The NF-κB signaling protein Bcl10 regulates actin dynamics by controlling AP1 and OCRL-bearing vesicles.

The protein Bcl10 contributes to adaptive and innate immunity through the assembly of a signaling complex that plays a key role in antigen receptor and FcR-induced NF-κB activation. Here we demonstrate that Bcl10 has an NF-κB-independent role in actin and membrane remodeling downstream of FcR in human macrophages. Depletion of Bcl10 impaired Rac1 and PI3K activation and led to an abortive phagocytic cup rich in PI(4,5)P(2), Cdc42, and F-actin, which could be rescued with low doses of F-actin depolymerizing drugs. Unexpectedly, we found Bcl10 in a complex with the clathrin adaptors AP1 and EpsinR. In particular, Bcl10 was required to locally deliver the vesicular OCRL phosphatase that regulates PI(4,5)P(2) and F-actin turnover, both crucial for the completion of phagosome closure. Thus, we identify Bcl10 as an early coordinator of NF-κB-mediated immune response with endosomal trafficking and signaling to F-actin remodeling.

Crystallographic fatigue crack growth in nimonic AP1

Recently a controversy has developed over whether crystallographic crack growth near threshold in nickel-base superalloys occurs along right brace 111 left brace slip planes or right brace 100 left brace planes at room temperature. In this work crack propagation is shown to occur on both right brace 100 left brace and right brace 111 left brace planes. The most common facet plane is right brace 111 left brace and this is the only orientation observed at the lowest stress intensities, but at higher stress intensities occasional right brace 100 left brace facets are also produced. This behavior is compared with similar results in aluminum alloys.

Correlation between c-Jun and human papillomavirus in oral premalignant and malignant lesions

c-Jun, one of the components of the transcription factor activating protein-1 (AP-1), is suggested as a factor in malignant progression of oral lesions. c-Jun and other AP-1 components relationships with human papillomavirus (HPV) infection have been investigated, but not yet focusing on oral carcinogenesis. The aim of this study was to verify whether c-Jun immunohistochemical expression is related to HPV DNA detection in oral premalignant and malignant lesions. Fifty cases diagnosed as oral leukoplakias, with different degrees of epithelial dysplasia, and as oral squamous cell carcinomas (OSCC) were submitted to immunohistochemistry to detect c-Jun and to in situ hybridization with signal amplification to assess HPV DNA. It was verified that c-Jun nuclear expression increased according to the degree of dysplasia within the lesion, with the greatest expression in OSCC. The same did not happen concerning HPV infection - a discrete proportional relation was observed in indexes found in leukoplakia with no dysplasia, leukoplakia with dysplasia and OSCC, but statistically insignificant. When separating the group of leukoplakia by degrees of dysplasia, this relation of proportion was not observed. Nevertheless, the overall prevalence of HPV infection was 24% and the high-risk HPV types were the most frequently identified, which does not allow excluding HPV as a risk factor in oral carcinogenesis. When relating c-Jun expression and HPV infection, no statistically significant relationship is observed. Results suggest then that malignant progression mediated by c-Jun is independent of the presence of HPV in oral carcinogenesis. (C) 2007 Elsevier Ltd. All rights reserved.

Site-directed mutagenesis of the ATM promoter: Consequences for response to proliferation and ionizing radiation

Although ATM, the protein defective in ataxia-telangiectasia (A-T), is activated primarily by radiation, there is also evidence that expression of the protein can be regulated by both radiation and growth factors. Computer analysis of the ATM promoter proximal 700-bp sequence reveals a number of potentially important cis-regulatory sequences. Using nucleotide substitutions to delete putative functional elements in the promoter of ATM, we examined the importance of some of these sites for both the basal and the radiation-induced activity of the promoter. In lymphoblastoid cells, most of the mutations in transcription factor consensus sequences [Sp1(1), Sp1(2), Cre, Ets, Xre, gammaIre(2), a modified AP1 site (Fse), and GCF] reduced basal activity to various extents, whereas others [gammaIre(1), NF1, Myb] left basal activity unaffected. In human skin fibroblasts, results were generally the same, but the basal activity varied up to 8-fold in these and other cell lines. Radiation activated the promoter approximately 2.5-fold in serum-starved lymphoblastoid cells, reaching a maximum by 3 hr, and all mutated elements equally blocked this activation. Reduction in Sp1 and AP1 DNA binding activity by serum starvation was rapidly reversed by exposure of cells to radiation. This reduction was not evident in A-T cells, and the response to radiation was less marked. Data provided for interaction between ATM and Sp1 by protein binding and co-immunoprecipitation could explain the altered regulation of Sp1 in A-T cells. The data described here provide additional evidence that basal and radiation-induced regulation of the ATM promoter is under multifactorial control. (C) 2003 Wiley-Liss, Inc.

