Inhibition of enzyme activity of Rhipicephalus (Boophilus) microplus triosephosphate isomerase and BME26 cell growth by monoclonal antibodies.

In the present work, we produced two monoclonal antibodies (BrBm37 and BrBm38) and tested their action against the triosephosphate isomerase of Rhipicephalus (Boophilus) microplus (RmTIM). These antibodies recognize epitopes on both the native and recombinant forms of the protein. rRmTIM inhibition  by BrBm37 was up to 85% whereas that of BrBrm38 was 98%, depending on the antibody-enzyme ratio. RmTIM activity was lower in ovarian, gut, and fat body tissue extracts treated with BrBm37 or BrBm38 mAbs. The proliferation of the embryonic tick cell line (BME26) was inhibited by BrBm37 and BrBm38 mAbs. In summary, the results reveal that it is possible to interfere with the RmTIM function using antibodies, even in intact cells.

Anti-tick monoclonal antibody applied by artificial capillary feeding in Rhipicephalus (Boophilus) microplus females.

The tick Rhipicephalus microplus is an ectoparasite harmful to livestock, a vector of disease agents that affects meat and milk production. However, resistance to acaricides reflects the need for alternative tick control methods, among which vaccines have gained increasing relevance. In this scenario, monoclonal antibodies can be used to identify and characterize antigens that can be used as vaccine immunogens. Capillary tube artificial feeding of partially engorged R. microplus females with monoclonal antibodies against proteins from the gut of tick were used to test the effects of immunoglobulins in the physiology of the parasite. The results of artificial feeding showed that female ticks over 25mg and under 60 mg in weight performed better in the artificial feeding process, with a 94-168% weight increase after 24h of feeding. Results showed that artificial feeding of ticks proved to be a viable technique to study the effects of antibodies or drugs in the physiology of the parasite. One monoclonal antibody (BrBm2) induced decreased oviposition. Moreover, the antigen recognized by BrBm2 was identified as a 27-kDa protein and immunolabeled on digestive vesicles membranes of digestive cells of partially and fully engorged females.

Monoclonal antibodies against peptidorhamnomannans of Scedosporium apiospermum enhance the pathogenicity of the fungus.

Scedosporium apiospermum is part of the Pseudallescheria-Scedosporium complex. Peptidorhamnomannans (PRMs) are cell wall glycopeptides present in some fungi, and their structures have been characterized in S. apiospermum, S. prolificans and Sporothrix schenckii. Prior work shows that PRMs can interact with host cells and that the glycopeptides are antigenic. In the present study, three monoclonal antibodies (mAbs, IgG1) to S. apiospermum derived PRM were generated and their effects on S. apiospermum were examined in vitro and in vivo. The mAbs recognized a carbohydrate epitope on PRM. In culture, addition of the PRM mAbs increased S. apiospermum conidia germination and reduced conidial phagocytosis by J774.16 macrophages. In a murine infection model, mice treated with antibodies to PRM died prior to control animals. Thus, PRM is involved in morphogenesis and the binding of this glycopeptide by mAbs enhanced the virulence of the fungus. Further insights into the effects of these glycopeptides on the pathobiology of S. apiospermum may lead to new avenues for preventing and treating scedosporiosis.

Monoclonal antibody to fungal glucosylceramide protects mice against lethal Cryptococcus neoformans infection.

Glucosylceramides (GlcCer) are involved in the regulation of Cryptococcus neoformans virulence. In the present study, we demonstrate that passive immunization with a monoclonal antibody to GlcCer significantly reduces host inflammation and prolongs the survival of mice lethally infected with C. neoformans, revealing a potential therapeutic strategy to control cryptococcosis.

Vaccination of bovines with recombinant Boophilus Yolk pro-Cathepsin.

Boophilus Yolk pro-Cathepsin (BYC) is an aspartic proteinase found in Boophilus microplus eggs that is involved in the embryogenesis and has been tested as antigen to compose an anti-tick vaccine. The vaccine potential of a recombinant BYC expressed in Escherichia coli (rBYC) was investigated. rBYC was purified and used to immunize Hereford cattle. The sera of bovines immunized with rBYC recognized the native BYC with a titer ranging from 125 to 4000. Furthermore, immunized bovines challenged with 20,000 larvae presented an overall protection of 25.24%. The partial protection obtained against B. microplus infestation with the recombinant protein immunization was similar to the already described for native BYC immunization.

