Interface cena e tecnologia : composições cênicas mediadas

Based on the presupposition that the arts in the West always counted on resources, supports, and devices pertaining to its time context, an reflection is intended regarding the scenic compositions mediated by digital technologies do. Such technologies are inserted in the daily routine, also composing artistic experiments, thus playing a dialogical role with the art/technology intersection. Therefore, the proposal is to investigate what relationships are established in the contemporary theatrical scene from the contagion by digital technologies, aiming at establishing this parallel through a dialogue with the authors discussing the subject, and also based on the group practices having technological resources as a determinant factor in their plays. Furthermore, a reflection should be made on the scene that incorporates or is carried out in intermediatic events, analyzing how digital technologies (re)configure compositional processes of the plays by GAG Phila7, in the city of São Paulo/SP. For such, the dissertation is organized in three sections comprising four moments, to wit: brief overview of the field, contextualization, poetic analysis and synthesis. Qualitative methods are used as the methodological proposal: semi-structure interview, note and document taking (program, website, playing book, disclosure material for advertising text, photographs, and videos). Within the universe of qualitative research, it works with the epistemological perspective of the Gadamer philosophical hermeneutics. The possibilities allowed by the double virtual (Internet/web) generated a type of theater with another material basis and new forms of organization and structure, being possible to perceive that such technological advances and the arts are mutually contaminated, generating a dislocation in the logics of theatrical composition, movement beginning with the artistic vanguards, gradually intensified, thus offering new possibilities of constructions and hybridization of the of the most different possible types. Experiment ―Profanações_superfície de eventos de construção coletiva‖, idealized by Phila7 is inserted in this perspective. Object of the discussion of such research, the experiment works with possible poetics arising from the intersection with the digital technologies, aiming at identifying and problematizing the challenges from the technological evolution and expansion in a scenic context

Interface cena e tecnologia : composições cênicas mediadas

Based on the presupposition that the arts in the West always counted on resources, supports, and devices pertaining to its time context, an reflection is intended regarding the scenic compositions mediated by digital technologies do. Such technologies are inserted in the daily routine, also composing artistic experiments, thus playing a dialogical role with the art/technology intersection. Therefore, the proposal is to investigate what relationships are established in the contemporary theatrical scene from the contagion by digital technologies, aiming at establishing this parallel through a dialogue with the authors discussing the subject, and also based on the group practices having technological resources as a determinant factor in their plays. Furthermore, a reflection should be made on the scene that incorporates or is carried out in intermediatic events, analyzing how digital technologies (re)configure compositional processes of the plays by GAG Phila7, in the city of São Paulo/SP. For such, the dissertation is organized in three sections comprising four moments, to wit: brief overview of the field, contextualization, poetic analysis and synthesis. Qualitative methods are used as the methodological proposal: semi-structure interview, note and document taking (program, website, playing book, disclosure material for advertising text, photographs, and videos). Within the universe of qualitative research, it works with the epistemological perspective of the Gadamer philosophical hermeneutics. The possibilities allowed by the double virtual (Internet/web) generated a type of theater with another material basis and new forms of organization and structure, being possible to perceive that such technological advances and the arts are mutually contaminated, generating a dislocation in the logics of theatrical composition, movement beginning with the artistic vanguards, gradually intensified, thus offering new possibilities of constructions and hybridization of the of the most different possible types. Experiment ―Profanações_superfície de eventos de construção coletiva‖, idealized by Phila7 is inserted in this perspective. Object of the discussion of such research, the experiment works with possible poetics arising from the intersection with the digital technologies, aiming at identifying and problematizing the challenges from the technological evolution and expansion in a scenic context

Interface cena e tecnologia : composições cênicas mediadas

Based on the presupposition that the arts in the West always counted on resources, supports, and devices pertaining to its time context, an reflection is intended regarding the scenic compositions mediated by digital technologies do. Such technologies are inserted in the daily routine, also composing artistic experiments, thus playing a dialogical role with the art/technology intersection. Therefore, the proposal is to investigate what relationships are established in the contemporary theatrical scene from the contagion by digital technologies, aiming at establishing this parallel through a dialogue with the authors discussing the subject, and also based on the group practices having technological resources as a determinant factor in their plays. Furthermore, a reflection should be made on the scene that incorporates or is carried out in intermediatic events, analyzing how digital technologies (re)configure compositional processes of the plays by GAG Phila7, in the city of São Paulo/SP. For such, the dissertation is organized in three sections comprising four moments, to wit: brief overview of the field, contextualization, poetic analysis and synthesis. Qualitative methods are used as the methodological proposal: semi-structure interview, note and document taking (program, website, playing book, disclosure material for advertising text, photographs, and videos). Within the universe of qualitative research, it works with the epistemological perspective of the Gadamer philosophical hermeneutics. The possibilities allowed by the double virtual (Internet/web) generated a type of theater with another material basis and new forms of organization and structure, being possible to perceive that such technological advances and the arts are mutually contaminated, generating a dislocation in the logics of theatrical composition, movement beginning with the artistic vanguards, gradually intensified, thus offering new possibilities of constructions and hybridization of the of the most different possible types. Experiment ―Profanações_superfície de eventos de construção coletiva‖, idealized by Phila7 is inserted in this perspective. Object of the discussion of such research, the experiment works with possible poetics arising from the intersection with the digital technologies, aiming at identifying and problematizing the challenges from the technological evolution and expansion in a scenic context

