The histone-interacting domain of nuclear factor I activates simian virus 40 DNA replication in vivo.

Efficient initiation of SV40 DNA replication requires transcription factors that bind auxiliary sequences flanking the minimally required origin. To evaluate the possibility that transcription factors may activate SV40 replication by acting on the chromatin structure of the origin, we used an in vivo replication system in which we targeted GAL4 fusion proteins to the minimally required origin. We found that the proline-rich transcriptional activation domain of nuclear factor I (NF-I), which has been previously shown to interact with histone H3, specifically activates replication. Evaluation of a series of deletion and point mutants of NF-I indicates that the H3-binding domain and the replication activity coincide perfectly. Assays with other transcription factors, such as Sp1, confirmed the correlation between the interaction with H3 and the activation of replication. These findings imply that transcription factors such as NF-I can activate SV40 replication via direct interaction with chromatin components, thereby contributing to the relief of nucleosomal repression at the SV40 origin.

Proteasome-assisted identification of a SSX-2-derived epitope recognized by tumor-reactive CTL infiltrating metastatic melanoma.

The tumor Ag SSX-2 (HOM-MEL-40) was found by serological identification of Ags by recombinant expression cloning and was shown to be a cancer/testis Ag expressed in a wide variety of tumors. It may therefore represent a source of CD8(+) T cell epitopes useful for specific immunotherapy of cancer. To identify potential SSX-2-derived epitopes that can be recognized by CD8(+) T cells, we used an approach that combined: 1) the in vitro proteasomal digestion of precursor peptides overlapping the complete SSX-2 sequence; 2) the prediction of SSX-2-derived peptides with an appropriate HLA-A2 binding score; and 3) the analysis of a tumor-infiltrated lymph node cell population from an HLA-A2(+) melanoma patient with detectable anti-SSX-2 serum Abs. This strategy allowed us to identify peptide SSX-2(41-49) as an HLA-A2-restricted epitope. SSX2(41-49)-specific CD8(+) T cells were readily detectable in the tumor-infiltrated lymph node population by multimer staining, and CTL clones isolated by multimer-guided cell sorting were able to lyse HLA-A2(+) tumor cells expressing SSX-2.

Identification of in vitro phosphorylation sites in the growth cone protein SCG10. Effect Of phosphorylation site mutants on microtubule-destabilizing activity.

SCG10 is a neuron-specific, membrane-associated protein that is highly concentrated in growth cones of developing neurons. Previous studies have suggested that it is a regulator of microtubule dynamics and that it may influence microtubule polymerization in growth cones. Here, we demonstrate that in vivo, SCG10 exists in both phosphorylated and unphosphorylated forms. By two-dimensional gel electrophoresis, two phosphoisoforms were detected in neonatal rat brain. Using in vitro phosphorylated recombinant protein, four phosphorylation sites were identified in the SCG10 sequence. Ser-50 and Ser-97 were the target sites for protein kinase A, Ser-62 and Ser-73 for mitogen-activated protein kinase and Ser-73 for cyclin-dependent kinase. We also show that overexpression of SCG10 induces a disruption of the microtubule network in COS-7 cells. By expressing different phosphorylation site mutants, we have dissected the roles of the individual phosphorylation sites in regulating its microtubule-destabilizing activity. We show that nonphosphorylatable mutants have increased activity, whereas mutants in which phosphorylation is mimicked by serine-to-aspartate substitutions have decreased activity. These data suggest that the microtubule-destabilizing activity of SCG10 is regulated by phosphorylation, and that SCG10 may link signal transduction of growth or guidance cues involving serine/threonine protein kinases to alterations of microtubule dynamics in the growth cone.

Nuclear translocation and DNA recognition signals colocalized within the bZIP domain of cyclic adenosine 3',5'-monophosphate response element-binding protein CREB.

