934 resultados para Particularly and concrete administrative act
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La imagen fotográfica es un bloque espacio tiempo congelado, un fragmento referido al antes y el después de algo. Al contemplar una fotografía de un interior doméstico, descubrimos un entretejido sutil entre el habitante y su hábitat. Somos capaces de recaer en más detalles de los que el ojo humano puede apreciar en su visión cotidiana, siempre ligada al devenir espacio temporal. El acto de fotografiar el hogar, de congelar unidades habitadas infinitesimales, se revela como una manifestación radical del modo unipersonal de habitar de cada fotógrafo, profesional o aficionado, y por extensión, dado que hoy todos somos fotógrafos; de cada habitante. Por un lado, la fotografía se piensa aquí como herramienta, capaz de desvelar, de poner en el mundo, los elementos, percepciones y acontecimientos, que subyacen imbricados en la construcción del hogar. Por otro, la imagen se entiende como medio de expresión y de comunicación, como el lenguaje universal de nuestro tiempo, por todos conocido y utilizado. En este momento de interconexión máxima, entre redes, datos y capas de cognición, de velocidad y aceleración, esta tesis doctoral se plantea como una vuelta a la reflexión, a la contemplación del objeto imagen, desde la certeza de que para que ésta hable hay que darle tiempo. Así, la investigación hay que entenderla desde una base ontológica y fenomenológica; desde la experiencia del ser que habita un entorno concreto y determinado. Se enmarca en el actual entorno socio cultural de occidente, se busca desvelar el significado y modo de habitar del habitante común, poniendo de manifiesto aquello que acontece para que una casa cualquiera, de un habitante cualquiera, devenga hogar. Los primeros indicios sobre el tema surgirán del análisis y la reinterpretación hermenéutica de un atlas de imágenes del habitar: cuerpo de imágenes reunido a partir de series fotográficas de hogares, de habitantes anónimos, puestos a luz por la mirada de un grupo de artistas. Posteriormente, ponemos a prueba el conocimiento adquirido en el análisis anterior, mientras que expandimos la investigación hacia el sentir del habitante común, mediante la realización de tres experimentos participativos, o estudios de campo cualitativos. Los resultados, de ambos grupos de casos de estudio, se compilan, organizan y estructuran en una taxonomía del habitar. Esta taxonomía está compuesta por cuarenta y siete parámetros, que explicitan la naturaleza compleja del hogar del siglo XXI. Este hogar es entendido como un constructo personal de cada habitante, un proceso que acontece en el tiempo y en el espacio, y que entraña la propia construcción del habitante. Esta extensa taxonomía se organiza según tres ámbitos del ser humano, en el primero se expresan los factores relacionados con el modo de "estar físicamente" en el hogar, incluyendo: al propio habitante, la casa como espacio arquitectónico y como materialidad: objetos, muebles, iconos y símbolos que pueblan el hogar. En segundo lugar, se manifiestan los parámetros relacionados con el modo de “percibir”: por un lado, aquello que se deriva de lo que se ve, y por otro, lo que se deriva de aquello que no se ve, pero se siente. En tercer lugar, se explicitan los factores relativos al habitante que "crea/juega" su hogar, quién por un lado, es en el mundo actuando, y que por otro, siente el mundo construyéndolo mediante una serie de relaciones que establece con él. Finalmente, la investigación intenta revelar las sinergias, conexiones y relaciones, entre todos estos elementos extraídos del sentir del habitante común, y que fueron inducidos mediante el análisis y reinterpretación de los casos de estudio, poniendo de manifiesto un orden de cosas en el habitar occidental contemporáneo. ABSTRACT The photographic image is a frozen space time block, a fragment referred to a something before and after. When we stare at the photography of domestic interiors we discover a subtle interweaving between the inhabitant and her habitat. We are able to acknowledge infinite more details than the human eye, in its continuous quotidian vision always linked to the space time progression, appreciates. The act of photographing the home, of freezing infinitesimal inhabited units, reveals as a radical statement of the concept of inhabiting for each photographer, professional or amateur, and by extension, as we today all are photographers, for each inhabitant. On the one hand, photography is here conceived as a tool that is capable of revealing, "of placing in the world" the elements, perceptions and happenings that underlie imbricated in the construction of a home. On the other hand, image is thought as an expression and communication media, as the universal language of our time (as far as it is known and used by all of us). In this precise moment of maximum interconnection between networks, data and cognitive layers; of speed and acceleration, this PhD Dissertation is conceived as a return to reflection; to the