The FML (Fucose Mannose Ligand) of Leishmania donovani: a new tool in diagnosis, prognosis, transfusional control and vaccination against human kala-azar

The Fucose-Mannose Ligand (FML) of Leishmania donovani is a complex glycoproteic fraction. Its potential use as a tool for diagnosis of human visceral leishmaniasis was tested with human sera from Natal, Rio Grande do Norte, Brazil. The FML-ELISA test, showed 100% sensitivity and 96% specificity, identifying patients with overt kala-azar (p < 0.001, when compared to normal sera), and subjects with subclinical infection. More than 20% apparently healthy subjects with positive reaction to FML developed overt kala-azar during the following 10 months. In the screening of human blood donnors, a prevalence of 5% of sororeactive subjects was detected, attaining 17% in a single day. The GP36 glycoprotein of FHL is specifically reconized by human kala-azar sera. The immunoprotective effect of FML on experimental L. donovanii infection was tested in swiss albino mice. The protection scheemes included three weekly doses of FML, supplemented or not with saponin by the subcutaneous or intraperitoneal routes and challenge with 2x 10(7) amastigotes of Leishmania donovani. An enhancement of 80.0 % in antibody response (p<0.001) and reduction of 85.5 % parasite liver burden (p<0.001) was detected in animals immunized with FML saponin, unrespectivety of the immunization route.

Use of crude, FML and rK39 antigens in ELISA to detect anti-Leishmania spp. antibodies in Felis catus

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

Saponins, IL12 and BCG adjuvant in the FML-vaccine formulation against murine visceral leishmaniasis

The FML antigen of Leishmania donovani, in combination with either Riedel de Haen (R), QuilA, QS21 saponins, IL12 or BCG, was used in vaccination of an outbred murine model against visceral leishmaniasis (VL). Significant and specific increases in anti-FML IgG and IgM responses were detected for all adjuvants, and in anti-FML IgG1, IgG2a and IgG2b and delayed type of hypersensitivity to L. donovani lysate (DTH), only for all saponins and IL12. The QS21-FML and QuilA-FML groups achieved the highest IgG2a response. QuilA-FML developed the strongest DTH and QS21-FML animals showed the highest serum IFN-gamma concentrations. The reduction of parasitic load in the liver in response to each FML-vaccine formulation was: 52% (P < 0.025) for BCG-FML, 73% (P < 0.005) for R-FML, 93% (P < 0.005) for QuilA-FML and 79.2% (P < 0.025) for QS21-FML treated animals, respectively. Protection was specific for R-FML and QS21-FML while the QuilA saponin treatment itself induced 69% of LDU reduction. The FML-saponin vaccines promote significant, specific and strong protective effects against murine visceral leishmaniasis. BCG-FML induced minor and non-specific protection while IL 12-FML, although enhancing the specific antibody and IDR response, failed to reduce the parasitic load of infected animals. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Immunotherapy against murine experimental visceral leishmaniasis with the FML-vaccine

The fucose mannose ligand (Leishmania donovani FML)-saponin vaccine has earlier shown its immunoprophylactic potential against visceral leishmaniasis in the CB hamster (87.7% of parasite load reduction), Balb/c (84.4%) and Swiss albino mouse (85-93%) models. In this investigation its specific immunotherapeutic efficacy against L. donovani infection in Balb/c mice was studied. The effects of vaccine treatment on the Immoral response, delayed type of hypersensitivity to promastigote lysate (DTH), cytokine levels in sera and reduction of the liver parasitic load of L. donovani infected mice, were examined. The types and subtypes of anti-FML antibodies increased significantly in the vaccinees over the saline and saponin controls. As expected for a saponin vaccine, the highest ratios were found in relation to IgG1, IgG2a and IgG2b (4.4, 5 and 2.5, respectively). The DTH response and the in vitro ganglion cell proliferative response against FML antigen were also significantly higher than controls (P < 0.005). Concomitantly, an impressive and specific decrease of liver parasitic burden was detected only in vaccine-treated animals (94.7%). Our results indicate that the therapeutic FML-vaccine has a potent effect on modulation of the murine infection leading to the reduction of parasitic load and signs of disease, being a new potential tool in the therapy and control of visceral leishmaniasis. (C) 2003 Elsevier Ltd. All rights reserved.