Teratoma mediastínico: causa poco frecuente de dificultad respiratoria aguda en lactantes / Mediastinal teratoma: an unusual cause of acute respiratory distress in infants

RESUMEN Se presenta un lactante de 5 meses de edad con síndrome de dificultad respiratoria de evolución tórpida. Se solicitó radiografía de tórax frente y perfil detectándose una masa mediastínica de gran tamaño y estructura heterogénea. Se completó el estudio con ultrasonografía y tomografía computada con contraste de tórax y sin él; éste último método fue el que orientó el diagnóstico prequirúrgico de teratoma. Los marcadores de actividad tumoral fueron negativos. Se practicó la resección quirúrgica completa del tumor y parte del timo normal.La evolución posquirúrgica del paciente fue favorable.La anatomía patológica confirmó el diagnóstico de teratoma inmaduro. El objetivo ha sido presentar una causa muy poco frecuente de dificultad respiratoria en lactantes, así como poner de manifiesto la escasa inespecífica sintomatología(AU)

Teratoma mediastínico: causa poco frecuente de dificultad respiratoria aguda en lactantes / Mediastinal teratoma: an unusual cause of acute respiratory distress in infants

RESUMEN Se presenta un lactante de 5 meses de edad con síndrome de dificultad respiratoria de evolución tórpida. Se solicitó radiografía de tórax frente y perfil detectándose una masa mediastínica de gran tamaño y estructura heterogénea. Se completó el estudio con ultrasonografía y tomografía computada con contraste de tórax y sin él; éste último método fue el que orientó el diagnóstico prequirúrgico de teratoma. Los marcadores de actividad tumoral fueron negativos. Se practicó la resección quirúrgica completa del tumor y parte del timo normal.La evolución posquirúrgica del paciente fue favorable.La anatomía patológica confirmó el diagnóstico de teratoma inmaduro. El objetivo ha sido presentar una causa muy poco frecuente de dificultad respiratoria en lactantes, así como poner de manifiesto la escasa inespecífica sintomatología

Purification and antigenicity of two recombinant forms of Boophilus microplus yolk pro-cathepsin expressed in inclusion bodies.

The tick Boophilus microplus is a bovine ectoparasite present in tropical and subtropical areas of the world and the use of vaccines is a promising method for tick control. BYC is an aspartic proteinase found in eggs that is involved in the embryogenesis of B. microplus and was proposed as an important antigen in the development of an anti-tick vaccine. The cDNA of BYC was amplified by PCR and cloned for expression in two forms with and without thioredoxin fusion protein (Trx), coding recombinant proteins named rBYC-Trx and rBYC, respectively. Expression, solubility, and yields of the two forms were analyzed. The recombinant proteins were expressed in inclusion bodies (IBs) and three denaturant agents (N-lauroyl sarcosine, guanidine hydrochloride, and urea) were tested for IBs solubilization. The N-lauroyl sarcosine solubilized 90.4 and 92.4% of rBYC-Trx and rBYC IBs, respectively, and was the most efficient denaturant. Two recombinant forms were affinity-purified by Ni2+-Sepharose under denaturing conditions. After dialysis, the yield of soluble protein was 84.1% for r-BYC-Trx and 5.9% for rBYC. These proteins were immune-reactive against sera from rabbit, mouse, and bovine previously immunized with native BYC, which confirms the antigenicity of the recombinant BYCs expressed in the Escherichia coli system.