contemplation of object image, from the certainty of its need of time for talking. Therefore, this research from an ontological and phenomenological base; from the experience of the self who inhabits a determined and concrete environment, that of the western countries at the present, pursues to unveil the meaning and way of inhabiting of a common dweller and manifest what conforms the transformation of any house, of any inhabitant into a home. The first clues will arise from the analysis and hermeneutical reinterpretation of the Atlas of inhabiting; an assembled body of images of anonymous inhabitants houses, brought into life through a group of artist´s glance. Afterwards, we will test the analysis´s acquired knowledge, while extending the investigation to the feel of the common inhabitant (and no longer the expert^ artist) through the execution of three participative experiments conceived as qualitative field works. The results of both case study groups, will be compiled, organized and structured in a taxonomy of the inhabiting. This taxonomy is composed by forty seven parameters that explicitly state the complex nature of the XXI century home, regarded as a personal construct of every single inhabitant, as a process that happens through time and space and that entails the construction of the inhabitant. This wide taxonomy is organized regarding three spheres of the human being, In first place, those elements related to the way of “physically being” at home are expressed, including: the inhabitant its self, the house as architectural space and as materiality: objects, furniture, icons and symbols that fill the home. In second place, parameters related to the way of “perceiving“ are manifested; on the one hand, those that derive from what we see; on the other hand, those that derive from what we do not see, but feel. In third place, those factors deriving from the inhabitant as a home "creator/player" who is acting in the world and feeling the world while constructing it through a myriad of relationships he establishes with it. Finally, the investigation tries to reveal the synergies, connections and relations between all these elements extracted from the feelings of the common inhabitant, induced through the analysis and reinterpretation of the case studies, and therefore exposing a state of things belonging to western world at present.
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A cultura de uma organização é importante para que seus colaboradores possuam mesmos objetivos e valores. Porém, em busca de manter uma estratégia competitiva e agir de forma responsável na comunidade em que se encontra, a empresa precisa inovar e adaptar-se. A gestão da diversidade apresenta-se como uma válida forma de enfrentar estes novos desafios e exigências. Contudo, há muitos obstáculos para que uma gestão da diversidade seja bem-sucedida. Este estudo propõe-se em entender como a diversidade pode afetar a cultura de uma organização. Além disso, como a cultura pode afetar a estratégia de gestão da diversidade. Foi utilizado para este objetivo um estudo de caso em profundidade com seis entrevistas. As entrevistas foram apoiadas em um roteiro semi estruturado. A análise dos dados obtidos foi feita pela análise de conteúdo. De acordo com a pesquisa realizada, sugere que a cultura organizacional e a gestão da diversidade estão diretamente conectadas e que podem influenciar positivamente uma a outra, porém precisam manter um equilíbrio em suas ações para que não causem prejuízos à organização.
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A universal base that is capable of substituting for any of the four natural bases in DNA would be of great utility in both mutagenesis and recombinant DNA experiments. This paper describes the properties of oligonucleotides incorporating two degenerate bases, the pyrimidine base 6H,8H-3,4-dihydropyrimido[4,5-c][1,2]oxazin-7-one and the purine base N6-methoxy-2,6-diaminopurine, designated P and K, respectively. An equimolar mixture of the analogues P and K (called M) acts, in primers, as a universal base. The thermal stability of oligonucleotide duplexes were only slightly reduced when natural bases were replaced by P or K. Templates containing the modified bases were copied by Taq polymerase; P behaved as thymine in 60% of copying events and as cytosine in 40%, whereas K behaved as if it were guanine (13%) or adenine (87%). The dUTPase gene of Caenorhabditis elegans, which we have found to contain three nonidentical homologous repeats, was used as a model system to test the use of these bases in primers for DNA synthesis. A pair of oligodeoxyribonucleotides, each 20 residues long and containing an equimolar mixture of P and K at six positions, primed with high specificity both T7 DNA polymerase in sequencing reactions and Taq polymerase in PCRs; no nonspecific amplification was obtained on genomic DNA of C. elegans. Use of P and K can significantly reduce the complexity of degenerate oligonucleotide mixtures, and when used together, P and K can act as a universal base.
