Potentiation of polarized intestinal Caco-2 cell responsiveness to probiotics complexed with secretory IgA.

The precise mechanisms underlying the interaction between intestinal bacteria and the host epithelium lead to multiple consequences that remain poorly understood at the molecular level. Deciphering such events can provide valuable information as to the mode of action of commensal and probiotic microorganisms in the gastrointestinal environment. Potential roles of such microorganisms along the privileged target represented by the mucosal immune system include maturation prior, during and after weaning, and the reduction of inflammatory reactions in pathogenic conditions. Using human intestinal epithelial Caco-2 cell grown as polarized monolayers, we found that association of a Lactobacillus or a Bifidobacterium with nonspecific secretory IgA (SIgA) enhanced probiotic adhesion by a factor of 3.4-fold or more. Bacteria alone or in complex with SIgA reinforced transepithelial electrical resistance, a phenomenon coupled with increased phosphorylation of tight junction proteins zonula occludens-1 and occludin. In contrast, association with SIgA resulted in both enhanced level of nuclear translocation of NF-κB and production of epithelial polymeric Ig receptor as compared with bacteria alone. Moreover, thymic stromal lymphopoietin production was increased upon exposure to bacteria and further enhanced with SIgA-based complexes, whereas the level of pro-inflammatory epithelial cell mediators remained unaffected. Interestingly, SIgA-mediated potentiation of the Caco-2 cell responsiveness to the two probiotics tested involved Fab-independent interaction with the bacteria. These findings add to the multiple functions of SIgA and underscore a novel role of the antibody in interaction with intestinal bacteria.

La función social de la investigación

Este artículo reflexiona y analiza la función social de la investigación. Evidencia la importancia y complementariedad de los tres ámbitos del conocimiento científico que son: la Tecnología como componente presente en casi la totalidad de investigaciones. La Ciencia, elemento clave y básico para explicar los hechos, datos o acontecimientos que se investigan, así como la Innovación, que es la que garantiza que siga avanzando el conocimiento. El artículo se detiene a analizar los aspectos clave de la función social de la investigación y sus especificidades, tanto en la investigación básica como en la aplicada. Concluye el texto con el análisis de la responsabilidad social de la investigación así como la importancia de la transferencia de los resultados de la misma a la sociedad, animando a la construcción social de una comunidad investigadora que siga trabajando para repercutir socialmente los resultados de sus investigaciones.

Antigen binding to secretory immunoglobulin A results in decreased sensitivity to intestinal proteases and increased binding to cellular Fc receptors.

In intestinal secretions, secretory IgA (SIgA) plays an important sentinel and protective role in the recognition and clearance of enteric pathogens. In addition to serving as a first line of defense, SIgA and SIgA x antigen immune complexes are selectively transported across Peyer's patches to underlying dendritic cells in the mucosa-associated lymphoid tissue, contributing to immune surveillance and immunomodulation. To explain the unexpected transport of immune complexes in face of the large excess of free SIgA in secretions, we postulated that SIgA experiences structural modifications upon antigen binding. To address this issue, we associated specific polymeric IgA and SIgA with antigens of various sizes and complexity (protein toxin, virus, bacterium). Compared with free antibody, we found modified sensitivity of the three antigens assayed after exposure to proteases from intestinal washes. Antigen binding further impacted on the immunoreactivity toward polyclonal antisera specific for the heavy and light chains of the antibody, as a function of the antigen size. These conformational changes promoted binding of the SIgA-based immune complex compared with the free antibody to cellular receptors (Fc alphaRI and polymeric immunoglobulin receptor) expressed on the surface of premyelocytic and epithelial cell lines. These data reveal that antigen recognition by SIgA triggers structural changes that confer to the antibody enhanced receptor binding properties. This identifies immune complexes as particular structural entities integrating the presence of bound antigens and adds to the known function of immune exclusion and mucus anchoring by SIgA.

N-Glycans on secretory component: mediators of the interaction between secretory IgA and gram-positive commensals sustaining intestinal homeostasis.