CREB is a cAMP-responsive nuclear DNA-binding protein that binds to cAMP response elements and stimulates gene transcription upon activation of the cAMP signalling pathway. The protein consists of an amino-terminal transcriptional transactivation domain and a carboxyl-terminal DNA-binding domain (bZIP domain) comprised of a basic region and a leucine zipper involved in DNA recognition and dimerization, respectively. Recently, we discovered a testis-specific transcript of CREB that contains an alternatively spliced exon encoding multiple stop codons. CREB encoded by this transcript is a truncated protein lacking the bZIP domain. We postulated that the antigen detected by CREB antiserum in the cytoplasm of germinal cells is the truncated CREB that must also lack its nuclear translocation signal (NTS). To test this hypothesis we prepared multiple expression plasmids encoding carboxyl-terminal deletions of CREB and transiently expressed them in COS-1 cells. By Western immunoblot analysis as well as immunocytochemistry of transfected cells, we show that CREB proteins truncated to amino acid 286 or shorter are sequestered in the cytoplasm, whereas a CREB of 295 amino acids is translocated into the nucleus. Chimeric CREBs containing a heterologous NTS fused to the first 248 or 261 amino acids of CREB are able to drive the translocation of the protein into the nucleus. Thus, the nine amino acids in the basic region involved in DNA recognition between positions 287 and 295 (RRKKKEYVK) of CREB contain the NTS. Further, mutation of the lysine at position 290 in CREB to an asparagine diminishes nuclear translocation of the protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Simulación de los cambios de carbono orgánico del suelo en sistema de cultivo con higuerilla por el modelo RothC

El objetivo del presente trabajo fue la simulación de los cambios del carbono orgánico del suelo (COS), por el modelo RothC, en razón del cambio de uso de suelo del sistema tradicional - asociación maíz-calabaza (TMC) - a sistemas con higuerilla (Ricinus communis): multiestratos (MUL), callejones (CALL) y monocultivo de higuerilla (HIG). Las simulaciones del COS se hicieron para las profundidades de suelo 0-20 y 0-40 cm, para el periodo de 1980-2040, considerándose como línea base al sistema TMC. Las tasas de cambio de COS estimadas con el RothC, en ambas profundidades de suelo, estuvieron en 0,5-1,2, 0,4-0,8, 0,3-0,5 y 0,04-0,1 Mg ha-1 de C por año en los sistemas HIG, MUL, CALL y TMC, respectivamente, y fueron consistentes con las reportadas en la literatura. El desempeño del RothC tuvo 89% de eficiencia (EF), con R²=0,9, lo que muestra que este modelo puede usarse con información temporal del COS escasa, información de la historia de uso de suelo y mediciones de la entrada de residuos vegetales aéreos y subterráneos en el suelo.

Selective cooperation between fatty acid binding proteins and peroxisome proliferator-activated receptors in regulating transcription.

Lipophilic compounds such as retinoic acid and long-chain fatty acids regulate gene transcription by activating nuclear receptors such as retinoic acid receptors (RARs) and peroxisome proliferator-activated receptors (PPARs). These compounds also bind in cells to members of the family of intracellular lipid binding proteins, which includes cellular retinoic acid-binding proteins (CRABPs) and fatty acid binding proteins (FABPs). We previously reported that CRABP-II enhances the transcriptional activity of RAR by directly targeting retinoic acid to the receptor. Here, potential functional cooperation between FABPs and PPARs in regulating the transcriptional activities of their common ligands was investigated. We show that adipocyte FABP and keratinocyte FABP (A-FABP and K-FABP, respectively) selectively enhance the activities of PPARgamma and PPARbeta, respectively, and that these FABPs massively relocate to the nucleus in response to selective ligands for the PPAR isotype which they activate. We show further that A-FABP and K-FABP interact directly with PPARgamma and PPARbeta and that they do so in a receptor- and ligand-selective manner. Finally, the data demonstrate that the presence of high levels of K-FABP in keratinocytes is essential for PPARbeta-mediated induction of differentiation of these cells. Taken together, the data establish that A-FABP and K-FABP govern the transcriptional activities of their ligands by targeting them to cognate PPARs in the nucleus, thereby enabling PPARs to exert their biological functions.