Abrogation of contaminating RNA activity in HIV-1 Gag VLPs

Background: HIV-1 Gag virus like particles (VLPs) used as candidate vaccines are regarded as inert particles as they contain no replicative nucleic acid, although they do encapsidate cellular RNAs. During HIV-1 Gag VLP production in baculovirus-based expression systems, VLPs incorporate the baculovirus Gp64 envelope glycoprotein, which facilitates their entry into mammalian cells. This suggests that HIV-1 Gag VLPs produced using this system facilitate uptake and subsequent expression of encapsidated RNA in mammalian cells - an unfavourable characteristic for a vaccine. Methods. HIV-1 Gag VLPs encapsidating reporter chloramphenicol acetyl transferase (CAT) RNA, were made in insect cells using the baculovirus expression system. The presence of Gp64 on the VLPs was verified by western blotting and RT-PCR used to detect and quantitate encapsidated CAT RNA. VLP samples were heated to inactivate CAT RNA. Unheated and heated VLPs incubated with selected mammalian cell lines and cell lysates tested for the presence of CAT protein by ELISA. Mice were inoculated with heated and unheated VLPs using a DNA prime VLP boost regimen. Results: HIV-1 Gag VLPs produced had significantly high levels of Gp64 (∼1650 Gp64 molecules/VLP) on their surfaces. The amount of encapsidated CAT RNA/g Gag VLPs ranged between 0.1 to 7 ng. CAT protein was detected in 3 of the 4 mammalian cell lines incubated with VLPs. Incubation with heated VLPs resulted in BHK-21 and HeLa cell lysates showing reduced CAT protein levels compared with unheated VLPs and HEK-293 cells. Mice inoculated with a DNA prime VLP boost regimen developed Gag CD8 and CD4 T cell responses to GagCAT VLPs which also boosted a primary DNA response. Heating VLPs did not abrogate these immune responses but enhanced the Gag CD4 T cell responses by two-fold. Conclusions: Baculovirus-produced HIV-1 Gag VLPs encapsidating CAT RNA were taken up by selected mammalian cell lines. The presence of CAT protein indicates that encapsidated RNA was expressed in the mammalian cells. Heat-treatment of the VLPs altered the ability of protein to be expressed in some cell lines tested but did not affect the ability of the VLPs to stimulate an immune response when inoculated into mice. © 2011 Valley-Omar et al; licensee BioMed Central Ltd.

Human immunodeficiency virus type 1 subtype C Gag virus-like particle boost substantially improves the immune response to a subtype C gag DNA vaccine in mice

Human immunodeficiency virus type 1 (HIV-1) subtype C is the predominant HIV in southern Africa, and is the target of a number of recent vaccine candidates. It has been proposed that a heterologous prime/boost vaccination strategy may result in stronger, broader and more prolonged immune responses. Since HIV-1 Gag Pr55 polyprotein can assemble into virus-like particles (VLPs) which have been shown to induce a strong cellular immune response in animals, we showed that a typical southern African subtype C Pr55 protein expressed in insect cells via recombinant baculovirus could form VLPs. We then used the baculovirus-produced VLPs as a boost to a subtype C HIV-1 gag DNA prime vaccination in mice. This study shows that a low dose of HIV-1 subtype C Gag VLPs can significantly boost the immune response to a single subtype C gag DNA inoculation in mice. These results suggest a possible vaccination regimen for humans. © 2004 SGM.