Representação social, lazer e mulheres portadoras de cegueira congênita / Social representation, leisure and women with congenital blindness

O presente estudo tem como objetivo compreender as representações sociais das mulheres portadoras de cegueira congênita sobre sua deficiência e as interferências no lazer. Trata-se de um estudo de natureza qualitativa, realizado com dezessete mulheres portadoras de cegueira congênita que freqüentaram o Centro de Apoio Pedagógico ao Deficiente Visual (CAP) em Salvador-Bahia, em 2004. Para a coleta de dados, utilizou-se a entrevista semi-estruturada. Estas foram analisadas e interpretadas através da análise de conteúdo. A partir da análise, apresenta-se o núcleo de significação: compreensão e sentimentos sobre a deficiência e a interferência no desenvolvimento do lazer. As representações sociais destas mulheres assinalam um pensar e sentir apoiados por uma visão paternalista, assistencialista e segregadora que a sociedade apresenta, fundamentada no que elas denominam de doença social, ou seja, a falta de informação e um discurso de identidade em que manifestam o desejo de serem reconhecidas como pessoas normais, pensantes e afetivas. Foi possível apreender que as mulheres portadoras de cegueira congênita arcam o ônus da doença social, entretanto em suas representações e sentimentos apontam subsídios para a busca da superação desta construção histórica arraigada nas representações dos grupos sociais no que se refere à mulher portadora de cegueira congênita e à interferência no lazer.

Purification and ultrastructural localization of a copper-zinc superoxide dismutase (CuZnSOD) from the entomopathogenic and acaricide fungus Metarhizium anisopliae.

The entomopathogenic fungus Metarhizium anisopliae contains three superoxide dismutases. One of these enzymes was purified and partially characterized as a CuZnSOD. The enzyme has an estimated molecular mass of 30690 Da and a specific activity of 3838.89 Umg(-1). SDS-PAGE and 2D gels show a single band of protein in the fractions eluted from the gel filtration column with a molecular mass of 20000 and approximately 15000 Da, respectively, and a pI of 6.0. These results suggest that the native enzyme is a dimer consisting of two subunits. Polyclonal antiserum were raised against purified CuZnSOD and used to determine its subcellular localization by immunoelectron microscopy. M. anisopliae CuZnSOD is present in the cell wall.

A monoclonal antibody to glucosylceramide inhibits the growth of Fonsecaea pedrosoi and enhances the antifungal action of mouse macrophages.

Fungal glucosylceramides (GlcCer) are conserved lipid components in a large variety of pathogenic and non-pathogenic fungal species, but their biological functions are still unclear. Recent studies demonstrate that GlcCer are immunologically active components inducing the production of antifungal antibodies. In this work, a major GlcCer was purified and characterized from mycelial forms of Fonsecaea pedrosoi, the most frequent causative agent of chromoblastomycosis. As determined by fast atom bombardment mass spectrometry (FAB-MS) analysis, the purified molecule was identified as the conserved structure N-2'-hydroxyhexadecanoyl-1-beta-D-glucopyranosyl-9-methyl-4,8-sphingadienine. A monoclonal antibody (Mab) against this structure was used in indirect immunofluorescence with the different morphological stages of F. pedrosoi. Only the surface of young dividing cells was recognized by the antibody. Treatment of F. pedrosoi conidia with the Mab against GlcCer resulted in a clear reduction in fungal growth. In addition, the Mab also enhanced phagocytosis and killing of F. pedrosoi by murine cells. These results suggest that, in F. pedrosoi, GlcCer seem to be cell wall components targeted by antifungal antibodies that directly inhibit fungal development and also enhance macrophage function, supporting the use of monoclonal antibodies to GlcCer as potential tools in antifungal immunotherapy.

Glucosylceramides in Colletotrichum gloeosporioides are involved in the differentiation of conidia into mycelial cells.

Glucosylceramides (GlcCer) were extracted from the plant pathogen Colletotrichum gloeosporioides and purified by several chromatographic steps. By using electrospray ionization mass spectrometry and nuclear magnetic resonance, GlcCer from C. gloeosporioides were identified as N-2'-hydroxyoctadecanoyl-1-beta-D-glucopyranosyl-9-methyl-4,8-sphingadienine and N-2'-hydroxyoctadecenoyl-1-beta-D-glucopyranosyl-9-methyl-4,8-sphingadienine. Monoclonal antibodies against these structures were produced and used as tools for the evaluation of the role of GlcCer in the morphological transition of C. gloeosporioides. In the presence of antibodies to GlcCer, the differentiation of conidia into mycelia was blocked. Since GlcCer is present in several plant pathogens, the inhibitory activity of external ligands recognizing these structures may be applicable in other models of fungal infections.

