Ludopoiese e humanescência no educador infantil

This research has as study objective the development of humanescent experience in the process of self-formation of the teachers of the Infantile Educational Unit of the Education Center of the Federal University of Rio Grande do Norte. It has as general objective the description and analysis of how the humanscent experience of life contributes and is manifested in the children educator in his way of being, living and in his pedagogical practice. As specific objectives, the identification of the contributions of the humanescent experiences in the children educator in his way of being, living and in his pedagogical practice; the analysis of the contributions of the humanescent experiences to the children educator from the ludopoetic categories self-esteem, selfworth, self-realization, self-territory and self-connectivity. It is a descriptive study of qualitative nature understood as an existential action research that uses the principles and fundamentals of the etnomethodology and phenomenology. Sixteen teachers six that are effective teachers, six substitute teachers and 4 interns - took part in this investigation. Six humanescent experiences were carried out with them. As main tools we point out: the participative observation, the sand game, the photographic register and the semi-structured interview. The analyses were centered in the categories of the ludopoetic system, described above and referred to in the theoretical fundamentals selected for the study. From the analyses and to the answers to the objectives of the study, we make clear the following considerations: the reports of the teachers reveal that the humanescent experiences have contributed to make them more ludic, sensitive, emotional and lovely people, and this has reflected in their educational practice and has been making sense in their and their students lives. In this way, we make evident that the humanescent experiences have contributed to improve the ludopoesis of the teachers, revealing in the embodiment changes in their way of being, of living together and in their educational practice. In relation to the contributions of the humanescent experiences to the children educator from the ludopoetic categories self-esteem, selfworth, self-realization, self-territory and self-connectivity, we emphasize the following aspects: with the self-esteem, teachers have become more creative, rebuilding their possibilities of intensely experiencing the pleasure of living; in the context of selfterritory, the teachers have provided the realization of the wishes and expresson of oneself while lively interacting with the environment and with the other different territories of the educational process, in a continuous process of demarcation. Concerning the self-connectivity category, we emphasize that the teachers started to become more tuned with themselves, with the environment and with the other teachers because of the general well-being to the success of personal, pedagogical and institutional objectives, promoting a harmonic, lovely and ludic environment. Concerning the self-worth category, the teachers have valued the struggle for a school that is more committed with the search for happiness. From the self-realization, the teachers have shown self-recognition while ludic beings, playing with the beauty in their lives, allowing the flowing in its completeness, the other possibilities of analysis, the educational formation and development. We would like to emphasize that the considerations above are not over after what has been presented because of the richness of experiences revealed in the research

Up-converter nanophosphor Y2O2S:Er,Yb aminofunctionalized containing or not spherical silica conjugated with BSA

This work reports on the study of the nanophosphor. Y2O2S:Er(2%),Yb(1%) obtained from polymeric resin to be evaluated as fluorescent label with Suitable features to conjugate with bio-molecules for bioassay up-converting phosphor technology (UPT) application A conjugation protocol between bovine serum albumin (BSA) and the aminofunctionalized nanophosphor containing or not spherical silica was established UV-vis results indicated an effective conjugation between nanophosphor particles and the protein up-conversion measurements under 980 nm excitation performed for samples before and after aminofunctionalization showed that nanophosphor particles luminescence features keep unchanged in all cases All results suggest that the adapted protocol is feasible to provide a nanoparticle-protein effective conjugation preserving nanophosphor optical features The presence of spherical silica can be considered advantageous to increase conjugation efficiency Therefore. the developed procedure is applicable for future conjugations between the chosen nanophosphor and the streptavidin protein chat takes part in the well known self-recognition system avidin-biotin. (C) 2009 Elsevier B.V All rights reserved.

Blog diários: reflexões sobre a identidade indígena na virtualidade

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Filosofia da história hegeliana: liberdade, razão e o mundo germânico

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Desenvolvimento e aperfeiçoamento de protocolos de conjugação de nanomarcadores luminescentes com sistemas biológicos para aplicação em imunoensaios

Pós-graduação em Química - IBILCE

Síntese e caracterização de híbridos orgânico-inorgânicos luminescentes para aplicação em ensaios biológicos