Human beings live in symbiosis with billions of microorganisms colonizing mucosal surfaces. The understanding of the mechanisms underlying this fine-tuned intestinal balance has made significant processes during the last decades. We have recently demonstrated that the interaction of SIgA with Gram-positive bacteria is essentially based on Fab-independent, glycan-mediated recognition. Results obtained using mouse hybridoma- and colostrum-derived secretory IgA (SIgA) consistently show that N-glycans present on secretory component (SC) play a crucial role in the process. Natural coating may involve specific Gram-positive cell wall components, which may explain selective recognition at the molecular level. More widely, the existence of these complexes is involved in the modulation of intestinal epithelial cell (IEC) responses in vitro and the formation of intestinal biofilms. Thus, SIgA may act as one of the pillars in homeostatic maintenance of the microbiota in the gut, adding yet another facet to its multiple roles in the mucosal environment.

Secretory IgA, a major immunoglobulin in most bovine external secretions.

Bovine secretory IgA (SIgA), recently identified in colostrum, was shown to be homologous to human SIgA by immunologic cross-reaction. A quantitative study indicated that bovine SIgA, a minor component of colostrum, is a major immunoglobulin in most other external secretions including saliva, spermatic fluid, lacrimal, nasal and gastrointestinal secretions. SIgA was isolated from saliva. The free form of secretory component was found to be abundant in milk. A normal lactating cow produces about 1.2 g of this protein per day. Two forms of IgA were identified in serum: a normal serum IgA with no secretory antigenic determinant, and a small amount of SIgA. In vitro synthesis of SIgA by the salivary gland was studied by tissue cultures with incorporation of labeled amino acids.

Recognition of Gram-positive Intestinal Bacteria by Hybridoma- and Colostrum-derived Secretory Immunoglobulin A Is Mediated by Carbohydrates.

Humans live in symbiosis with 10(14) commensal bacteria among which >99% resides in their gastrointestinal tract. The molecular bases pertaining to the interaction between mucosal secretory IgA (SIgA) and bacteria residing in the intestine are not known. Previous studies have demonstrated that commensals are naturally coated by SIgA in the gut lumen. Thus, understanding how natural SIgA interacts with commensal bacteria can provide new clues on its multiple functions at mucosal surfaces. Using fluorescently labeled, nonspecific SIgA or secretory component (SC), we visualized by confocal microscopy the interaction with various commensal bacteria, including Lactobacillus, Bifidobacteria, Escherichia coli, and Bacteroides strains. These experiments revealed that the interaction between SIgA and commensal bacteria involves Fab- and Fc-independent structural motifs, featuring SC as a crucial partner. Removal of glycans present on free SC or bound in SIgA resulted in a drastic drop in the interaction with Gram-positive bacteria, indicating the essential role of carbohydrates in the process. In contrast, poor binding of Gram-positive bacteria by control IgG was observed. The interaction with Gram-negative bacteria was preserved whatever the molecular form of protein partner used, suggesting the involvement of different binding motifs. Purified SIgA and SC from either mouse hybridoma cells or human colostrum exhibited identical patterns of recognition for Gram-positive bacteria, emphasizing conserved plasticity between species. Thus, sugar-mediated binding of commensals by SIgA highlights the currently underappreciated role of glycans in mediating the interaction between a highly diverse microbiota and the mucosal immune system.

Secretory IgA: an actor of the interplay between the commensal flora and the intestinal immune system?

In the gastro-intestinal tract,Peyers patches have been describedas a major inductive site for mucosalsecretory IgA (SIgA) responses directedagainst pathogens. The classicalview is that SIgAserves as the firstline of defense against microorganismsby agglutining potential invadersand faciliting their clearance byperistaltic and mucociliary movements,a mechanism called immuneexclusion. Our laboratory has shownthat SIgA is not only able to be"retrotransported" into Peyers patchesvia the associated M cells, but also todeliver sizeable cargos in the form ofSIgA-based immune complexes, resultingin the onset of non-inflammatorytype of responses. Such a novelfunction raises the question of thepossible role of mucosal SIgA in theinterplay with commensal bacteriaand the contribution of the antibody inbacterial homeostasis. To address thisquestion, Lactobacillus rhamnosus(LPR) was administered into a mouseligated loop comprising a Peyerspatch, in association or not with SIgA.The fate of fluorescently labelled bacteriawas followed by laser scanningconfocal microscopy at different incubationtimes. After 2 hours of incubationin the loop, LPR bacteria arefound more abundantly in thesubepithelial dome (SED) regionwhen they are coated with SIgA thanLPR administered alone despite theyare absent from neighboring villi.Herein, it is shown that this mechanismof entry involves M cells inPeyers pathes. After their sampling byM cells, bacteria are engulfed by thedendritic cells of the subjacent SEDregion. Interestingly, LPR bacteriaare found coated by the endogenousnatural SIgA present in mice intestinalsecretions, confirming the requirementof SIgA for this type of entry.The subsequent effect on the maturationof dendritic cells after interactionwith LPR was investigated in vitroin presence or not of SIgA by measuringthe expression of CD40, CD80and CD86 surface markers with flowcytometry analyses. Results show thatDCs respond in the same way in presenceof SIgA than with LPR bacteriaalone, indicating that SIgA does notmodulate the interaction betweenDCs and bacteria in this context. Thiswork gives new evidences about theinvolvement of SIgA in the mechanismby which the intestinal immunesystem permanently checks the contentof the intestine.

