Purification and characterization of the biological effects of phospholipase A(2) from sea anemone Bunodosoma caissarum

Sea anemones contain a variety of biologically active substances. Bunodosoma caissarum is a sea anemone from the Cnidaria phylum, found only in Brazilian coastal waters. The aim of the present work was to study the biological effects of PLA(2) isolated from the sea anemone B. caissarum on the isolated perfused kidney, the arteriolar mesenteric bed and on insulin secretion. Specimens of B. caissarum were collected from the Sao Vicente Channel on the southern coast of the State of São Paulo, Brazil. Reverse phase HPLC analysis of the crude extract of B. caissarum detected three PLA(2) proteins (named BcPLA(2)1, BCPLA(2)2 and BcPLA(2)3) found to be active in B. caissarum extracts. MALDI-TOF mass spectrometry of BcPLA(2)1 showed one main peak at 14.7 kDa. The N-terminal amino acid sequence of BcPLA(2)1 showed high amino acid sequence identity with PLA(2) group III protein isolated from the Mexican lizard (PA23 HELSU, HELSU, PA22 HELSU) and with the honey bee Apis mellifera (PLA(2) and 1POC_A). In addition, BcPLA(2)1 also showed significant overall homology to bee PLA(2). The enzymatic activity induced by native BCPLA(2)1 (20 mu g/well) was reduced by chemical treatment with p-bromophenacyl bromide (p-BPB) and with morin. BcPLA(2)1 strongly induced insulin secretion in presence of high glucose concentration. In isolated kidney, the PLA(2) from B. caissarum increased the perfusion pressure, renal vascular resistance, urinary flow, glomerular filtration rate, and sodium, potassium and chloride levels of excretion. BcPLA(2)1, however, did not increase the perfusion pressure on the mesenteric vascular bed. In conclusion, PLA(2), a group III phospholipase isolated from the sea anemone B. caissarum, exerted effects on renal function and induced insulin secretion in conditions of high glucose concentration. (C) 2009 Elsevier Ltd. All rights reserved.

Modifiable risk factors for overweight and obesity in children and adolescents from São Paulo, Brazil

Background: Brazil is currently experiencing a nutrition transition: the displacement of traditional diets with foods high in saturated fat, sodium, and cholesterol and an increase in sedentary lifestyles. Despite these trends, our understanding of child obesity in Brazil is limited. Thus, the aims of this study were (1) to investigate the current prevalence of overweight and obesity in a large sample of children and adolescents living in São Paulo, Brazil, and (2) to identify the lifestyle behaviors associated with an increased risk of obesity in young Brazilians.Methods: A total of 3,397 children and adolescents (1,596 male) aged 7-18 years were randomly selected from 22 schools in São Paulo, Brazil. Participants were classified as normal weight, overweight, or obese based on international age-and sex-specific body mass index thresholds. Selected sociodemographic, physical activity, and nutrition behaviors were assessed via questionnaire.Results: Overall, 19.4% of boys and 16.1% of girls were overweight while 8.9% and 4.3% were obese. Two-way analysis of variance revealed that the prevalence of overweight and obesity was significantly higher in boys and in younger children when compared to girls and older children, respectively (P < 0.05 for both). Logistic regression analysis revealed that overweight was associated with more computer usage, parental encouragement to be active, and light soft drink consumption after controlling for differences in sex, age, and parental education (P < 0.05 for all). Conversely, overweight was associated with less active transport to school, eating before sleep, and consumption of breakfast, full-sugar soft drinks, fried food and confectionery (P < 0.05 for all).Conclusions: Our results show that obesity in São Paulo children and adolescents has reached a level equivalent to that seen in many developed countries. We have also identified three key modifiable factors related to obesity that may be appropriate targets for future intervention in Brazilian youth: transport mode to school, computer usage, and breakfast consumption.

An immunocytochemical and in situ hybridization analysis of annexin 1 expression in rat mast cells: Modulation by inflammation and dexamethasone

The presence and localization of the anti-inflammatory protein annexin 1 (also known as lipocortin 1) in perivenular rat mast cells was investigated here. Using the rat mesenteric microvascular bed and a combination of morphologic techniques ranging from immunofluorescence to electron microscopy analyses, we detected the presence of annexin 1 in discrete intracellular sites, both in the nucleus and in the cytoplasm. In resting mast cells, most of the protein pool (approximately 80% of the cytosolic portion) was localized to cytoplasmic granules. In agreement with other cell types, treatment of rats with dexamethasone (0.2 mg/kg, ip) increased annexin 1 expression in mast cells, inducing a remarkable appearance of dusters of protein immunoreactivity. This effect was most likely the result of de novo protein synthesis as determined by an increase in mRNA seen by in situ hybridization. Triggering an ongoing experimental inflammatory response (0.3 mg of carrageenin, ip) increased annexin 1 mRNA and protein levels. In conclusion, we report for the first time the localization of annexin 1 in connective tissue mast cells, and its susceptibility not only to glucocorticoid hormone treatment, but also to an experimental acute inflammatory response.