Study of the retinoic acid regulated genes in the rodent hypothalamus controlling growth and appetite and the potential for the epigenetic control

Dissertação de mestrado em Bioquímica (área de especialização em Biomedicina)

A trecut badità dealul (Il a passé la colline) : doïna de Transylvanie ; Trage i Radul di cimpoi (Chante, Radoulé, de la cornemuse) : doïna de Moldavie / Michel Vulpesco, baryton ; accompagnement de piano

Langue roumaine

Genesis of textural contrasts in subsurface soil horizons in the Northern Pantanal-Brazil

The Pantanal region can be characterized as a quaternary floodplain with predominant sedimentation in the form of alluvial fans. In the geomorphologic and sedimentary evolution, the avulsion process is inherent to this depositional system and its dynamics, together with surface water floods, influence soil sedimentation on this plain. The knowledge and differentiation of these two events can contribute to a better understanding of the variability of soil properties and distribution under the influence of these sedimentation processes. Therefore, this study investigated the genesis of soils in the Northern Pantanal with textural contrasts in deeper horizons and their relationship with the depositional system dynamics. We analyzed four soil profiles in the region of Barão de Melgaço, Mato Grosso State, Brazil (RPPN SESC Pantanal). Two profiles were sampled near the Rio Cuiabá (AP1 and AP4) and two near the Rio São Lourenço (AP10 and AP11). In AP11, the horizons contrast in particle size between the profile basis and the surface. In AP1, AP4 and AP10, the horizons overlaying the sand layer have similar particle size properties, mainly in terms of sand distribution. In the first case, floods (surface water) seem to have originated the horizons and layers with contrasting texture. In the second case, avulsion is the most pronounced process. Therefore, the two modes can form soils with contrasting texture that are discriminable by soil morphology, based on the distinct features associated to the specific sedimentation processes.

Molecular insights into T cell activation : study of the TRAF, Bcl10 and Carma 1 proteins