Naevus naevocellulaire achrome palpébral prenant l'aspect d'un carcinome basocellulaire [Nevus mimicking a basal cell carcinoma of the eyelid].

A clinicopathologic case of a 41-year-old female patient exhibited a single cutaneous tumor at the inner part of the free margin of the inferior left eyelid. It was a pink, fleshy, and nodular well-circumscribed exophytic mass with thin vessels on its surface. Experienced already for 20 years, this lesion had been observed 6 years before and has not exhibited much change since then. However, its clinical appearance argued for a possible small basal cell carcinoma, which had grown over the inferior left lachrymal duct. After surgical removal, histopathology showed that the tumor was an amelanotic dermal nevus. No disturbance of lachrymal drainage was observed after surgery. This case shows that nodular amelanotic tumors of the eyelid, even when located on the inner segment of the eyelid, may be a nevus.

Escuela y minorías. Inmigrantes y gitanos ante el modelo educativo español

Entre los grupos anteriores existe un elevado interés por la conservación de la cultura propia y la tradición. Cada uno adopta diferentes estrategias, ante una sociedad que no siente como propia, según los objetivos que tiene para sus hijos. De esta forma encontramos desde a los que no tienen ningún interés por convivir escolarmente con otras culturas, segregacionistas, a aquellos que quieren mantener la tradición y la cultura con los mínimos cambios posibles y que, a la vez, no creen que deba ser conocido por otras personas que no pertenecen a su colectivo, tradicionalistas, hasta a aquellos que a pesar de querer mantener sus particularidades las minimizan, mimetizan, a cambio de movilidad social, asimilacionistas. Pero, cuando se les valora socialmente su cultura y su presencia en la escuela se muestran a favor, pensando que a la larga esta presencia y aceptación puede evitar que sea necesario esconder su adscripción. Otros grupos se han situado dentro del reconocimiento y valoración de su cultura, pero con diferentes niveles de expectativas que han sido corregidas, respecto a lo anteriormente expuesto, por la edad prevista de finalización de los estudios. Desde los conformados con los estudios básicos, e incluso, los que no creen necesaria su finalización y que reivindican la presencia del caló o romaní en la escuela, a los que tienen unas expectativas más elevadas y útiles, manteniendo la cultura aparte y piensan que no debe enseñarse a todos los alumnos. Una formación profesional les resulta más interesante por la mayor practicidad y porque determinadas profesiones que payos y españoles no quieren pueden ser un mercado potencial para ellos. El último grupo situaría su expectativa en un nivel educativo superior, la universidad, considerando que les puede suponer movilidad social, pero reduciendo importancia a la inmediata utilidad que otros han destacado. También su lengua o cultura, a las que no están dispuestos a renunciar, deben seguir estando presentes en todo el proceso y apreciarían que formase parte del currículo escolar. Hemos considerado que la educación intercultural se define como una educación en que la escuela debe formar y fortalecer los valores de igualdad, respeto y pluralismo, en que se reconoce el derecho personal de cada alumno a recibir una educación diferenciada cuidando su identidad , en que se reconocen las culturas y lenguas, en que se intentan superar los estereotipos y prejuicios, etcétera. Este modelo de educación puede suponer una respuesta a unas demandas de las minorías que la escuela no con sigue satisfacer. En definitiva, una escuela que siga el modelo intercultural puede igualar expectativas y evitar que algunos piensen que la escuela sirve para poco y , además, interfiere. Pero, esto también dependerá de las miras que se tenga del futuro laboral que se quiere para sus hijos, si la idea es reproducir muchas de las ocupaciones que actualmente realizan o roles que actualmente asumen, por ejemplo según el género, difícilmente el nivel educativo deseado se incrementará.. A partir de todo lo expuesto es necesario aproximar la escuela a las minorías étnicas con todos los medios necesarios y buscar otras estrategias de captación, de seducción, de estas minorías y sobre todo de los gitanos para que crean que la escuela es también su escuela.

