. Different strains of P, brasiliensis were compared, and strain 18 thigh virulence) was the most strongly adherent, followed by strain 113 (virulent), 265 (considered of low virulence) and 113M(mutant obtained by ultraviolet radiation, deficient in gp43). P. brasiliensis adhered to the epithelial cells by a narrow tube, while depressions were noticed in the cell surface, suggesting an active cavitation process. An inhibition assay was performed and it was verified that anti-gp43 serum and a pool of sera from individuals with paracoccidioidomycosis were able to inhibit the adhesion of P. brasiliensis to the Vero cells. Glycoprotein 43 (gp43) antiserum abolished 85 % of the binding activity of P. brasiliensis. This fungus can also invade the Vero cells, and intraepithelial parasitism could be an escape mechanism in paracoccidioidomycosis. (C) 2000 Editions scientifiques et medicales Elsevier SAS.

Epithelial cells treated with genistein inhibit adhesion and endocytosis of Paracoccidioides brasiliensis

Paracoccidioidomycosis is caused by Paracoccidioides brasiliensis, which although not formally considered an intracellular pathogen, can be internalized by epithelial cells in vitro and in vivo. The mechanisms used by P. brasiliensis to adhere to and invade non-professional phagocytes have not been identified. The signal-transduction networks, involving protein tyrosine kinase (PTK) and protein phosphatase activities, can modulate crucial events during fungal infections. In this study, the involvement of PTK has been investigated in P. brasiliensis adherence and invasion in mammalian epithelial cells. A significant inhibition of the fungal invasion occurred after the pre-treatment of the epithelial cells with genistein, a specific tyrosine kinase inhibitor, indicating that the tyrosine kinase pathway is involved in P. brasiliensis internalization. In contrast, when the fungus was treated, a slight (not significant) inhibition of PTK was observed, suggesting that PTK might not be the fungus' transduction signal pathway during the invasion process of epithelial cells. An intense PTK immunofluorescence labeling was observed in the periphery of the P. brasiliensis infected cells, little PTK labeling was found in both uninfected cells and yeast cells, at later infection times (8 and 24 h). Moreover, when the epithelial cells were treated with genistein and infected with P. brasiliensis, no labeling was observed, suggesting the importance of the PTK in the infectious process. These results suggest that PTK pathway participates in the transduction signal during the initial events of the adhesion and invasion processes of P. brasiliensis to mammalian epithelial cells.

Influência da fisioterapia complexa descongestiva associada à ingestão de triglicerídeos de cadeia média no tratamento do linfedema de membro superior

OBJETIVO: Verificar a influência da utilização da fisioterapia complexa descongestiva associada à dietoterapia com triglicerídeos de cadeia média (TCM) como forma de intervenção no linfedema de membro superior (MS). MÉTODOS: Para a avaliação do linfedema, foram utilizadas cirtometria, volumetria, pregas cutâneas e quantidade de água corporal total. A Escala Visual Análoga (EVA) foi utilizada para avaliar as sensações de desconforto, peso e dor no MS. Participaram deste estudo dez mulheres mastectomizadas com linfedema de MS homolateral à cirurgia, com idade média de 65,9 ± 10,4 anos e índice de massa corpórea (IMC) de 26,8 ± 3,0kg/m² que, após avaliação nutricional, foram divididas aleatoriamente em dois grupos: Grupo Controle (n= 5), submetido ao tratamento fisioterapêutico constando da terapia complexa descongestiva (massagem clássica, drenagem linfática manual, bandagem compressiva e cuidados com a pele) três vezes na semana, durante quatro semanas; Grupo TCM (n= 5), submetido ao mesmo protocolo fisioterapêutico somado ao tratamento dietético diário com ingestão de TCM, por quatro semanas. RESULTADOS: Ao final da intervenção, a análise da cirtometria e da volumetria mostraram diferenças significativas entre os grupos (< 0,05), com maior redução do linfedema no Grupo TCM. Não houve diferença significativa nos valores das pregas cutâneas e da quantidade de água corporal total. A sensação de peso no membro superior, antes e após a intervenção, foi significativamente menor (< 0,05) no Grupo TCM. CONCLUSÕES: O tratamento fisioterapêutico somado à dietoterapia com ingestão de TCM em mulheres portadoras de linfedema de MS pós-cirurgia e tratamento de câncer de mama foi efetivo na involução desta condição.