Pós-graduação em Química - IBILCE

Strukturbildung durch Selbstorganisation von Oligonukleotiden

Die DNA-Doppelhelix ist eine relativ dicke (Ø ≈ 2 nm), kompakte und dadurch auf kurzen Längenskalen relativ steife Verbindung (lp[dsDNA] ≈ 50-60 nm), mit einer klar definierten Struktur, die durch biologische Methoden sehr präzise manipuliert werden kann. Die Auswirkungen der primären Sequenz auf die dreidimensionale Strukturbildung ist gut verstanden und exakt vorhersagbar. Des Weiteren kann DNA an verschiedenen Stellen mit anderen Molekülen verknüpft werden, ohne dass ihre Selbsterkennung gestört wird. Durch die helikale Struktur besteht außerdem ein Zusammenhang zwischen der Lage und der räumlichen Orientierung von eingeführten Modifikationen. Durch moderne Syntheseverfahren lassen sich beliebige Oligonukleotidsequenzen im Bereich bis etwa 150-200 Basen relativ preiswert im Milligrammmaßstab herstellen. Diese Eigenschaften machen die DNA zu einem idealen Kandidaten zur Erzeugung komplexer Strukturen, die durch Selbsterkennung der entsprechenden Sequenzen gebildet werden. In der hier vorgelegten Arbeit wurden einzelsträngige DNA-Abschnitte (ssDNA) als adressierbare Verknüpfungsstellen eingesetzt, um verschiedene molekulare Bausteine zu diskreten nicht periodischen Strukturen zu verbinden. Als Bausteine dienten flexible synthetische Polymerblöcke und semiflexible Doppelstrang-DNA-Abschnitte (dsDNA), die an beiden Enden mit unterschiedlichen Oligonukleotidsequenzen „funktionalisiert“ sind. Die zur Verknüpfung genutzten Oligonukleotidabschnitte wurden so gewählt (n > 20 Basen), dass ihre Hybridisierung zu einer bei Raumtemperatur stabilen Doppelstrangbildung führt. Durch Kombination der Phosphoramiditsynthese von DNA mit einer festkörpergestützten Blockkopplungsreaktion konnte am Beispiel von Polyethylenoxiden ein sehr effektiver Syntheseweg zur Herstellung von ssDNA1-PEO-ssDNA2-Triblockcopolymeren entwickelt werden, der sich problemlos auf andere Polymere übertragen lassen sollte. Die Längen und Basenabfolgen der beiden Oligonukleotidsequenzen können dabei unabhängig voneinander frei gewählt werden. Somit wurden die Voraussetzungen geschaffen, um die Selbsterkennung von Oligonukleotiden durch Kombination verschiedener Triblockcopolymere zur Erzeugung von Multiblockcopolymeren zu nutzen, die mit klassischen Synthesetechniken nicht zugänglich sind. Semiflexible Strukturelemente lassen sich durch die Synthese von Doppelstrangfragmenten mit langen überstehenden Enden (sticky-ends) realisieren. Die klassischen Ansätze der molekularen Genetik zur Erzeugung von sticky-ends sind in diesem Fall nicht praktikabel, da sie zu Einschränkungen im Bezug auf Länge und Sequenz der überhängenden Enden führen. Als Methode der Wahl haben sich zwei verschiedene Varianten der Polymerase Kettenreaktion (PCR) erwiesen, die auf der Verwendung von teilkomplementären Primern beruhen. Die eigentlichen Primersequenzen wurden am 5´-Ende entweder über ein 2´-Desoxyuridin oder über einen kurzen Polyethylenoxid-Spacer (n = 6) mit einer frei wählbaren „sticky-end-Sequenz“ verknüpft. Mit diesen Methoden sind sowohl 3´- als auch 5´-Überhänge zugänglich und die Länge der Doppelstrangabschnitte kann über einen breiten Molmassenbereich sehr exakt eingestellt werden. Durch Kombination derartiger Doppelstrangfragmente mit den biosynthetischen Triblockcopolymeren lassen sich Strukturen erzeugen, die als Modellsysteme zur Untersuchung verschiedener Biomoleküle genutzt werden können, die in Form eines mehrfach gebrochenen Stäbchens vorliegen. Im letzten Abschnitt wurde gezeigt, dass durch geeignete Wahl der überstehenden Enden bzw. durch Hybridisierung der Doppelstrangfragmente mit passenden Oligonukleotiden verzweigte DNA-Strukturen mit Armlängen von einigen hundert Nanometern zugänglich sind. Im Vergleich zu den bisher veröffentlichten Methoden bietet diese Herangehensweise zwei entscheidende Vorteile: Zum einen konnte der Syntheseaufwand auf ein Minimum reduziert werden, zum anderen ist es auf diesem Weg möglich die Längen der einzelnen Arme, unabhängig voneinander, über einen breiten Molmassenbereich zu variieren.