Controle de qualidade e artefatos em mamografia

A mamografia, na atualidade, é o método mais efetivo de diagnóstico precoce do câncer de mama. Um exame com alto padrão de qualidade pode visualizar, em 85% a 90% dos casos, um tumor com mais de dois anos de antecedência de ocorrer acometimento ganglionar, em mulheres com mais de 50 anos de idade. A diferença radiográfica entre o tecido normal e o doente é extremamente tênue, logo, a alta qualidade do exame é indispensável para alcançar resolução de alto contraste que permita essa diferenciação. Para alcançar alto padrão é imperativo que o exame mamográfico siga protocolos rígidos e pré-estabelecidos. Os artefatos são defeitos no processamento do filme que comprometem o resultado final da imagem, podendo resultar em informações perdidas ou mascaradas. Há numerosos tipos de artefatos derivados de diversas fontes na aquisição da imagem, como o processador, o desempenho do técnico de radiologia, o mamógrafo ou o paciente, todos resultando na degradação da imagem obtida. O presente artigo tem o objetivo de revisar métodos eficazes no controle de qualidade do exame mamográfico e analisar os artefatos mais importantes na prática diária, com ilustrações e dicas de como evitá-los.

Circadian variation of salivary immunoglobin A, alpha-amylase activity and mood in response to repeated double-poling sprints in hypoxia.

PURPOSE: To assess the circadian variations in salivary immunoglobin A (sIgA) and alpha-amylase activity (sAA), biomarkers of mucosal immune function, together with mood during 2 weeks of repeated sprint training in hypoxia (RSH) and normoxia (RSN). METHODS: Over a 2-week period, 17 competitive cross-country skiers performed six training sessions, each consisting of four sets of five 10-s bouts of all-out double-poling under either normobaric hypoxia (FiO2: 13.8 %, 3000 m) or normoxia. The levels of sIgA and sAA activity and mood were determined five times during each of the first (T1) and sixth (T6) days of training, as well as during days preceding (baseline) and after the training intervention (follow-up). RESULTS: With RSH, sIgA was higher on T6 than T1 (P = 0.049), and sAA was increased on days T1, T6, and during the follow-up (P < 0.01). With RSN, sIgA remained unchanged and sAA was elevated on day T1 only (P = 0.04). Similarly, the RSH group demonstrated reduced mood on days T1, T6, and during the follow-up, while mood was lowered only on T1 with RSN (P < 0.01). CONCLUSIONS: The circadian variation of sIgA and sAA activity, biomarkers of mucosal immune function, as well as mood were similar on the first day of training when repeated double-poling sprints were performed with or without hypoxia. Only with RSH did the levels of sIgA and sAA activity rise with time, becoming maximal after six training sessions, when mood was still lowered. Therefore, six sessions of RSH reduced mood, but did not impair mucosal immune function.

Autoevaluación: Fisiopatología del metabolismo

La respiración, la circulación sanguínea, la contracción de un músculo o el mero pensamiento son fenómenos que obligan a que nuestro organismo esté en constante cambio, produciendo y consumiendo energía. La palabra metabolismo deriva del griego metabolé y significa"cambio". Entendemos como metabolismo a todo el conjunto de reacciones bioquímicas y procesos físicos que ocurren en la célula y en el organismo. Todos estos procesos metabólicos tienen lugar en dos fases. Una fase, llamada anabolismo, en la que se consume energía para transformar moléculas pequeñas (como los aminoácidos) en moléculas mayores (proteínas), y otra fase, llamada catabolismo o fase destructiva, en la que moléculas mayores (glucógeno) se transforman en otras más pequeñas (ácido pirúvico) liberando energía en el proceso. Estos dos procesos son conjugados y cada uno depende del otro. Estas reacciones bioquímicas están organizadas de forma que se siga siempre una determinada ruta metabólica, de tal forma que un sustrato determinado es transformado en un producto concreto, y éste a su vez es el sustrato para crear otro producto. Dentro de esta vía existen unas proteínas llamadas enzimas que posibilitan estas reacciones, comportándose como factores reguladores de estas rutas metabólicas. El siguiente cuestionario tiene la finalidad de profundizar en algunos conceptos importantes dentro de la fisiopatología del metabolismo.