Masking and unmasking of the sialic acid-binding lectin activity of CD22 (Siglec-2) on B lymphocytes
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CD22 is a B cell-restricted glycoprotein involved in signal transduction and modulation of cellular activation. It is also an I-type lectin (now designated Siglec-2), whose extracellular domain can specifically recognize α2–6-linked sialic acid (Sia) residues. This activity is postulated to mediate intercellular adhesion and/or to act as a coreceptor in antigen-induced B cell activation. However, studies with recombinant CD22 indicate that the lectin function can be inactivated by expression of α2–6-linked Sia residues on the same cell surface. To explore whether this masking phenomenon affects native CD22 on B cells, we first developed a probe to detect the lectin activity of recombinant CD22 expressed on Chinese hamster ovary cells (which have no endogenous α2–6-linked Sia residues). This probe is inactive against CD22-positive B lymphoma cells and Epstein–Barr virus-transformed lymphoblasts which express high levels of α2–6-linked Sia residues. Enzymatic desialylation unmasks the CD22 lectin activity, indicating that endogenous Sia residues block the CD22 lectin-binding site. Truncation of the side chains of cell surface Sia residues by mild periodate oxidation (known to abrogate Sia recognition by CD22) also had this unmasking effect, indicating that the effects of desialylation are not due to a loss of negative charge. Normal resting B cells from human peripheral blood gave similar findings. However, the lectin is partially unmasked during in vitro activation of these cells. Thus, the lectin activity of CD22 is restricted by endogenous sialylation in resting B cells and may be transiently unmasked during in vivo activation, perhaps to modulate intercellular or intracellular interactions at this critical stage in the humoral response.
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We thank Karim Gharbi and Urmi Trivedi for their assistance with RNA sequencing, carried out in the GenePool genomics facility (University of Edinburgh). We also thank Susan Fairley and Eduardo De Paiva Alves (Centre for Genome Enabled Biology and Medicine, University of Aberdeen) for help with the initial bioinformatics analysis. We thank Aaron Mitchell for kindly providing the ALS3 mutant, Julian Naglik for the gift of TR146 cells, and Jon Richardson for technical assistance. We thank the Genomics and Bioinformatics core of the Faculty of Health Sciences for Next Generation Sequencing and Bioinformatics support, the Information and Communication Technology Office at the University of Macau for providing access to a High Performance Computer and Jacky Chan and William Pang for their expert support on the High Performance Computer. Finally, we thank Amanda Veri for generating CaLC2928. M.D.L. is supported by a Sir Henry Wellcome Postdoctoral Fellowship (Wellcome Trust 096072), R.A.F. by a Wellcome Trust-Massachusetts Institute of Technology (MIT) Postdoctoral Fellowship, L.E.C. by a Canada Research Chair in Microbial Genomics and Infectious Disease and by Canadian Institutes of Health Research Grants MOP-119520 and MOP-86452, A.J. P.B. was supported by the UK Biotechnology and Biological Sciences Research Council (BB/F00513X/1) and by the European Research Council (ERC-2009-AdG-249793-STRIFE), KHW is supported by the Science and Technology Development Fund of Macau S.A.R (FDCT) (085/2014/A2) and the Research and Development Administrative Office of the University of Macau (SRG2014-00003-FHS) and R.T.W. by the Burroughs Wellcome fund and NIH R15AO094406. Data availability RNA-sequencing data sets are available at ArrayExpress (www.ebi.ac.uk) under accession code E-MTAB-4075. ChIP-seq data sets are available at the NCBI SRA database (http://www.ncbi.nlm.nih.gov) under accession code SRP071687. The authors declare that all other data supporting the findings of this study are available within the article and its supplementary information files, or from the corresponding author upon request.