Going with the consumer towards the social Web environment : a review of extant knowledge

The social Web environment cannot be explained without taking into account the use made by consumers of the web technologies in the field of e-commerce. Starting with a systematic background of the growing body of research on marketing and consumer behaviour on the Web; this article presents a critical review of the main contributions on this theme, integrating also the research field of consumer behaviour in the social Web environment. It concludes with some managerial guidelines for successful use of the social Web in the commercial activities and in the establishment of fruitful relationships between the consumers and the brand.

The cAMP response element binding protein-2 (CREB-2) can interact with the C/EBP-homologous protein (CHOP).

cAMP response element binding protein-2 (CREB-2) is a basic leucine zipper (bZIP) factor that was originally described as a repressor of CRE-dependent transcription but that can also act as a transcriptional activator. Moreover, CREB-2 is able to function in association with the viral Tax protein as an activator of the human T-cell leukemia virus type I (HTLV-I) promoter. Here we show that CREB-2 is able to interact with C/EBP-homologous protein (CHOP), a bZIP transcription factor known to inhibit CAAT/enhancer-dependent transcription. Cotransfection of CHOP with CREB-2 results in decreased activation driven by the cellular CRE motif or the HTLV-I proximal Tax-responsive element, confirming that CREB-2 and CHOP can interact with each other in vivo.

Experimental Results and Evaluation of SmartBox Stimulation Device in a P2P E-learning System

In this paper, we present the experimental results and evaluation of the SmartBox stimulation device in P2P e-learning system which is based on JXTA-Overlay. We also show the design and implementation of the SmartBox environment that is used for stimulating the learners motivation to increase the learning efficiency. The SmartBox is integrated with our P2P system as a useful tool for monitoring and controlling learners¿ activity. We found by experimental results that the SmartBox is an effective way to increase the learner¿s concentration. We also investigated the relation between learner¿s body movement, concentration, and amount of study. From the experimental results, we conclude that the use of SmartBox is an effective way to stimulate the learners in order to continue studying while maintaining the concentration.

Donne e lavoro. Prospettive per una storia delle montagne europeee XVIII-XX secc.

Si ritiene comunemente che le spinte emancipatrici femminili si siano imposte con maggior vigore nei luoghi contraddistinti da mercati lavorativi più sviluppati e da maggiori opportunità di impiego retribuito, mentre nei contesti periferici, caratterizzati da marginalità economica, queste spinte sarebbero state più flebili. Il ritardo della modernizzazione di molte regioni di montagna sarebbe quindi responsabile del perdurare di una realtà femminile segnata da dipendenza e "arretratezza" sociale e culturale. Il volume si propone di analizzare in modo critico il nesso tra marginalità (economica, culturale, geografica) e discriminazione femminile, ampliando le prospettive di genere e contribuendo così al dibattito in corso da diversi anni sulle società di montagna del passato e sul loro divenire all'interno della realtà europea. I contributi abbracciano diversi contesti montani europei e prendono in esame alcuni nodi tematici quali i profili dei ruoli femminili, il loro inserimento nel mercato lavorativo e le rappresentazioni che scaturiscono dalle loro attività, delineando molteplici prospettive per lo studio del binomio genere-montagna.