Abstract : Activation of naïve T lymphocytes is essential for the onset of an adaptive immune response against a pathogenic threat. T lymphocytes are activated through the engagement of their highly specific cell surface antigen-receptor (TCR), together with co-stimulatory receptors, by activated antigen-presenting cells that display antigenic peptide fragments from the pathogen that they have detected. Dissection of the mechanisms that modulate TCR- and co-stimulation- induced signals is therefore crucial for the understanding of the molelcular basis of adaptive immune responses. Following antigen-receptor triggering, the Carma1, Bcl10 and Malt1 (CBM) proteins assemble into an oligomeric complex, which is essential for activation of the NF-κB and JNK signaling pathways in lymphocytes. In this work, by using human epithelial and lymphocytic cell lines, we identified the TNF-receptor-associated factor (TRAF) proteins TRAF3 and TRAF7 as new binding partners of Bcl10 and Carma1, respectively. We could show that TRAF3 is required for the proper transcriptional upregulation of IL-2 in activated T cells, and that endogenous TRAF3 is recruited to Bcl10 following TCR engagement. Although the mechanisms used by TRAF3 to modulate the transcriptional activation of the IL-2 promoter are not elucidated, the stimulus-dependent association ofTRAF3 with its direct binding partner Bcl10 suggests that TRAF3 is regulating Bcl10 function in TCR-activated lymphocytes. We also demonstrated that TRAF7 acts as a negative regulator of Carma1-induced NFκB-and AP1-dependent transcription by overexpression in 293T cells. These data suggest that TRAF7 could contribute to the negative regulation of TCR-dependent Carma1 functions. Finally, we showed that Carma1 is processed upon antigen-receptor triggering in B and T cell lines, as well as in primary human CTLs, and that this processing is dependent on the proteolytic activity of Malt1. Collectively, this work contributes to describe new proteins and regulatory mechanisms that modulate CBM-dependent functions in activated lymphocytes. Furthermore, it uncovers new tracks that could lead to a better molecular understanding of the complex interplay between the activatory and inhibitory regulators associated with the CBM complex. Résumé : L'activation des lymphocytes T naifs est une étape essentielle à la mise en place d'une réponse immunitaire adaptative pour combattre une infection. Après la détection d'un pathogène, les cellules présentatrices d'antigènes exposent à leur surface des fragments peptidiques provenant du pathogène, qui activent le récepteur à antigène (TCR) spécifique des lymphocytes T, ainsi que des molécules co-stimulatrices qui contribuent à l'activation complète des lymphocytes T. La caractérisation des mécanismes qui modulent les cascades de signaux émanant du TCR et des récepteurs de co-stimulation est essentielle à la compréhension du fonctionnement moléculaire de la réponse immunitaire adaptative. La ligation du TCR induit la formation d'un complexe oligomérique comprenant les protéines Carma1, Bcl10 et Malt1, qui est essentiel à l'activation des voies de signalisation cellulaires NF-κB et JNK induisant l'activation complète des lymphorctes T. Dans cette étude, à l'aide de lignées de cellules humaines épithéliales et lymphocytaires, nous avons identifié que deux protéines de la famille des TRAF (Tumor Necrosis Factor Receptor-Associated Factor), TRAF3 et TRAF7, s'associent à Bc110 et à Carma1, respectivement. Les TRAFs sont d'importants régulateurs des voies de signalisation dans les cellules du système immunitaire inné et adaptatif. Nous avons démontré que TRAF3 était important pour permettre la transcription de l'interleukine-2 (IL-2) dans les lymphocytes T activés, et que TRAF3 s'associait à Bc110 à la suite de la stimulation du TCR Les mécanismes que TRAF3 utilise pour moduler l'activation du promoteur de l'IL-2 ne sont pas connus, mais l'association de TRAF3 à Bc110 suite à la stimulation du TCR suggère que TRAF3 régule la fonction de Bc110. Nous avons également identifié TRAF7 comme un nouveau régulateur négatif des voies NF-κB et JNK induites par surexpression de la protéine Carma1. Nos données suggèrent que TRAF7 pourrait également contribuer à la régulation négative de la fonction de Carma1 dans les lymphocytes activés. Enfin, nous avons découvert que Carma1 était clivé suite à la stimulation du TCR, et que ce clivage dépendait de l'activité protéolytique de Malt1. Cette étude contribue ainsi à la description de nouvelles protéines et de nouveaux mécanismes qui modulent l'activité du complexe CBM dans les lymphocytes activés, et ouvre la voie à la caractérisation moléculaire de ces nouveaux mécanismes importants pour la régulation de la réponse immunitaire adaptative.

The Oncogene ATF3 Is Potentiated by Cyclosporine A and Ultraviolet Light A.

Cutaneous squamous cell carcinoma (SCC) represents the most important cutaneous complication following organ transplantation. It develops mostly on sun-exposed areas. A recent study showed the role of activating transcription factor 3 (ATF3) in SCC development following treatment with calcineurin inhibitors. It has been reported that ATF3, which may act as an oncogene, is under negative calcineurin/nuclear factor of activated T cells (NFAT) control and is upregulated by calcineurin inhibitors. Still, these findings do not fully explain the preferential appearance of SCC on chronically sun-damaged skin. We analyzed the influence of UV radiation on ATF3 expression and its potential role in SCC development. We found that ATF3 is a specifically induced AP1 member in SCC of transplanted patients. Its expression was strongly potentiated by combination of cyclosporine A and UVA treatment. UVA induced ATF3 expression through reactive oxygen species-mediated nuclear factor erythroid 2-related factor 2 (NRF2) activation independently of calcineurin/NFAT inhibition. Activated NRF2 directly binds to ATF3 promoter, thus inducing its expression. These results demonstrate two mechanisms that independently induce and, when combined together, potentiate the expression of ATF3, which may then force SCC development. Taking into account the previously defined role of ATF3 in the SCC development, these findings may provide an explanation and a mechanism for the frequently observed burden on SCCs on sun-exposed areas of the skin in organ transplant recipients treated by calcineurin inhibitors.