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Assembly and mutual proximities of α, β, and γc subunits of the interleukin 2 receptors (IL-2R) in plasma membranes of Kit 225 K6 T lymphoma cells were investigated by fluorescence resonance energy transfer (FRET) using fluorescein isothiocyanate- and Cy3-conjugated monoclonal antibodies (mAbs) that were directed against the IL-2Rα, IL-2Rβ, and γc subunits of IL-2R. The cell-surface distribution of subunits was analyzed at the nanometer scale (2–10 nm) by FRET on a cell-by-cell basis. The cells were probed in resting phase and after coculture with saturating concentrations of IL-2, IL-7, and IL-15. FRET data from donor- and acceptor-labeled IL-2Rβ-α, γ-α, and γ-β pairs demonstrated close proximity of all subunits to each other in the plasma membrane of resting T cells. These mutual proximities do not appear to represent mAb-induced microaggregation, because FRET measurements with Fab fragments of the mAbs gave similar results. The relative proximities were meaningfully modulated by binding of IL-2, IL-7, and IL-15. Based on FRET analysis the topology of the three subunits at the surface of resting cells can be best described by a “triangular model” in the absence of added interleukins. IL-2 strengthens the bridges between the subunits, making the triangle more compact. IL-7 and IL-15 act in the opposite direction by opening the triangle possibly because they associate their private specific α receptors with the β and/or γc subunits of the IL-2R complex. These data suggest that IL-2R subunits are already colocalized in resting T cells and do not require cytokine-induced redistribution. This colocalization is significantly modulated by binding of relevant interleukins in a cytokine-specific manner.
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The P1 partition system promotes faithful plasmid segregation during the Escherichia coli cell cycle. This system consists of two proteins, ParA and ParB, that act on a plasmid site called parS. By immunofluorescence microscopy, we observed that ParB localizes to discrete foci that are most often located close to the one-quarter and three-quarters positions of cell length. The visualization of ParB foci depended completely on the presence of parS, although their visualization was independent of the chromosomal context of parS (in P1 or the bacterial chromosome). In integration host factor-defective mutants, in which ParB binding to parS is weakened, only a fraction of the total pool of ParB had converged into foci. Taken together, these results indicate that parS recruits a pool of ParB into foci and that the resulting ParB–parS complexes serve as substrates for the segregation reaction. In the absence of ParA, the position of ParB foci in cells is perturbed, indicating that at least one of the roles of ParA is to direct ParB–parS complexes to the proper one-quarter positions from a cell pole. Finally, inhibition of cell division did not inhibit localization of ParB foci in cells, indicating that the positioning signals in the E. coli host that are needed for P1 partition do not depend on early division events.
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Rho, a member of the Rho small G protein family, regulates the formation of stress fibers and focal adhesions in various types of cultured cells. We investigated here the actions of ROCK and mDia, both of which have been identified to be putative downstream target molecules of Rho, in Madin–Darby canine kidney cells. The dominant active mutant of RhoA induced the formation of parallel stress fibers and focal adhesions, whereas the dominant active mutant of ROCK induced the formation of stellate stress fibers and focal adhesions, and the dominant active mutant of mDia induced the weak formation of parallel stress fibers without affecting the formation of focal adhesions. In the presence of C3 ADP-ribosyltransferase for Rho, the dominant active mutant of ROCK induced the formation of stellate stress fibers and focal adhesions, whereas the dominant active mutant of mDia induced only the diffuse localization of actin filaments. These results indicate that ROCK and mDia show distinct actions in reorganization of the actin cytoskeleton. The dominant negative mutant of either ROCK or mDia inhibited the formation of stress fibers and focal adhesions, indicating that both ROCK and mDia are necessary for the formation of stress fibers and focal adhesions. Moreover, inactivation and reactivation of both ROCK and mDia were necessary for the 12-O-tetradecanoylphorbol-13-acetate–induced disassembly and reassembly, respectively, of stress fibers and focal adhesions. The morphologies of stress fibers and focal adhesions in the cells expressing both the dominant active mutants of ROCK and mDia were not identical to those induced by the dominant active mutant of Rho. These results indicate that at least ROCK and mDia cooperatively act as downstream target molecules of Rho in the Rho-induced reorganization of the actin cytoskeleton.