In vitro segmentation induction of Mesocestoides corti (Cestoda) tetrathyridia.

Mesocestoides corti is a suitable model for studying cestode development because of its ability to reproduce asexually and segment in vitro. The cultured parasite is also capable of sexual differentiation and, probably, reproduction. To establish conditions that increase the efficiency of in vitro M. corti larvae (tetrathyridia) segmentation, we tested the effects of an inducing agent and some physical parameters in cultures. We found that a 5% CO2-95% N2 gas phase, an incubation temperature of 39 C (instead of 37 C), and a 24-hr pretreatment with trypsin (10(5) BAEE/ml, BAEE = Na-benzoil-L-arginine ethyl ester unit of trypsin activity) in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with 20% fetal bovine serum (FBS) are able to increase individually or synergistically the segmentation rate of tetrathyridia. A segmentation rate of up to 100% was achieved on day 4 of culture, when all these conditions were used simultaneously, in comparison with an average rate of 40% obtained not before day 11 in cultures without any inducing treatment. Fetal bovine serum is essential for segmentation, and a concentration of 20% was established as the standard for induction.

Alfa-interferon recombinante no tratamento das hepatites crônicas B e näo-A: resultados preliminares / Recombinant alpha - interferon in the treatment of B and non-A, non-B chronic hepatitis: preliminary results

O interferon-alfa recombinatne (IFN-R) foi administrado a 17 pacientes com hepatite crônica näo-A, näo-B (HCNANB) e com 11 hepatite crônica B (HCB). Apesar da febre sistemática após as primeiras injeçöes (38 a 39 graus Celsius), de outros sintomas como calafrios, dores musculares e cefaléias e das alteraçöes de leucócitos, neutrófilos e por vezes de plaquetas, a tolerância foi considerada satisfatória, sendo os efeitos colaterais reversíveis com o término do tratamento. A eficácia na HCNANB, representada pela normalizaçäo das aminotransferases ou sua reduçäo para níveis inferiores a duas vezes o limite máximo normal, foi observada em quatro de oito pacientes que completaram o tratamento em períodos de três a nove meses. Dos quatro respondedores, um paciente, tratado durante cinco meses, voltou a apresentar elevaçöes das transaminases três meses após o término do tratamento. Por outro lado, em um paciente com HC ativa em fase cirrótica e tratado durante 12 meses observou-se normalizaçäo persistente das aminotransferases, tendo a biópsia de controle revelado acentuada reduçäo do processo inflamatório. A eficácia na HCB, representada pelo desaparecimento do AgHBe e da DNA-polimerase foi observada em três dos sete pacientes que completaram o tratamento. Esses resultados preliminares mostram que o IFN é uma droga promissora, mas sosmente os estudos controlados multicêntricos poderäo estabelecer definitivamente seu valor no tratamento das hepatites crônicas virais

Incidência de sífilis em gestantes / Incidence of syphylis in pregnancy

Realizou-se uma investigaçäo sorológica (VDRL quantitativo e hemaglutinaçäo qualitativa) em 200 gestantes primíparas internadas na Unidade Mista Prof. Barros Lima (Recife), para o diagnóstico da sífilis. As parturientes apresentavam idade entre 14 (dois casos) e 35 anos (10 casos); cor parda em 130 e preta em 70 pacientes; a procedência das mesmas foi em 180 da capital (diversos bairros) e em 20 de cidades do interior do Estado de Pernambuco; a profissäo dominante foi a de doméstica, em 120 gestantes, seguida de cambista, com 30 parturientes; balconista e servente, com 20 casos cada e ambulantes com 10. Em todas foram executados o VDRL quantitativo, sendo positivo em 23 gestantes (11,5%) e nestas efetuamos a hemaglutinaçäo qualitativa, a qual foi positiva em oito gestantes (34,78%)