DNA-Polymer-Hybride als molekulare Bausteine

Die DNA hat sich durch die herausstechende Eigenschaft zur Selbstorganisation in den Naturwissenschaften zu einem beliebten Werkzeug entwickelt. In dieser Arbeit wurde die Oligonukleotidselbsterkennung zum Aufbau komplexer Multiblockcopolymere genutzt. Dabei dienten komplementäre einzelsträngige Oligonukleotidsequenzen (ssDNA) als adressierbare Verbindungsstücke zwischen synthetischen Blöcken. Als Bausteine wurden asymmetrische Dreiblockcopolymere der Form DNA1-Polymer-DNA2 aus einer flexiblen Polymereinheit (PEO bzw. PPO) die an beiden Enden mit unterschiedlichen Oligonukleotidsequenzen „funktionalisiert“ ist, verwendet. Diese Bausteine konnten durch die Kombination von Festphasensynthese der Oligonukleotide und Blockkopplung dargestellt werden. Die Oligonukleotidsequenzen wurden so gewählt, dass deren Hybridisierung zu einer bei Raumtemperatur stabilen Verbindung führt. Durch die Verwendung dieser Bausteine erhält man ein modulares System, dass sich durch seine hohe Flexibilität auszeichnet. Aus den dargestellten Dreiblockcopolymeren konnten verschiedene alternierende Multiblockcopolymere aufgebaut werden, wobei die Anzahl der Blöcke (von 11 bis 15) und das PEO / PPO- Verhältnis variiert wurden. Derartige Strukturen sind auf der Grundlage chemischer Synthesen unerreichbar. Die Flexibilität dieses modularen Systems konnte gezeigt werden, indem einzelne Blockbausteine zur Strukturaufklärung einfach ausgetauscht oder weggelassen werden konnten. Durch geeignete Wahl der DNA-Sequenzen konnte zusätzlich das Polymerisationsverhalten dieser Bauelemente untersucht werden. Die Integration längerer kettensteifer DNA-Abschnitte in die Multiblockstrukturen erfolgte durch die Verwendung teilkomplementärer Oligonukleotide. Diese bieten den Vorteil, dass bis zu einer Größe von etwa 150 bp sowohl die Länge als auch die Sequenz der Doppelstrangabschnitte und sticky-ends frei variiert werden können. Die biosynthetischen Dreiblockcopolymere dienten hier als Linkermoleküle zwischen den einzelnen dsDNA-Blöcken. Nach diesem Konzept wurde ein Nonamer als Modellsystem eines mehrfach gebrochenen Stäbchens synthetisiert. Außerdem wurden mit Hilfe der Polymerase Kettenreaktion (PCR) semiflexible DNA Abschnitte erzeugt. Durch die Wahl des Synthesewegs konnte sowohl die Länge der semiflexiblen Einheit als auch die Länge und die Sequenz des sticky-ends variiert werden. Anhand dieser Modellverbindungen wurde dann das Hybridisierungsverhalten in Abhängigkeit der Linker- und Segmentlängen untersucht.

An interferon regulatory factor element controls the major immune modulator of murine cytomegalovirus

Immune modulation by herpesviruses, such as cytomegalovirus, is critical for the establishment of acute and persistent infection confronting a vigorous antiviral immune response of the host. Therefore, the action of immune-modulatory proteins has long been the subject of research, with the final goal to identify new strategies for antiviral therapy.rnIn the case of murine cytomegalovirus (mCMV), the viral m152 protein has been identified to play a major role in targeting components of both the innate and the adaptive immune system in terms of infected host-cell recognition in the effector phase of the antiviral immune response. On the one hand, it inhibits cell surface expression of RAE-1 and thereby prevents ligation of the activating natural killer (NK)-cell receptor NKG2D. On the other hand, it decreases cell surface expression of peptide-loaded MHC class I molecules thereby preventing antigen presentation to CD8 T cells. Ultimately, the outcome of CMV infection is determined by the interplay between viral and cellular factors.rnIn this context, the work presented here has revealed a novel and intriguing connection between viral m152 and cellular interferon (IFN), a key cytokine of the immune system: rnthe m152 promoter region contains an interferon regulatory factor element (IRFE) perfectly matching the consensus sequence of cellular IRFEs.rnThe biological relevance of this regulatory element was first suggested by sequence comparisons revealing its evolutionary conservation among various established laboratory strains of mCMV and more recent low-passage wild-derived virus isolates. Moreover, search of the mCMV genome revealed only three IRFE sites in the complete sequence. Importantly, the functionality of the IRFE in the m152 promoter was confirmed with the use of a mutant virus, representing a functional deletion of the IRFE, and its corresponding revertant virus. In particular, m152 gene expression was found to be inhibited in an IRFE-dependent manner in infected cells. Essentially, this inhibition proved to have a severe impact on the immune-modulatory function of m152, first demonstrated by a restored direct antigen presentation on infected cells for CD8 T-cell activation. Even more importantly, this effect of IRFE-mediated IFN signaling was validated in vivo by showing that the protective antiviral capacity of adoptively-transferred, antigen-specific CD8 T cells is also significantly restored by the IRFE-dependent inhibition of m152. Somewhat curious and surprising, the decrease in m152 protein simultaneously prevented an enhanced activation of NK cells in acute-infected mice, apparently independent of the RAE-1/NKG2D ligand/receptor interaction but rather due to reduced ‘missing-self’ recognition.rnTaken together, this work presents a so far unknown mechanism of IFN signaling to control mCMV immune modulation in acute infection.rnrn