Chimaeric HIV-1 subtype C Gag molecules with large in-frame C-terminal polypeptide fusions form virus-like particles

HIV-1 Pr55 Gag virus-like particles (VLPs) are strong immunogens with potential as candidate HIV vaccines. VLP immunogenicity can be broadened by making chimaeric Gag molecules: however, VLPs incorporating polypeptides longer than 200 aa fused in frame with Gag have not yet been reported. We constructed a range of gag-derived genes encoding in-frame C-terminal fusions of myristoylation-competent native Pr55Gag and p6-truncated Gag (Pr50Gag) to test the effects of polypeptide length and sequence on VLP formation and morphology, in an insect cell expression system. Fused sequences included a modified reverse transcriptase-Tat-Nef fusion polypeptide (RTTN, 778 aa), and truncated versions of RTTN ranging from 113 aa to 450 aa. Baculovirus-expressed chimaeric proteins were examined by western blot and electron microscopy. All chimaeras formed VLPs which could be purified by sucrose gradient centrifugation. VLP diameter increased with protein MW, from ∼100 nm for Pr55Gag to ∼250 nm for GagRTTN. The presence or absence of the Gag p6 region did not obviously affect VLP formation or appearance. GagRT chimaeric particles were successfully used in mice to boost T-cell responses to Gag and RT that were elicited by a DNA vaccine encoding a GagRTTN polypeptide, indicating the potential of such chimaeras to be used as candidate HIV vaccines. © 2008 Elsevier B.V. All rights reserved.

A prime-boost immunisation regimen using recombinant BCG and Pr55 gag virus-like particle vaccines based on HIV type 1 subtype C successfully elicits Gag-specific responses in baboons

Mycobacterium bovis BCG is considered an attractive live bacterial vaccine vector. In this study, we investigated the immune response of baboons to a primary vaccination with recombinant BCG (rBCG) constructs expressing the gag gene from a South African HIV-1 subtype C isolate, and a boost with HIV-1 subtype C Pr55 gag virus-like particles (Gag VLPs). Using an interferon enzyme-linked immunospot assay, we show that although these rBCG induced only a weak or an undetectable HIV-1 Gag-specific response on their own, they efficiently primed for a Gag VLP boost, which strengthened and broadened the immune responses. These responses were predominantly CD8+ T cell-mediated and recognised similar epitopes as those targeted by humans with early HIV-1 subtype C infection. In addition, a Gag-specific humoral response was elicited. These data support the development of HIV-1 vaccines based on rBCG and Pr55 gag VLPs. © 2009 Elsevier Ltd. All rights reserved.

Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production

Background Insect baculovirus-produced Human immunodeficiency virus type 1 (HIV-1) Gag virus-like-particles (VLPs) stimulate good humoral and cell-mediated immune responses in animals and are thought to be suitable as a vaccine candidate. Drawbacks to this production system include contamination of VLP preparations with baculovirus and the necessity for routine maintenance of infectious baculovirus stock. We used piggyBac transposition as a novel method to create transgenic insect cell lines for continuous VLP production as an alternative to the baculovirus system. Results Transgenic cell lines maintained stable gag transgene integration and expression up to 100 cell passages, and although the level of VLPs produced was low compared to baculovirus-produced VLPs, they appeared similar in size and morphology to baculovirus-expressed VLPs. In a murine immunogenicity study, whereas baculovirus-produced VLPs elicited good CD4 immune responses in mice when used to boost a prime with a DNA vaccine, no boost response was elicited by transgenically produced VLPs. Conclusion Transgenic insect cells are stable and can produce HIV Pr55 Gag VLPs for over 100 passages: this novel result may simplify strategies aimed at making protein subunit vaccines for HIV. Immunogenicity of the Gag VLPs in mice was less than that of baculovirus-produced VLPs, which may be due to lack of baculovirus glycoprotein incorporation in the transgenic cell VLPs. Improved yield and immunogenicity of transgenic cell-produced VLPs may be achieved with the addition of further genetic elements into the piggyBac integron.

Conserved and variable features of gag structure and arrangement in immature retrovirus particles

The assembly of retroviruses is driven by oligomerization of the Gag polyprotein. We have used cryo-electron tomography together with subtomogram averaging to describe the three-dimensional structure of in vitro-assembled Gag particles from human immunodeficiency virus, Mason-Pfizer monkey virus, and Rous sarcoma virus. These represent three different retroviral genera: the lentiviruses, betaretroviruses and alpharetroviruses. Comparison of the three structures reveals the features of the supramolecular organization of Gag that are conserved between genera and therefore reflect general principles of Gag-Gag interactions and the features that are specific to certain genera. All three Gag proteins assemble to form approximately spherical hexameric lattices with irregular defects. In all three genera, the N-terminal domain of CA is arranged in hexameric rings around large holes. Where the rings meet, 2-fold densities, assigned to the C-terminal domain of CA, extend between adjacent rings, and link together at the 6-fold symmetry axis with a density, which extends toward the center of the particle into the nucleic acid layer. Although this general arrangement is conserved, differences can be seen throughout the CA and spacer peptide regions. These differences can be related to sequence differences among the genera. We conclude that the arrangement of the structural domains of CA is well conserved across genera, whereas the relationship between CA, the spacer peptide region, and the nucleic acid is more specific to each genus.