Produtividade e qualidade de frutos de progênies de maracujazeiro-azedo cultivados no Distrito Federal

Este trabalho teve como objetivo avaliar a produtividade e a qualidade de frutos de progênies de maracujazeiro-azedo cultivados no Distrito Federal. Foi conduzido na Fazenda Água Limpa-UnB, utilizando-se de delineamento de blocos casualizados, com 14 tratamentos (progênies), 04 repetições, sendo a parcela útil de oito plantas. As progênies avaliadas foram: Vermelhão Ingaí, EC-RAM, AR01, AR02, mar20#36, mar20#46, AP1, FP01, FB200, RC3, GA2-A1*AG, mar20#03, mar20#23 e mar20#09. As características analisadas em 75 colheitas foram: produtividade (kg/ha), massa média de frutos (g), número de frutos por hectare, coloração da casca e classificação quanto ao tamanho. Houve maior rendimento em Vermelhão Ingaí, EC-RAM e AR01. O Vermelhão Ingaí produziu a maior quantidade de frutos por hectare, estatisticamente superior aos demais. O EC-RAM obteve o maior número de frutos rosa e roxo. Todas as progênies apresentaram maior produção de frutos de cor amarela. O peso médio total variou de 83 g em mar20#36 e 128 g em EC-RAM. A maior produtividade de frutos para fins industriais foi verificado em Vermelhão Ingaí com frutos de primeira e mar20#46 com frutos 1B. Já para consumo in natura foi observado em AR02 com frutos 1A e 2 A e em AR01 com frutos 3 A.

Caractérisation fonctionnelle du gène AP1S1 mutant associé au syndrome de MEDNIK

Dans les cellules eucaryotes, le trafic intracellulaire de nombreuses protéines est assuré par des vésicules de transport tapissées de clathrine. Les complexes adaptateurs de clathrine (AP) sont responsables de l’assemblage de ces vésicules et de la sélection des protéines qui seront transportées. Nous avons étudié cinq familles atteintes du syndrome neurocutané MEDNIK qui est caractérisé par un retard mental, une entéropathie, une surdité, une neuropathie périphérique, de l’icthyose et de la kératodermie. Tous les cas connus de cette maladie à transmission autosomique récessive sont originaires de la région de Kamouraska, dans la province de Québec. Par séquençage direct des gènes candidats, nous avons identifié une mutation impliquant le site accepteur de l’épissage de l’intron 2 du gène codant pour la sous-unité σ1 du complexe AP1 (AP1S1). Cette mutation fondatrice a été retrouvée chez tous les individus atteints du syndrome MEDNIK et altère l’épissage normal du gène, menant à un codon stop prématuré. Afin de valider l’effet pathogène de la mutation, nous avons bloqué la traduction de cette protéine chez le poisson zébré en injectant une séquence d’oligonucléotides antisenses spécifique à AP1S1. À 48 heures après la fertilisation, les larves knock down pour AP1S1 montrent une réduction de la pigmentation, une désorganisation de la structure de l’épiderme et une perturbation du développement moteur. Alors que la surexpression de l’AP1S1 humain dans ce modèle a permis la récupération du phénotype normal, l’expression de l’AP1S1 mutant fut sans effet sur les phénotypes moteurs et cutanés des larves knock down. Les résultats obtenus montrent que la mutation du AP1S1 responsable du syndrome de MEDNIK est associée à une perte de fonction et que la sous-unité σ1 du complexe AP1 joue un rôle crucial dans l’organisation de l’épiderme et le développement de la moelle épinière.

Efecto de la hipoxia-reoxigenación y las radiaciones ionizantes en la captación de glucosa en líneas tumorales de seno y colon cocultivadas con células endoteliales.