Delivery of antigen to nasal-associated lymphoid tissue microfold cells through secretory IgA targeting local dendritic cells confers protective immunity.

BACKGROUND: Transmission of mucosal pathogens relies on their ability to bind to the surfaces of epithelial cells, to cross this thin barrier, and to gain access to target cells and tissues, leading to systemic infection. This implies that pathogen-specific immunity at mucosal sites is critical for the control of infectious agents using these routes to enter the body. Although mucosal delivery would ensure the best onset of protective immunity, most of the candidate vaccines are administered through the parenteral route. OBJECTIVE: The present study evaluates the feasibility of delivering the chemically bound p24gag (referred to as p24 in the text) HIV antigen through secretory IgA (SIgA) in nasal mucosae in mice. RESULTS: We show that SIgA interacts specifically with mucosal microfold cells present in the nasal-associated lymphoid tissue. p24-SIgA complexes are quickly taken up in the nasal cavity and selectively engulfed by mucosal dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin-positive dendritic cells. Nasal immunization with p24-SIgA elicits both a strong humoral and cellular immune response against p24 at the systemic and mucosal levels. This ensures effective protection against intranasal challenge with recombinant vaccinia virus encoding p24. CONCLUSION: This study represents the first example that underscores the remarkable potential of SIgA to serve as a carrier for a protein antigen in a mucosal vaccine approach targeting the nasal environment.

Metodología para el Desarrollo Automatizado de Aplicaciones Seguras Basadas en Agentes Móviles

En este trabajo presentamos una metodología para el diseño de aplicaciones basadas en agentes móviles seguros, y unas herramientas para aplicar esta metodología. Los recientes avances en el campo de la seguridad de los agentes móviles pueden desbloquear el uso de esta tecnología, pero falta aún superar la complejidad que suponen al programador de las aplicaciones. La metodología propuesta, y el uso de nuestras herramientas, permiten agilizar el proceso de diseño de arquitecturas criptográficas, y de los agentes que las utilizan. La implementación presentada es muy flexible, y permite utilizarse con cualquier plataforma de agentes que siga los estándares de IEEE-FIPA y que permita los esquemas de protección conducidos por agente.

Duração e porcentagem de molhamento foliar determinados pelo espaçamento entrelinhas, e influência sobre a ferrugem asiática da soja

A ferrugem asiática da soja causada pelo fungo Phakopsora pachyrhizi, é a doença de maior virulência e rápida disseminação da cultura no Brasil. Para que haja a infecção nas plantas são necessárias condições ambientais favoráveis, principalmente molhamento foliar quantificado pela duração do período de molhamento (DPM). Alterações no espaçamento entrelinhas de cultivo podem modificar a DPM. Este trabalho teve como objetivo verificar o efeito da utilização de dois espaçamentos entrelinhas sobre a duração e porcentagem do molhamento foliar, e a influência sobre a infecção inicial e desenvolvimento da doença. O experimento foi conduzido nas safras 2011/2012 e 2012/2013 na fazenda da Universidade Estadual de Londrina (23º34' S / 51º21' W), onde foram instalados coletores de esporo SIGA para identificar os uredósporos da P. pachyrhizi. Para quantificar a DPM e sua porcentagem foram utilizadas Árvores Eletrônicas de Molhamento, com sensores em três alturas (0,9m; 0,6m; 0,3m), nos espaçamentos entrelinhas de 0,45 e 0,8 m. Nas safras avaliadas foram detectados uredósporos da P. pachyrhizi antes do aparecimento dos primeiros sintomas. Foram necessárias cerca de 6 h de DPM acima 50% para que houvesse a infecção e manifestação dos sintomas de ferrugem. Foram detectadas 2 h a mais de DPM acima de 50% no terço médio (0,6 m) no espaçamento de 0,45 m em relação ao de 0,8 m em ambas as safras, no entanto não houve diferenciação na severidade entre os espaçamentos. A maior quantidade de uredósporos detectados e o volume de chuva podem explicar a maior severidade final de ferrugem na safra 2012/2013.