Concurs per a una Llar d'Infants

El projecte defineix un carrer interior tangent a l’edifici del pavelló per on es produeix l’entrada i per on s’atravessa visualment l’edifici, de manera que la façana del pavelló te continuïtat cap a l’interior de la llar d’infants. Perpendicularment es disposa l’espai del vestíbul que articula els espais de circulació i dona sortida i visuals al pati de jocs. Les zones de pas tenen una alçada inferior a la de les altres àrees per lògica d’us i per donar claredat volumètrica a l’edifici. L’entrada es produeix per una rampa, sota un voladís que a l’hora recull l’escletxa oberta a la façana del pavelló per il•luminar l’àrea administrativa. L’altre extrem del carrer és una sortida d’emergència. En una visió general s’observa que el pla de façana del pavelló fa de suport a dos volums que entren en relació: la rampa d’accés al pavelló i la coberta del carrer interior de la llar d’infants. Pel que fa a les alçades, el conjunt manté el criteri de donar major alçada als espais principals (pista i aules) respecte els d’accés. Pel que fa a la forma, el cos d’aules adopta la coberta plana per entrar en relació amb l’horitzontalitat de la visera de la façana principal del pavelló i no la forma corba perquè l’escala del cos d’aules no admet aquesta solució. S’enfatitzaria al màxim la percepció de conjunt si les cobertes de les aules i del pavelló fossin igualment inclinades. Les dependències s’organitzen en dues àrees: L’àrea administrativa es col•loca sota el forjat que defineix el projecte del pavelló. Atès que l’alçada lliure interior és de 3.00 m., i per tal de poder garantir il•luminació i ventil•lació natural a aquestes peces, es proposa baixar la cota de tota la llar d’infants 80 cm. respecte la cota del carrer. Consta d'una sala de reunions propera al vestíbul, un despatx per la direcció del centre uns vestidors amb bany per al personal i una sala d'usos múltiples. L’àrea d’activitats es col•loca separada del pavelló, definint un volum independent, tant en planta com en secció. Consta de 3 aules de 20 alumnes que donen resposta a les necessitats plantejades pel programa. Aquestes s’obren generosament al pati de jocs buscant la llum de nord i per la part superior amb una obertura molt més controlada tenen la llum del sud. Això permet il•luminar i ventil•lar de forma creuada. El tram de carrer interior davant de les aules es converteix en una zona de jocs comuna.

Prematuritat i dol

Quan parlem de la prematuritat, tot ens porta a pensar en els avenços de la tècnica, en els nens tan i tan petits que salvem, però poques vegades ens aturem a pensar en el dol que representa la pèrdua del embaràs o, en el cas del nadó, la pèrdua del cos de la mare, única referència capaç de calmar les ansietats del néixer.

El dibuix de la figura humana com a llenguatge expressiu

El dibuix de la fi gura humana utilitzat com a llenguatge expressiu es pot contemplar a diferents exposicions artístiques com a font d’informació històrica on es relacionen les vivències personals amb la seva representació pictòrica. Les exposicions més representatives d’aquest tema que han tingut lloc a la nostra ciutat en els darrers anys han estat La col·lecció Prinzhorn (Museu d’Art Contemporani de Barcelona, 2001), En guerra (Centre de Cultura Contemporània de Barcelona, 2004) i Hi havia una vegada Txernòbil (Centre de Cultura Contemporània de Barcelona, 2006). En el grup de recerca Salut Activitat Física i Esport (SAFE) de la Facultat de Psicologia Ciències de l’Educació i l’Esport Blanquerna, dins el projecte Dansa Esquema Corporal (DEC) hi ha una línia d’investigació dedicada a avaluar l’impacte de les sessions d’expressió corporal, entesa principalment com a vivència personal, i la seva representació gràfi ca en paper. Els objectius principals de la nostra recerca son examinar si el dibuix de la fi gura humana és una eina efi caç per avaluar el coneixement de l’esquema corporal en població general infantil i en població d’adolescents amb Síndrome de Down amb el test de Goodenough, i avaluar la repercussió de la sessió d’expressió corporal en el desenvolupament emocional amb el test de Koppitz .

Europa als ulls dels Annales (1093-1933)

During this first third of this century, the continent of Europe witnessed far-reaching territorial transformations in the nineteenth century model. To some extent, the setting wich had inspired the epistemological structure of the French regional school was being replaced by one in which -at least to externa1 appearances- the relationship between the physical environment and human activities was much less evident. The adaptation of the theoretical and methodological context to the new Europe seems to take the form of a non-lineal evolution, for in the latter years of the period examined, evolutionary tendencies indicate a return to the more classical model. In this way, the growing interest which the publication had shown in Atlantic Europe and topics of a more economic nature, was truncated. The traditional settings --especially the Mediterranean- came to the fore again and interest in rural societies was renewed. Likewise, the monographic studies which had demonstrated distinct functional objectives in the midtwenties, recovered elements of ecological explanation, and the search for the relationship between environment and society