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With use of the yeast two-hybrid system, the proteins RIP and FADD/MORT1 have been shown to interact with the "death domain" of the Fas receptor. Both of these proteins induce apoptosis in mammalian cells. Using receptor fusion constructs, we provide evidence that the self-association of the death domain of RIP by itself is sufficient to elicit apoptosis. However, both the death domain and the adjacent alpha-helical region of RIP are required for the optimal cell killing induced by the overexpression of this gene. By contrast, FADD's ability to induce cell death does not depend on crosslinking. Furthermore, RIP and FADD appear to activate different apoptotic pathways since RIP is able to induce cell death in a cell line that is resistant to the apoptotic effects of Fas, tumor necrosis factor, and FADD. Consistent with this, a dominant negative mutant of FADD, lacking its N-terminal domain, blocks apoptosis induced by RIP but not by FADD. Since both pathways are blocked by CrmA, the interleukin 1 beta converting enzyme family protease inhibitor, these results suggest that FADD and RIP can act along separable pathways that nonetheless converge on a member of the interleukin 1 beta converting enzyme family of cysteine proteases.
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The MADS domain homeotic proteins APETALA1 (AP1), APETALA3 (AP3), PISTILLATA (PI), and AGAMOUS (AG) act in a combinatorial manner to specify the identity of Arabidopsis floral organs. The molecular basis for this combinatorial mode of action was investigated. Immunoprecipitation experiments indicate that all four proteins are capable of interacting with each other. However, these proteins exhibit "partner-specificity" for the formation of DNA-binding dimers; only AP1 homodimers, AG homodimers, and AP3/PI heterodimers are capable of binding to CArG-box sequences. Both the AP3/PI heterodimer and the AP1 or AG homodimers are formed when the three corresponding proteins are present together. The use of chimeric proteins formed by domain swapping indicates that the L region (which follows the MADS box) constitutes a key molecular determinant for the selective formation of DNA-binding dimers. The implications of these results for the ABC genetic model of flower development are discussed.
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The SSN3 and SSN8 genes of Saccharomyces cerevisiae were identified by mutations that suppress a defect in SNF1, a protein kinase required for release from glucose repression. Mutations in SSN3 and SSN8 also act synergistically with a mutation of the MIG1 repressor protein to relieve glucose repression. We have cloned the SSN3 and SSN8 genes. SSN3 encodes a cyclin-dependent protein kinase (cdk) homolog and is identical to UME5. SSN8 encodes a cyclin homolog 35% identical to human cyclin C. SSN3 and SSN8 fusion proteins interact in the two-hybrid system and coimmunoprecipitate from yeast cell extracts. Using an immune complex assay, we detected protein kinase activity that depends on both SSN3 and SSN8. Thus, the two SSN proteins are likely to function as a cdk-cyclin pair. Genetic analysis indicates that the SSN3-SSN8 complex contributes to transcriptional repression of diversely regulated genes and also affects induction of the GAL1 promoter.