Avaliação do polimorfismo no gene da metilenotetrahidrofolato redutase e concentração de folato e vitamina B12 em pacientes portadores do HIV-1 em tratamento com anti-retrovirais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeitos do tratamento com Hidrogel na cicatrização de úlceras venosas de membros inferiores: revisão sistemática

Chronic venous disease (CVD) is evident among the chronic diseases and affects the elderly population and primarily is responsible for leg ulcers in this population. The use of dressings in the care of a venous ulcer is a fundamental part of the treatment for healing, however, evidence to assist in choosing the best dressing is scarce. The main objective of this study was to evaluate the effectiveness of treatment with hydrogel in the healing of venous ulcers using search methods, synthesis of information and statistical research through a systematic review and meta-analysis. Randomized controlled trials were selected in the following databases: CENTRAL; DARE; NHS EED; MEDLINE; EMBASE; CINAHL. Beyond these databases three websites were consulted to identify ongoing studies: ClinicalTrials.gov, OMS ICTRP e ISRCTN. The primary outcomes were analyzed: complete wound healing, incidence of wound infection and the secondary were: changes in ulcer size, time to ulcer healing, recurrence of ulcer, quality of life of participants, pain and costs of treatment. Four studies are currently included in the review with a total of 250 participants. The use of hydrogel appears to be superior to conventional dressing, gauze soaked in saline, for the healing of venous leg ulcers; 16/30 patients showed complete healing of ulcers (RR 5,33, 95%CI [1,73,16,42]). The alginate gel was shown to be more effective when compared to the hydrogel dressing in reduction of the wound area; 61,2% (± 26,2%) with alginate e 19,4% (± 24,3%) with hydrogel at the end of four weeks of treatment. Manuka honey has shown to be similar to the hydrogel dressings in percentage of area reduction. This review demonstrated that there is no evidence available about the effectiveness of the hydrogel compared to other types of dressings on the healing of venous leg ulcers of the lower limbs, thus demonstrating the need of future studies to assist health professionals in choosing the correct dressing.

O tratamento odontológico sob o olhar da criança :um estudo de representações sociais

Oral health measures directed to children are characterized by the lack of policies that consider the individual and his biopsychosocial aspects. As a consequence, it can be observed that a child´s dental experience has normally been prejudged as unpleasant and threatening. Thus, this study is aimed at investigating dental treatment from the child´s viewpoint using the theory of social representations for theoretical/methodological support. This theory was selected for being able to build practical knowledge surrounding the interrelations between social actors, the phenomenon and the world around them. The investigation was performed at the Professor José Fernandes Machado State School, located in the Ponta Negra district of Natal-RN-Brazil. The participants consisted of 30 children from the public elementary school system in the 6-10 year age group, who had undergone dental treatment within the previous year or were being treated at the time of data collection. An in-depth interview and story drawing within a theme were used as collection instruments. Thematic Content Analysis (Bardin, 2002) was used to analyze the material collected in the interviews, whereas for story drawing within a theme, in addition to content analysis, the data were also analyzed by the Coutinho model (2001). The two instruments demonstrated similarity of content as well as complementarity and contained categories associated to the model for dental treatment in children, the description of the dental office environment., the perception of the image of the dentist, as well as psychological and behavioral manifestations. The results enable us to understand determinate reactions of the children in relation to dental treatment, which has been characterized by its technical-curative approach, where the motive for seeking treatment, the manner in which it is performed and the relation of the dentist with the child are all in evidence. Thus, this study intends not only to reveal the viewpoint of children faced with dental treatment, but also to contribute to a dental practice directed towards them, in this way instilling in the professionals who attend them a more concrete awareness of the needs, anxieties and feelings of these young patients

Impacto sócio-dentário das oclusopatias e suas respectivas necessidades de tratamento na qualidade de vida de adolescentes