The specificity of the secondary DNA binding site of RecA protein defines its role in DNA strand exchange.

The RecA protein-single-stranded DNA (ssDNA) filament can bind a second DNA molecule. Binding of ssDNA to this secondary site shows specificity, in that polypyrimidinic DNA binds to the RecA protein-ssDNA filament with higher affinity than polypurinic sequences. The affinity of ssDNA, which is identical in sequence to that bound in the primary site, is not always greater than that of nonhomologous DNA. Moreover, this specificity of DNA binding does not depend on the sequence of the DNA bound to the RecA protein primary site. We conclude that the specificity reflects an intrinsic property of the secondary site of RecA protein rather than an interaction between DNa molecules within nucleoprotein filament--i.e., self-recognition. The secondary DNA binding site displays a higher affinity for ssDNA than for double-stranded DNA, and the binding of ssDNA to the secondary site strongly inhibits DNA strand exchange. We suggest that the secondary binding site has a dual role in DNA strand exchange. During the homology search, it binds double-stranded DNA weakly; upon finding local homology, this site binds, with higher affinity, the ssDNA strand that is displaced during DNA strand exchange. These characteristics facilitate homologous pairing, promote stabilization of the newly formed heteroduplex DNA, and contribute to the directionality of DNA strand exchange.

Gestational drive and the green-bearded placenta.

A "green beard" refers to a gene, or group of genes, that is able to recognize itself in other individuals and direct benefits to these individuals. Green-beard effects have been dismissed as implausible by authors who have implicitly assumed sophisticated mechanisms of perception and complex behavioral responses. However, many simple mechanisms for genes to "recognize" themselves exist at the maternal-fetal interface of viviparous organisms. Homophilic cell adhesion molecules, for example, are able to interact with copies of themselves on other cells. Thus, the necessary components of a green-beard effect -- feature, recognition, and response -- can be different aspects of the phenotype of a single gene. Other green-beard effects could involve coalitions of genes at closely linked loci. In fact, any form of epistasis between a locus expressed in a mother and a closely linked locus expressed in the fetus has the property of "self-recognition." Green-beard effects have many formal similarities to systems of meiotic drive and, like them, can be a source of intragenomic conflict.

T cells signaled by NF-kappa B- dendritic cells are sensitized not anergic to subsequent activation

Paradoxically, while peripheral self-tolerance exists for constitutively presented somatic self Ag, self-peptide recognized in the context of MHC class II has been shown to sensitize T cells for subsequent activation. We have shown that MHC class II(+)CD86(+)CD40(-) DC, which can be generated from bone marrow in the presence of an NF-kappaB inhibitor, and which constitutively populate peripheral tissues and lymphoid organs in naive animals, can induce Ag-specific tolerance. In this study, we show that CD40(-) human monocyte-derived dendritic cells (DC), generated in the presence of an NF-kappaB inhibitor, signal phosphorylation of TCRzeta, but little proliferation or IFN-gamma in vitro. Proliferation is arrested in the G(1)/G(0) phase of the cell cycle. Surprisingly, responding T cells are neither anergic nor regulatory, but are sensitized for subsequent IFN-gamma production. The data indicate that signaling through NF-kappaB determines the capacity of DC to stimulate T cell proliferation. Functionally, NF-kappaB(-)CD40(-)class II+ DC may either tolerize or sensitize T cells. Thus, while CD40(-) DC appear to prime or prepare T cells, the data imply that signals derived from other cells drive the generation either of Ag-specific regulatory or effector cells in vivo.