Mixing and motion of rice particles in a rotating drum

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of Osteopenia in Autogenous Bone Graft Healing With or Without Expanded Polytetrafluoroethylene Membranes: Histologic and Histomorphometric Study in Rats

Purpose: The aim of this study was to quantitatively evaluate and qualitatively describe autogenous bone graft healing with or without an expanded polytetrafluoroethylene (e-PTFE) membrane in ovariectornized rats. Materials and Methods: Eighty Wistar rats, weighing approximately 300 g each, were used. A graft was obtained from the parietal bone and fixed to the sidewall of each animal's left mandibular ramus. The animals were randomly divided into four experimental groups (n = 20 in each group): group 1, sham operated and autogenous bone graft only- group 2, sham operated and autogenous bone graft covered by e-PTFE membrane; group 3, ovariectornized (OVX) and autogenous bone graft only- group 4, OVX and autogenous bone graft covered by e-PTFE membrane. The animals were sacrificed at five different time points: immediately after grafting or at 7, 21, 45, or 60 days after grafting. Histologic examination and morphometric measurement of the sections were performed, and values were submitted to statistical analyses. Results: Both groups (sham and OVX) experienced loss of the original graft volume when it was not covered by the membrane, whereas use of the membrane resulted in additional bone formation beyond the edges of the graft and under the membrane. Histologic analysis showed integration of the grafts in all animals, although a larger number of marrow spaces was found in OVX groups. Conclusions: Association of bone graft with an e-PTFE membrane resulted in maintenance of its original volume as well as formation of new bone that filled the space under the membrane. Osteopenia did not influence bone graft repair, regardless of whether or not it was associated with e-PTFE membrane, but descriptive histologic analysis showed larger numbers of marrow spaces in the bone graft and receptor bed and formation of new bone in the OVX animals. INT J ORAL MAXILLOFAC IMPLANTS 2009;24:1074-1082

Tomographic, Histological, and Immunohistochemical Evidences on the Use of N-Butyl-2-Cyanoacrilate for Onlay Graft Fixation in Rabbits

Background: The bone tissue responses to Cyanoacrylate have been described in the literature, but none used N-butyl-2-cyanoacrilate (NB-Cn) for bone graft fixation. Purpose: The aims of the study were: (a) to analyze the bone grafts volume maintenance fixed either with NB-Cn or titanium screw; (b) to assess the incorporation of onlay grafts on perforated recipient bed; and (c) the differences of expression level of tartrate-resistant acid phosphatase (TRAP) protein involved in bone resorption. Materials and Methods: Eighteen New Zealand White rabbits were submitted to calvaria onlay grafting on both sides of the mandible. On one side, the graft was fixed with NB-Cn, while on the other hand the bone graft was secured with an osteosynthesis screw. The computed tomography (CT) was performed just after surgery and at animals sacrifice, after 1 (n=9) and 6weeks (n=9), in order to estimate the bone grafts volume along the experiments. Histological sections of the grafted areas were prepared to evaluate the healing of bone grafts and to assess the expression of TRAP protein. Results: The CT scan showed better volume maintenance of bone grafts fixed with NB-Cn (p≤0.05) compared with those fixed with screws, in both experimental times (analysis of variance). The immunohistochemical evaluation showed that the TRAP expression in a 6-week period was significantly higher compared with the 1-week period, without showing significant difference between the groups (Wilcoxon and Mann-Whitney). Histological analysis revealed that the NB-Cn caused periosteum damage, but provided bone graft stabilization and incorporation similar to the control group. Conclusion: The perforation provided by screw insertion into the graft during fixation may have triggered early revascularization and remodeling to render increased volume loss compared with the experimental group. These results indicate that the NB-Cn possesses equivalent properties to titanium screw to be used as bone fixation material in osteosynthesis. © 2010 Wiley Periodicals, Inc.