Nas últimas décadas, com a significativa redução da cárie dentária em crianças e adolescentes, uma maior atenção tem sido dada a outros problemas bucais como as oclusopatias. Em termos epidemiológicos no Brasil, esta tem ocupado o terceiro lugar dentre os problemas de saúde oral na população e, portanto, a inclusão destas como um problema de saúde pública se deve não apenas à sua alta prevalência, mas também ao impacto social, que interfere diretamente na qualidade de vida das pessoas. Diante do exposto é que este estudo objetiva buscar o impacto da presença e severidade das oclusopatias e suas respectivas necessidades de tratamento na qualidade de vida de adolescentes escolares de um município. Sendo assim, foram examinados 356 escolares na faixa etária de 15 -19 anos e determinadas a prevalência, severidade e necessidade de tratamento de oclusopatias através dos índices DAI, IOTN e ICON. O impacto sócio dentário das oclusopatias na qualidade de vida dos adolescentes foi analisado através do índice CS-OIDP. A fim de se verificar possíveis influências de variáveis estudadas na determinaçao do impacto na qualidade de vida das oclusopatias, estas serão coletadas através do estatus sócio-econômico, gênero, idade e tipo escola. Nos resultados parciais, relacionado a 50% da amostra, se observou uma maior prevalência do escolares de escola pública(68,1%), idade entre 15-16 anos(74,7%), gênero feminino(52,7%) e maior prevalência do SES baixo(77,5%). A análise estatística usando o teste qui-quadrado mostrou uma associação significativa do impacto da oclusopatia na qualidade de vida apenas quando avaliada com o índice ICON e o A-IOTN do examinado, comprovando uma associação da necessidade de tratamento com a presença do impacto na qualidade de vida. Com isso, busca-se comprovar e enfatizar a necessidade de se estabelecer políticas públicas que trabalhem na prevenção e interceptação dessas oclusopatias na população, melhorando, portanto, a qualidade de vida e saúde oral dos adolescentes.

Impacto sócio-dentário das ocusopatias e suas respectivas necessidades de tratamento na qualidade de vida de adolescentes

Nas últimas décadas, com a significativa redução da cárie dentária em crianças e adolescentes, uma maior atenção tem sido dada a outros problemas bucais como as oclusopatias. Em termos epidemiológicos no Brasil, esta tem ocupado o terceiro lugar dentre os problemas de saúde oral na população e, portanto, a inclusão destas como um problema de saúde pública se deve não apenas à sua alta prevalência, mas também ao impacto social, que interfere diretamente na qualidade de vida das pessoas. Diante do exposto é que este estudo objetiva buscar o impacto da presença e severidade das oclusopatias e suas respectivas necessidades de tratamento na qualidade de vida de adolescentes escolares de um município. Sendo assim, foram examinados 356 escolares na faixa etária de 15 -19 anos e determinadas a prevalência, severidade e necessidade de tratamento de oclusopatias através dos índices DAI, IOTN e ICON. O impacto sócio dentário das oclusopatias na qualidade de vida dos adolescentes foi analisado através do índice CS-OIDP. A fim de se verificar possíveis influências de variáveis estudadas na determinaçao do impacto na qualidade de vida das oclusopatias, estas serão coletadas através do estatus sócio-econômico, gênero, idade e tipo escola. Nos resultados parciais, relacionado a 50% da amostra, se observou uma maior prevalência do escolares de escola pública(68,1%), idade entre 15-16 anos(74,7%), gênero feminino(52,7%) e maior prevalência do SES baixo(77,5%). A análise estatística usando o teste qui-quadrado mostrou uma associação significativa do impacto da oclusopatia na qualidade de vida apenas quando avaliada com o índice ICON e o A-IOTN do examinado, comprovando uma associação da necessidade de tratamento com a presença do impacto na qualidade de vida. Com isso, busca-se comprovar e enfatizar a necessidade de se estabelecer políticas públicas que trabalhem na prevenção e interceptação dessas oclusopatias na população, melhorando, portanto, a qualidade de vida e saúde oral dos adolescentes.

Influência do tratamento periodontal sobre os níveis da proteína c-reativa ultrassensível em pacientes com insuficiência renal crônica em fase de prétransplante

The present study evaluated the influence of non-surgical periodontal treatment on the levels of C- reactive protein (hsCRP) in patients with chronic renal failure (CRF) in pretransplant. We conducted a controlled and randomized trial to evaluate the periodontal condition and plasma concentrations of hsCRP, albumin and transferrin in 56 dialysis patients divided into two groups: experimental and control. The study was conducted at the dental clinic of Family and Community Health s Unit (USFC), located in Onofre Lopes University Hospital (HUOL), Federal University of Rio Grande do Norte (UFRN), from December 2010 to November 2011. Severe periodontitis was the type of periodontal disease more common, affecting 78.6% of patients. Periodontal conditions, evaluated through the means of probing depth, clinical attachment level, bleeding index and plaque index, proved to be uniform for both groups at the initial examination. There were no differences in levels of inflammatory markers between the two groups. The analysis of the concentrations of hsCRP allowed classifying study participants as at high risk of developing cardiovascular disease. After completion of periodontal treatment in the experimental group, there was a statistically significant reduction of the mean of all periodontal parameters assessed; however this improvement of periodontal health was not accompanied by changes in the levels of hsCRP, albumin and transferrin in the evaluation time. Given this, the periodontal treatment did not promote the reduction of systemic inflammatory burden and risk of cardiovascular complications in patients with CRF