The Fabrication of polyethylene FML field seams /

"September 1989."

Adaptive Immunity against Leishmania Nucleoside Hydrolase Maps Its C-Terminal Domain as the Target of the CD4+ T Cell-Driven Protective Response

Nucleoside hydrolases (NHs) show homology among parasite protozoa, fungi and bacteria. They are vital protagonists in the establishment of early infection and, therefore, are excellent candidates for the pathogen recognition by adaptive immune responses. Immune protection against NHs would prevent disease at the early infection of several pathogens. We have identified the domain of the NH of L. donovani (NH36) responsible for its immunogenicity and protective efficacy against murine visceral leishmaniasis (VL). Using recombinant generated peptides covering the whole NH36 sequence and saponin we demonstrate that protection against L. chagasi is related to its C-terminal domain (amino-acids 199-314) and is mediated mainly by a CD4+ T cell driven response with a lower contribution of CD8+ T cells. Immunization with this peptide exceeds in 36.73 +/- 12.33% the protective response induced by the cognate NH36 protein. Increases in IgM, IgG2a, IgG1 and IgG2b antibodies, CD4+ T cell proportions, IFN-gamma secretion, ratios of IFN-gamma/IL-10 producing CD4+ and CD8+ T cells and percents of antibody binding inhibition by synthetic predicted epitopes were detected in F3 vaccinated mice. The increases in DTH and in ratios of TNF alpha/IL-10 CD4+ producing cells were however the strong correlates of protection which was confirmed by in vivo depletion with monoclonal antibodies, algorithm predicted CD4 and CD8 epitopes and a pronounced decrease in parasite load (90.5-88.23%; p = 0.011) that was long-lasting. No decrease in parasite load was detected after vaccination with the N-domain of NH36, in spite of the induction of IFN-gamma/IL-10 expression by CD4+ T cells after challenge. Both peptides reduced the size of footpad lesions, but only the C-domain reduced the parasite load of mice challenged with L. amazonensis. The identification of the target of the immune response to NH36 represents a basis for the rationale development of a bivalent vaccine against leishmaniasis and for multivalent vaccines against NHs-dependent pathogens.

Diminished CD4+/CD25+T cell and increased IFN-gamma levels occur in dogs vaccinated with Leishmune (R) in an endemic area for visceral leishmaniasis

The Leishmune (R) vaccine has been used in endemic areas to prevent canine visceral leishmaniasis in Brazil, but cytokine production induced by vaccination has rarely been investigated in dogs. This study aimed to evaluate the immune response of dogs vaccinated with Leishmune FML vaccine (Fort Dodge) against total antigen of Leishmania (Leishmania) chagasi (TAg) and FML. Twenty healthy dogs from Aracatuba, Sao Paulo, Brazil, an endemic leishmaniasis area, received three consecutive subcutaneous injection of Leishmune vaccine at 21-day intervals. PBMC were isolated before and 10 days after completing vaccination and lymphoproliferative response and antibody production against FML or total promastigote antigen were tested. Cytokines IFN-gamma, IL-4 and TNF-alpha were measured in culture supernatant and CD4+/CD25+ and CD8+/CD25+ T cell presence was determined. Analysis of the data indicated that the vaccine conferred humoral responses (100%) against both antigens and cellular immunity to FML (85%) and total antigen (80%), the supernatant of cultured cells stimulated with TAg and FML showed an increase in IFN-gamma (P < 0.05), and the vaccine reduced CD4+/CD25+ T cell presence compared to that observed before vaccination. These responses may constitute part of the immune mechanism induced by Leishmune. (C) 2010 Elsevier B.V. All rights reserved.