Generic Technique to Generate Large Branched DNA Complexes

The inherent self-recognition properties of DNA have led to its use as a scaffold for various nanotechnology self-assembly applications, with macromolecular complexes, metallic and semiconducting nanoparticles, proteins, inter alia, being assembled onto a designed DNA scaffold. Such structures may typically comprise a number of DNA molecules organized into macromolecules. Many studies have used synthetic methods to produce the constituent DNA molecules, but this typically constrains the molecules to be no longer than around 100 base pairs (30 nm). However, applications that require larger self-assembling DNA complexes, several tens of nanometers or more, need to be generated by other techniques. Here, we present a generic technique to generate large linear, branched, and/or circular DNA macromolecular complexes. The effectiveness of this technique is demonstrated here by the use of Lambda Bacteriophage DNA as a template to generate single- and double-branched DNA structures approximately 120 nm in size.

Analysis of Heterokaryon Formation in Fusarium oxysporum f. sp. cubense.

Fusarium oxysporum forma specialis cubense is a soilborne phytopathogen that infects banana. The true evolutionary identity of this so called species, Fusarium oxysporum, is still unknown. Many techniques have been applied in order to gain insight for the observed genetic diversity of this species. The current classification system is based on vegetative compatibility groups (VCG's). Vegetative compatibility is a self non-self recognition system in which only those belonging to a VCG can form stable heterokaryons, cells containing two distinct nuclei. Heterokaryons in turn, are formed from hypha! anastomosis, the fusion of two hyphae. Furthermore, subsequent to heterokaryon formation potential mechanisms exist which may generate genetic variability. One is through viral transfer upon hyphal anastomosis. The other mechanism is a form of mitotic recombination referred to as the parasexual cycle. Very little research has been performed to directly obser.ve the cellular events; hypha! anastomosis, heterokaryon formation, and the parasexual cycle in Fusarium oxysporum f. sp. cubense. The purpose of this research was to design and use methods which would allow for the detection of hypha! anastomosis and heterokaryon formation, as well as any characteristics surrounding this event, within and between VCG's in Foe. First, some general growth properties were recorded: the number of nuclei per hypha, the size ofthe hyphal tip cell, the size of the cell adjacent to the hypha! tip (pre-tip) cell, and the number of cells to the first branch point. Second, four methods were designed in order to assay hyphal anastomosis and heterokaryon formation: 1) pairings on membrane: phase or brightfield microscopy, 2) pairings on membrane: fluorescence microscopy, 3) spore crosses: fluorescence microscopy, and 4) double picks in fractionated MMA. All of these methods were promtsmg.

Two “pillars” of cold-water coral reefs along Atlantic European margins: Prevalent association of Madrepora oculata with Lophelia pertusa, from reef to colony scale

The scleractinian coral Lophelia pertusa has been the focus of deep-sea research since the recognition of the vast extent of coral reefs in North Atlantic waters two decades ago, long after their existence was mentioned by fishermen. These reefs where shown to provide habitat, concentrate biomass and act as feeding or nursery grounds for many species, including those targeted by commercial fisheries. Thus, the attention given to this cold-water coral (CWC) species from researchers and the wider public has increased. Consequently, new research programs triggered research to determine the full extent of the corals geographic distribution and ecological dynamics of “Lophelia reefs”. The present study is based on a systematic standardised sampling design to analyse the distribution and coverage of CWC reefs along European margins from the Bay of Biscay to Iceland. Based on Remotely Operated Vehicle (ROV) image analysis, we report an almost systematic occurrence of Madrepora oculata in association with L. pertusa with similar abundances of both species within explored reefs, despite a tendency of increased abundance of L. pertusa compared to M. oculata toward higher latitudes. This systematic association occasionally reached the colony scale, with “twin” colonies of both species often observed growing next to each other when isolated structures were occurring off-reefs. Finally, several “false chimaera” were observed within reefs, confirming that colonial structures can be “coral bushes” formed by an accumulation of multiple colonies even at the inter-specific scale, with no need for self-recognition mechanisms. Thus, we underline the importance of the hitherto underexplored M. oculata in the Eastern Atlantic, re-establishing a more balanced view that both species and their yet unknown interactions are required to better elucidate the ecology, dynamics and fate of European CWC reefs in a changing environment.