Dinâmica de reprodução e comportamento reprodutivo de branchoneta Dendrocephalus brasiliensis Pesata, 1921 como incremento na produção de alimento vivo para peixes ornamentais

Reproductive behavior of Dendrocephalus brasiliensis Pesta, 1921(Crustacean: Anostracan). The reproductive behavior of fresh water Anostracan has been poorly investigated in carcinology specialized literature, specifically in relation to Dendrocephlaus brasiliensis Pesta, 1921, with abundant data just about the geographical distribution of that Anostracan. The objective of this work was to know the reproductive behavior of this Anostracan, in different seasons (dry and raine). For this, ripe individuals of both sexes were collected in four pond of the of Fish farming Station of Paulo Afonso (EPPA), through monthly captures in each ponds, from December 2004 to November 2005. The type of reproduction was observed after the placing in aquariums (a) of then with males and (b) other on individually, at the nauplii phase, where they stayed for 15 days until the reproductive age. The sexual proportion was calculated through the relative frequencies of males and females, every month, for the whole collection period during 10 days. the production of cysts was related to the size of the female. The male: female ratio in the study period was 1 male:1,07 female. The proportion male:female was from 48,25% : 51,75% along the year. Concerning the type of reproduction, it was observed that this specie is characteerized by sexual reproduction

Avaliação da estimulação da medula espinhal como modelo de tratamento da doença de parkinson no primata Callithrix jacchus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Estratégia reprodutiva do peixe-voador, Hirundichthys affinis e do peixe agulha preta, Hemiramphus brasiliensis no litoral de Caiçara do Norte, Rio Grande do Norte, Brasil

The flyingfish, Hirundichthys affinis (Günther, 1866) (Exocoetidae) and ballyhoo half beak, Hemiramphus brasiliensis (Linnaeus, 1758) (Hemiramphidae) are marine fish species of commercial value, which contribute to artisanal fishery production of Rio Grande do Norte, Brazil. Besides the economic importance, these fish are important from an ecological point of view, being components in pelagic food chain, where they are the preferred prey of larger predators of high commercial value. This study aimed to determine the reproductive strategies of H. affinis and H. brasiliensis and the fishery production of H. affinis in the northern coast of Rio Grande do Norte. The results are presented in the form of six cientific articles and a review paper. The first article presents the morphometric measurements and meristic counts which confirmed the taxonomic status, population structure in length-weight and the gonad development of H. affinis through macroscopic observations of the ovaries and testes. Females of this species were bigger and heavier than males. The second article describes the reproductive tactics, such as, sex ratio, length at first sexual maturity, histological aspects of gonads, fecundity and spawning season, that indicate the reproductive strategy of H. affinis. The third article shows annual changes in the fishery production of H. affinis in Caiçara do Norte, during the period of 1993 to 2010, emphasizing the relevance of this fish in total fishery production of Rio Grande do Norte, besides registering the fall in production in the years 2008 to 2010. The fourth article describes the population structure, with respect to the length and weight, the length-weight relationship, growth type and sex ratio of Hemiramphus brasiliensis. The fifth article describes the reproductive biology of H. brasiliensis, with details about sex ratio, length at first sexual maturity, fecundity, type of breeding and spawning season. The sixth article deals with the reproductive strategy of H. brasiliensis. The review paper presents the topic of age determination of tropical fish using analysis of otoliths. Both species use the coastal waters of Caiçara do Norte for reproductive purposes. The flying fish, H. affinis migrates from the oceanic waters to the coastal waters to reproduce, coinciding with the rainy season. H. affinis presents total spawning and a seasonal reproductive strategy. However, H. brasiliensis lives and reproduces in the coastal waters displaying partial spawning and an equilibrium reproductive strategy