Projecto de tratamento do Espólio do cientista e Professor José Francisco David Ferreira - Enquadramento, ponto de situação problemas e soluções

O objectivo deste trabalho, relaciona-­‐se com o levantamento de informação escrita relativa ao espólio documental deixado por José Francisco David Ferreira (JFDF), Cientista e Professor Catedrático da Faculdade de Medicina de Lisboa (FML) nascido em 1929 e falecido em 2012. Versa a descrição do processo e da metodologia utilizada, para o projecto em curso, de tratamento do espólio do Cientista e Professor José Francisco David Ferreira. Está enquadrado nos Espólios de Personalidades & Arquivos Científicos em História Contemporânea da Faculdade de Ciências Sociais e Humanas da Universidade Nova de Lisboa.

Malolactic fermentation in winemaking: evaluation of operational parameters

Dissertação de mestrado em Bioengenharia

Histopatologia do saco herniário da hérnia inguinal indireta e do peritônio parietal em adultos e crianças: estudo qualitativo da sua musculatura lisa

Estudo prospectivo, realizado entre abril de 1993 e julho de 1995, com o objetivo de estudar a histopatologia do saco herniário de hérnias inguinais indiretas do adulto e criança, no sentido de verificar a existência de musculatura lisa, sua incidência, apresentação morfológica e comparar com biópsias aleatórias do peritônio parietal. Os pacientes foram divididos em Grupo (1) com 123 pacientes, nos quais foram estudados os sacos herniários, e Grupo (2) constituído de 63 pacientes, nos quais foram realizadas biópsias da serosa peritoneal da cavidade abdominal. Verificou-se que fibras de músculo liso (FML) estiveram presentes em 65,4% dos 133 sacos herniários (dez pacientes com hérnia bilateral), estando presentes, também, em 19,04% dos espécimes da cavidade abdominal. Através dos testes do Qui-quadrado e t de Student, foi avaliada a associação entre a presença de FML com as variáveis categóricas (sexo, cor e lado da hérnia) e as variáveis contínuas (idade dos pacientes, comprimento e espessura do saco herniário). Os resultados mostraram que o sexo feminino apresenta uma maior incidência de FML (p=0,004) e a razão das chances (O.R.) demonstra que os pacientes desse mesmo sexo têm 5,46 vezes mais possibilidades de possuir FML nos sacos herniários. Assim, concluem que as FML são predominantes no peritônio parietal do abdome inferior e que existe, também, uma quantidade maior de FML no peritônio do saco herniário inguinal quando comparado com o peritônio parietal.

Histologia dos sacos herniários das hérnias inguinais indiretas, diretas, recidivadas e encarceradas em adultos e crianças: identificação de fibras musculares lisas

OBJETIVO: Pesquisar a presença de fibras de músculo liso (FML) nos sacos peritoneais das hérnias indiretas, diretas, recidivadas e encarceradas e estudar a influência do sexo, cor e idade dos pacientes, bem como a região do saco herniário, largura, comprimento e espessura da biópsia coletada. MÉTODO: Foram obtidos 252 sacos herniários no período de fevereiro de 1999 a dezembro de 2000 e encaminhados para o estudo histopatológico através da coloração por hematoxilina-eosina e tricrômico de Gomori. A idade variou entre um mês a 87 anos com média de 42,3 anos e desvio-padrão de 22,5 anos. RESULTADOS: Foi utilizado o teste do Qui-quadrado e observada FML em 76,5% dos pacientes com hérnia indireta, 55,9% direta, 46,4% encarcerada e 68,7% recidivada. No estudo global foram encontradas FML em 67,9% dos espécimes com maior incidência na porção proximal do saco herniário (53,2%) e em pacientes melanodérmicos (75%). As FML estiveram presentes em maior freqüência no lado direito (67,7%) e no sexo feminino (73,3%). Do total de 252 amostras de sacos herniários examinados, foi encontrada FML em 171 biópsias, e esse achado foi menos freqüente nas hérnias diretas e encarceradas quando comparadas com as indiretas e recidivadas. CONCLUSÕES: Como hipótese, a presença de FML na parede do saco herniário pode representar um reforço tecidual no sentido de dificultar o crescimento do saco peritoneal, comportando-se como fator de resistência elástica e dinâmica à expansão da hérnia. Por outro lado, pode também significar uma formação aberrante ou a persistência de uma estrutura que deveria regredir ou mesmo desaparecer durante o desenvolvimento normal.