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A estratégia do Tratamento Diretamente Observado de Curta Duração para o controle da tuberculose (DOTS) tem sido usada por vários países do mundo especificamente pelos 22 mais afetados pela doença, que contribuem com cerca de 80% de casos, porém, apesar da estratégia demonstrar a redução de casos e aumentar a adesão dos pacientes ao tratamento, e ter sido implementada em Moçambique desde a década de 1980, a doença continua a ser um problema grave de saúde pública no país. Este estudo tem como objetivo central: analisar a transferência da política do DOTS a partir da visão dos gestores centrais, provinciais e distritais, e profissionais de saúde da província de Nampula. Trata-se de um estudo qualitativo que usa a Análise de Discurso de matriz francesa como seu referencial teórico metodológico que busca a compreensão dos sentidos a partir das condições da produção. Participaram deste estudo 15 profissionais de saúde que ocupavam as posições de gestores, médicos e profissionais de enfermagem e/ou técnicos, e atuavam no Programa nacional de controle de Tuberculose em Moçambique com o mínimo de um ano de experiência, nos meses de maio a agosto de 2014, nos níveis central, provincial (Nampula), e distrital (em oito distritos da província de Nampula). Para a coleta de dados usou-se um roteiro de entrevista semi-estruturado que permitiu explorar os sentidos produzidos. O corpus em análise foi constituído a partir das entrevistas transcritas. Para o auxílio da organização dos dados usou-se o sofware Atlis.ti versão 6. Este estudo teve a aprovação da Comissão Nacional de Bioética e da autorização do Ministro da Saúde de Moçambique. Para a análise dos dados foram identificados quatro blocos discursivos: experiências adotadas na implementação e manutenção do DOTS; estratégias adotadas na implementação e manutenção de DOTS; potencialidades, fragilidades de DOTS no controle da tuberculose e; discursos não buscados pelo roteiro: possíveis soluções. Os sentidos produzidos enfatizam dizeres inscritos em formações discursivas que desconsideram a subjetividade do enfermo; de fragilidades do sistema de saúde, falta de recursos humanos, falta de transporte, baixos salários, insuficiência de laboratórios, distâncias longas entre a moradia do paciente e a unidade sanitária. Ainda os entrevistados entendem como potencialidade a participação dos agentes comunitários e da família no tratamento da doença. Conclui-se que para o controle da tuberculose usando a estratégia DOTS o compromisso do governo deve ser pragmático traduzindo-se em ações concretas como o aumento do financiamento das ações de controle de tuberculose, incluindo as pesquisas, formação dos recursos humanos e, sobretudo, atuar sobre os determinantes socias da saúde
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For over a decade, the U.S. military has been engaged in two distinct, yet equally deadly conflicts: Operation Iraqi Freedom (OIF) and Operation Enduring Freedom (OEF). There are many physical and psychological effects of war necessitating the activation and interventions of a myriad of behavioral health professionals. The purpose of the paper was to understand how and if contemporary military culture may work to support or hinder application of an Acceptance and Commitment Therapy (ACT) approach to issues of psychological health among Soldiers. While the empirical research on efficacy with Soldiers is limited, a review of military culture revealed the promotion of rigid rule following, although effective in combat, influences the emotional control agenda and stigma while in garrison. However, empirical research demonstrating the clinical benefits and flexibility of ACT is rapidly emerging with civilian and Veteran populations. Suggested as a prevention technique utilized early in Soldier's training to increase psychological flexibility, ACT appears to demonstrate much promise in ameliorating the psychological consequences of war.
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Novice therapists training in Acceptance and Commitment Therapy (ACT) may encounter challenges in therapy in which their own personal history functions as a barrier to flexible modes of therapeutic engagement with the therapist. From the ACT perspective, counter-therapeutic interpersonal responses may be examined relative to six behavioral sub-processes. It is suggested that the most vulnerable moments for the therapist will involve those in which certain contextual features of therapy pull historical awareness of a painful personal past into relation with the psychological present. This paper hypothesizes that utilizing approaches based in ACT will assist therapists in overcoming these challenges and will illustrate how to approach case formulation and intervention with therapists in training from a functional contextualistic perspective. To begin, the philosophical and theoretical underpinnings of ACT will be outlined in sufficient depth to intellectually ground the model and its therapeutic project. This conceptual foundation will then be brought to applied focus using hypothetical case material, followed by ACT interventions designed to increase clinical flexibility in the given therapeutic scenario. Future research that systematically examines the effectiveness of such methods among therapists is encouraged.
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Homophobia continues to exist in society. Homonegative attitudes are often implicit and can be acquired without direct training, which makes them particularly resistant to change. Relational Frame Theory (RFT) is a behavior analytic account of learning processes and can explain these processes of indirect learning. RFT also suggests therapeutic processes for dismantling stigma using a therapy model named Acceptance and Commitment Therapy (ACT). This paper reviews previous research on traditional multicultural training, and addresses its shortcomings. Specifically, this paper makes the argument that traditional models encourage experiential avoidance and thus further perpetuate the processes that maintain stigma. While a handful of studies have examined stigma interventions using ACT, no ACT studies have been completed specifically on the stigma towards gay and lesbian individuals. This paper concludes with a research proposal for such a study.