Healing Process of Autogenous Bone Graft in Spontaneously Hypertensive Rats Treated With Losartan: An Immunohistochemical and Histomorphometric Study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A new approach to the granulation of beta-cyclodextrin inclusion complexes

Cyclodextrins (CDs) are annular oligosaccharides containing 6-12 glucose unities joined together by alpha-1,4 bonds. They have a conical-truncated shape with a lipophilic cavity in which different molecules can be included resulting in a stable inclusion complex. The cyclodextrins have been widely applied in pharmaceutical technology with the objective of increasing the solubility, stability and bioavailability of drugs in different pharmaceutical dosage forms, such as tablets. In order to obtain beta-CD tablets, liquid dispersions of drug/beta-CD are usually submitted to different drying processes, like spray-drying, freeze-drying or slow evaporation, being this dry material added to a number of excipients. However, such drying processes can generate particulate materials showing problems of flow and compressibility, needing their conversion into granulates by means of wetting with granulation liquid followed by additional drying. In this work, the main objective was to evaluate the preparation of tablets without the need of this additional drying step. For this purpose an aqueous dispersion containing acetaminophen/beta-CD complex and cornstarch was dried using a spouted bed and the obtained granules were compressed in tablets. Acetaminophen was used as model drug due to its low water solubility and the inexpensive and widely available cornstarch was chosen as excipient. Acetaminophen powder was added into a beta-cyclodextrin solution prepared in distilled water at 70 degrees C. Stirring was kept until this dispersion cooled to room temperature. Then cornstarch was added and the resulting dispersion was dried in spouted bed equipment. This material was compressed into tablets using an Erweka Korsh EKO tablet machine. This innovative approach allowed the tablets preparation process to be carried out with fewer steps and represents a technological reliable strategy to produce beta-cyclodextrin inclusion complexes tablets. (C) 2010 Elsevier By. All rights reserved.

Comunicação em saúde: algumas reflexões a partir da percepção de pacientes acamados em uma enfermaria

Investigam-se imagens e significados sobre experiências vivenciadas por pacientes no processo de atendimento/tratamento em uma enfermaria, visando problematizar questões éticas no cotidiano das práticas de saúde, que possam contribuir para refletir, no espaço da formação profissional, sobre situações que, muitas vezes, se naturalizam pela urgência da rotina. O trabalho consistiu em entregar câmaras fotográficas a cinco pacientes acamados, internados em uma Enfermaria de Ortopedia de um hospital universitário, no segundo semestre do ano de 2001, para que registrassem, pela fotografia, experiências que vivenciavam na condição de pacientes, procurando responder às perguntas: o que significa estar na cama sem poder sair dela? O que é bom e o que é ruim em sua rotina na Enfermaria? Buscando compreender o material pela ótica dos pacientes, apresenta-se aqui parte da experiência registrada e relatada. Os resultados obtidos apontam que entre as maiores dificuldades enfrentadas pelos pacientes entrevistados estão a dependência, gerando constrangimento e desconforto, e a falta de informação, gerando insegurança e ampliando a dependência.

Treatment of venous ulcers with fibrin sealant derived from snake venom

Venous ulcers of the lower limbs complicated by infection or chronicity represent a serious public health problem. The elevated number of those afflicted burdens the health services, interferes in quality of life and causes absenteeism. Although there are 2,500 items on the market, ranging from the simplest dressing up to the most complex types of dressing, treatment remains a challenge. Among the substances used, fibrin sealant is the one that promotes diminution of bacterial colonization and of edema, controls hemorrhaging, alters the pain threshold by protecting the nerve endings, hydrates the wound bed and forms granulation tissue that favors healing. Its disadvantages include higher cost and utilization of human fibrinogen that can transmit infectious diseases. The Center for the Study of Venoms and Venomous Animals (CEVAP) at São Paulo State University (UNESP) developed a new sealant made up of fibrinogen extracted from large animals and from an enzyme obtained from snake venom. The present study, developed in the Health Education Clinic (CEPS) of Sacred Heart University (USC) aimed to evaluate the effect of the new sealant on the healing process of venous ulcers in 24 adult patients, seven of whom were male and 17 female. Two study groups were formed as follows: Group 1 (G1) - control group of 11 patients treated with essential fatty acid (EFA) and Unna's boot, and Group 2 (G2) - 13 patients treated with essential fatty acid (EFA), fibrin sealant and Unna's boot. The follow-up lasted eight weeks and the sealant was applied at only the first and fourth weeks. The results showed that Group 2 presented worse lesion conditions as to healing, but, when comparing the two groups, it was noteworthy that the the sealant was effective in healing venous ulcers. There is evidence that the new sealant is recommended for leg ulcers with the following advantages: ease of application, preparation of the wound bed, diminution of pain and a higher number of discharges in the eighth week. More important, other positive characteristics are non-transmission of infectious diseases, absence of adverse reactions, and economic advantage of being produced by Brazilian technology. Finally, it is suggested that the weekly application of sealant, for at least eight weeks, could improve the healing process and consequently life quality.