La captación de glucosa y su conversión en lactato juega un papel fundamental en el metabolismo tumoral, independientemente de la concentración de oxígeno presente en el tejido (efecto Warburg). Sin embrago, dicha captación varía de un tipo tumoral a otro, y dentro del mismo tumor, situación que podría depender de las características microambientales tumorales (fluctuaciones de oxígeno, presencia de otros tipos celulares) y de factores estresores asociados a los tratamientos. Se estudió el efecto de la hipoxia-reoxigenación (HR) y las radiaciones ionizantes (RI) sobre la captación de glucosa, en cultivos de líneas tumorales MCF-7 y HT-29, cultivadas de forma aislada o en cocultivo con la línea celular EAhy296. Se encontró que la captación de glucosa en HR es diferente para lo descrito en condiciones de hipoxia permanente y que es modificada en el cocultivo. Se identificaron poblaciones celulares dentro de la misma línea celular, de alta y baja captación de glucosa, lo que implicaría una simbiosis metabólica de la célula como respuesta adaptativa a las condiciones tumorales. Se evaluó la expresión de NRF2 y la translocación nuclear de NRF2 y HIF1a, como vías de respuesta a estrés celular e hipoxia. La translocación nuclear de las proteínas evaluadas explicaría el comportamiento metabólico de las células tumorales de seno, pero no de colon, por lo cual deben existir otras vías metabólicas implicadas. Las diferencias en el comportamiento de las células tumorales en HR en relación con hipoxia permitirá realizar planeaciones dosimétricas más dinámicas, que reevalúen las condiciones de oxigenación tumoral constantemente.

Condiciones para preservar e valor moral del consentimiento informado en medicina

El consentimiento informado (CI) en medicina corre el riesgo de quedar reducido a un formalismo legal sin valor moral; esto puede suceder por dos cosas: 1) como consecuencia de que el paciente no comprenda con suficiencia el concepto de autonomía y 2) debido a la disminución de la autonomía práctica del paciente por efecto de la autoridad del personal médico o de las instituciones de salud. Este artículo tiene como fin mostrar bajo qué condiciones se puede evitar la reducción del CI a un formalismo legal desprovisto de valor moral. Para servir a este propósito, se considera el concepto de autonomía relacional como eje articulador que pone en relación al paciente con su comunidad moral y con el médico, lo que impulsa al personal de la salud a proporcionar las condiciones para tal fin. Así, se afirma que la valía moral del CI puede conservarse, primero, a partir de una concepción relacional de la autonomía y, segundo, mediante la aplicación, por parte del médico y de la institución, de un protocolo que disminuya la posibilidad de choque entre la autonomía y la autoridad.

Resultados de la terapia antiangiogenica con Ranibizumab en retinopatia del prematuro: revisión sistemática.

Introducción: La retinopatía del prematuro (ROP) se presenta hasta en el 73% de los neonatos pretérmino por debajo de la semana 27, los agentes antiangiogénicos se presentan como una herramienta para su tratamiento con resultados prometedores. En el presente estudio se pretendió evaluar los resultados de la terapia antiangiogénica con ranibizumab en pacientes con retinopatía del prematuro según la evidencia descrita hasta la fecha. Metodología: se realizó una revisión sistemática de literatura con componente meta-analítico de los datos sociodemográficos y clínicos, incluyendo todos los artículos sobre el tema sin límite de fecha de publicación. Se usó una estrategia de búsqueda en diferentes bases de datos, todos los estudios relacionados con el pronóstico de la enfermedad fueron elegibles. Resultados: Se encontraron un total de 13 artículos que cumplieron criterios de elegibilidad para su inclusión, que incluyeron 75 pacientes en total (133 ojos evaluados). La edad promedio al nacimiento fue 23.6 semanas, la edad al momento de aplicación de tratamiento fue 36.3 semanas. 11/13 artículos reportaron que el tratamiento con ranibizumab fue satisfactorio en términos de resolución completa de la enfermedad sin efectos secundarios (64 pacientes) Discusión: Los resultados de la terapia con ranibizumab para retinopatía del prematuro según la evidencia hasta la fecha permiten recomendar el uso de terapia antiangiogénica con el fin de mejorar la salud visual a mediano y largo plazo y por ende disminución en la prevalencia de ceguera por esta causa.