Selecci??n de modelos anidados para datos longitudinales usando criterios de informaci??n y la estrategia de ajuste condicional

Resumen tomado de la publicaci??n

Evaluación psicopedagógica de un programa para la enseñanza deportiva de vela.

Construir un instrumento psicométrico útil y válido para evaluar la eficacia de un programa de enseñanza alternativa basado en los contenidos del SOE, y para evaluar la capacidad adquirida por sujetos que han participado en programas de iniciación. Redefinir el término iniciación desde una perspectiva psicológica. Determinar un modelo de procedimiento de enseñanza y evaluación útil de actividad de ocio. La muestra la componían 117 individuos, alumnos de una escuela de vela. La investigación se articula en dos bloques; en el primero de ellos se plantea la iniciación a la vela desde una perspectiva histórica, sociológica y psicológica; desde esta última perspectiva se determinan los contenidos y habilidades que el sujeto iniciado debe poseer. A continuación se elabora un programa de enseñanza y evaluación basado en las teorías de Piagelin, inscritas dentro de la perspectiva genético-evolutiva de J. Piaget. En el segundo bloque se comprueba experimentalmente la validez del método mediante los instrumentos seleccionados y se analizan los datos desde las perspectivas psicométricas siguientes: test norma de grupo, test referidos al criterio y desde la teoría de respuesta de ítems. Test del lápiz y papel con ítems gráficos. Software para el análisis de ítems siguientes: UNIVAC, BICAL, ANCILLES, CALFIT, ITEMAN, BIG JIFFY, NOHARM, CREDIT, FML, BILOG, DISLOG, LOGISTY, TESTFACT, MICROCAT, MULTILOS, MIRTE, S/N, METRIX. Según software. Se confirma la importancia del planteamiento psicológico de un modelo de aprendizaje de la vela. Se observa que la excesiva tecnificación puede repercutir negativamente en la motivación aunque los resultados sean más positivos.

Elastic properties of hygrothermally conditioned glare laminate

The influences of hygrothermal conditioning on mechanical properties of a fiber/metal laminate (FML) have been investigated by tensile and compression tests. The environmental action, such as high moisture concentration, high temperatures, corrosive fluids or ultraviolet radiation (UV), can affect the performance of advanced composites during service. In the present work, the results show that for the glass fiber/epoxy composites tensile and compression values decrease after hygrothermal conditioning. However, no changes on mechanical properties (tensile and compression strength) are observed for the Glare laminate, regardless the hygrothermal conditioning. (C) 2006 Elsevier Ltd. All rights reserved.

Hygrothermal effects on damping behavior of metal/glass fiber/epoxy hybrid composites

Continuous fiber/metal laminates (FML) offer significant improvements over current available materials for aircraft structures due to their excellent fatigue endurance and low density. Glass fibers/epoxy laminae and aluminum foil (Glare) are commonly used to obtain these hybrid composites. The environmental factors can limit the applications of composites by deteriorating the mechanical properties during service. Usually, epoxy resins absorb moisture when exposed to humid environments and metals are prone to surface corrosion. Therefore, the combination of the two materials in Glare (polymeric composite and metal). can lead to differences that often turn out to be beneficial in terms of mechanical properties and resistance to environmental influences. In this work. The viscoelastic properties. such as storage modulus (E') and loss modulus (E'), were obtained for glass fiber/epoxy composite, aluminum 2024-T3 alloy and for a glass fiber/epoxy/aluminum laminate (Glare). It was found that the glass fiber/epoxy (G/E) composites decrease the E' modulus during hygrothermal conditioning up to saturation point (6 weeks). However, for Glare laminates the E' modulus remains unchanged (49GPa) during the cycle of hygrothermal conditioning. The outer aluminum sheets in the Glare laminate shield the G/E composite laminae from moisture absorption. which in turn prevent, in a certain extent, the material from hygrothermal degradation effects. (c) 2005 Elsevier B.V. All rights reserved.