Segurança e efetividade no tratamento do pterígio usando infiltração de 5-fluoruracila no intraoperatório

OBJETIVO: Avaliar a segurança e a efetividade do uso do 5-fluoruracila (5-FU) como tratamento adjuvante do pterígio, aplicado sob a forma de infiltração subconjuntival, no período intraoperatório. MÉTODOS: Foram avaliados prospectivamente 125 indivíduos (125 olhos) portadores de pterígio. Os indivíduos foram operados segundo a técnica de retalho de deslizamento e receberam, ao final do procedimento, injeção subconjuntival de 0,2 mL de 5-FU (25 mg/mL). Foram anotados os dados do paciente como idade, sexo, profissão, características da lesão (primário ou recidivado, tamanho, carnoso ou involutivo) e feito seguimento pós-operatório, aos 7, 21, 60 e 180 dias. Os dados foram submetidos à avaliação estatística. RESULTADOS: Não foram observados casos de complicação decorrente do uso do 5-FU em injeção no intraoperatório do pterígio. A taxa de recidiva geral observada aos 180 dias de pós-operatório foi de 35,8%, sendo de 35,7% para os pterígios primários e de 36,4% para os recidivados. CONCLUSÃO: A aplicação do 5-FU no período intraoperatório sob a forma de infiltração subconjuntival é segura. Entretanto, ainda resulta em altas taxas de recidiva e novos estudos devem ser realizados a fim de conhecer a concentração/dose ideal que permitirá menores chances de recidiva da lesão.

Fixação de enxerto cutâneo em malha de espessura total com sutura ou cola de fibrina

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ecballocystis pulvinata Bohlin (Hormotilaceae, Chlorophyta): morfologia externa e ocorrência em ambientes lóticos da região Sudeste do Brasil

O gênero Ecballocystis foi proposto por Bohlin, em 1897, a partir da descrição da espécie Ecballocystis pulvinata coletada no Brasil (Rio Grande do Sul). O gênero é considerado de rara ocorrência no mundo. No Brasil, E. pulvinata tem sido citada com maior frequência. Diversos espécimes de E. pulvinata provenientes de diferentes riachos foram avaliados taxonomicamente por meio de estudos morfométricos e verificou-se a ocorrência desta espécie em relação a algumas variáveis ambientais. Considerável amplitude morfológica foi constada nas populações avaliadas, provavelmente relacionada às variações do ambiente. A característica que mais variou foi o número de cloroplastos, sugerindo que este atributo não seja um caráter taxonômico confiável. A presença de E. pulvinata em riachos de leito rochoso e correnteza rápida (38,5-75,0 cm s-1), mostra que, provavelmente, este tipo de hábitat seja o mais adequado ao seu desenvolvimento.

Controle de erosão em estradas rurais não pavimentadas, utilizando sistema de terraceamento com gradiente associado a bacias de captação

The agricultural unpaved roads are important road structures for the economic and social agricultural development of the Nation, and the erosion provoked by rain water in the road bed and sides are closely related to bad draining, one of the main factors for their degradation. In order to make the draining system adequate, it is necessary to know about erodibility, infiltration capacity of water in the ground and adoption of mechanical slope abatement with grid elevation and water interception. This study presents drainage model through the construction of terraces with gradient transversally associated to the capitation basin in abruptic red dystrophic argisol soils, medium sandy texture, based on slops abatement techniques, elevating the road bed and deviating flow. The grain sized fractions of this ground (sand, silt, clay) and the aggregate stability indicated that this ground, under anthropic action, presents erosive processes resulting in superficial draining with ground hauling, sanding sources and courses of water situation below roads, providing great environmental impacts in the hydric bodies. The reduction of erosion problems in these unpaved roads is in the adoption of measures that intercept waters from the draining of their stream bed itself, as well as pluvial waters comings from adjacent areas of contribution, that are collected and conducted to inlaid terraces and capitation basis.

Comparação da estabilização da manipueira com calcário e hidróxido de sódio na fase acidogênica da biodigestão anaeróbia e uso do biofertilizante

Pós-graduação em Agronomia (Energia na Agricultura) - FCA