Anti-melanoma antibodies from melanoma patients immunized with genetically modified autologous tumor cells: selection of specific antibodies from single-chain Fv fusion phage libraries.

Fusion phage libraries expressing single-chain Fv antibodies were constructed from the peripheral blood lymphocytes of two melanoma patients who had been immunized with autologous melanoma cells transduced the gamma-interferon gene to enhance immunogenicity, in a trial conducted at another institution. Anti-melanoma antibodies were selected from each library by panning the phage against live cultures of the autologous tumor. After two or three rounds of panning, clones of the phage were tested by ELISA for binding to the autologous tumor cells; > 90% of the clones tested showed a strong ELISA reaction, demonstrating the effectiveness of the panning procedure for selecting antimelanoma antibodies. The panned phage population was extensively absorbed against normal melanocytes to enrich for antibodies that react with melanoma cells but not with melanocytes. The unabsorbed phage were cloned, and the specificities of the expressed antibodies were individually tested by ELISA with a panel of cultured human cells. The first tests were done with normal endothelial and fibroblast cells to identify antibodies that do not react, or react weakly, with two normal cell types, indicating some degree of specificity for melanoma cells. The proportion of phage clones expressing such antibodies was approximately 1%. Those phage were further tested by ELISA with melanocytes, several melanoma lines, and eight other tumor lines, including a glioma line derived from glial cells that share a common lineage with melanocytes. The ELISA tests identified three classes of anti-melanoma antibodies, as follows: (i) a melanoma-specific class that reacts almost exclusively with the melanoma lines; (ii) a tumor-specific class that reacts with melanoma and other tumor lines but does not react with the normal melanocyte, endothelial and fibroblast cells; and (iii) a lineage-specific class that reacts with the melanoma lines, melanocytes, and the glioma line but does not react with the other lines. These are rare classes from the immunized patients' repertoires of anti-melanoma antibodies, most of which are relatively nonspecific anti-self antibodies. The melanoma-specific class was isolated from one patient, and the lineage-specific class was isolated from the other patient, indicating that different patients can have markedly different responses to the same immunization protocol. The procedures described here can be used to screen the antibody repertoire of any person with cancer, providing access to an enormous untapped pool of human monoclonal anti-tumor antibodies with clinical and research potential.

Sentidos de família construídos por profissionais de saúde na estratégia de saúde da família

Na construção de políticas sociais atribuiu-se um papel central à família na proteção social e no cuidado com os seus membros. Na saúde, a família assume essa centralidade na Estratégia de Saúde da Família (ESF), sendo compreendida como objeto da atenção em saúde. Essa centralidade se deu a partir de uma pretensão de mudança no modelo assistencial que visasse a integralidade do cuidado e um olhar para as condições de vida como fundamentais no processo saúde-doença. Tais transformações nos sentidos e práticas de saúde têm sido desafiadoras. A partir da perspectiva Construcionista Social, que orienta esse estudo, compreendemos que os sentidos são construídos nas interações entre as pessoas e que esses configuram práticas sociais. Diante deste quadro de desafios na ESF e a partir da perspectiva adotada, temos como objetivo compreender os sentidos produzidos por profissionais de equipes de saúde sobre famílias em contextos de reuniões de discussão de família/caso na ESF, buscando analisar como esses configuram a produção de práticas de cuidado e também como se dá a dinâmica de construção desses sentidos a partir das negociações entre os participantes. A constituição do corpus foi feita a partir da observação e registros em áudio de 16 reuniões de duas equipes de saúde da família, contando com 26 participantes, dentre eles profissionais e estagiários de diferentes especialidades. A análise foi feita a partir da perspectiva das práticas discursivas, com os seguintes passos: 1) transcrição do material; 2) leitura intensiva e organização do material; 3) construção de sentidos sobre a família, e análise dos repertórios interpretativos, discursos usados, e implicações para ação; e 4) narrativa ilustrativa da dinâmica dos sentidos. Os sentidos construídos na análise foram: a) Família como pessoas que moram juntas: os repertórios usados descreviam os modos de ser família a partir do ambiente em que ela vive, entendendo a família como informante e cuidadora dos seus membros; b) Família como responsável pelo cuidado: repertórios de família como aquela que dá suporte aos seus membros e é responsável por eles; e, por vezes, está sobrecarregada com esses cuidados; c) Família como problema: repertórios que configuravam a família como aquela que é responsável pelo problema de saúde dos seus membros, como aquela que funciona como um estressor para eles ou como aquela que está em situação de risco; e d) Família como rede de relações, sentido que foi usado, mais comumente, em conversas sobre casos complexos, com discussões voltadas para configurações, estruturas e dinâmicas familiares. A partir da análise do processo de discussão da equipe em torno de um \"caso\", foi possível ilustrar o dinamismo desses sentidos nas conversas e como esses são negociados a todo momento. A análise nos permite considerar que há esforços dos profissionais em voltar a atenção do cuidado para a família, porém ainda são comuns práticas centradas no indivíduo e pouco pautadas no contexto e nas condições de vida das famílias. Compreendemos que dar visibilidade a esses diferentes sentidos e seu uso permite reflexões sobre como cada forma de descrever as famílias possibilita a construção de práticas distintas, o que pode contribuir para uma maior reflexão dos profissionais de saúde sobre sua prática cotidiana (Apoio Capes e Fapesp- 2014/08618-6).

Acceptance and Commitment Therapy with Forensic Inpatients

This paper provides a preliminary exploration of the application of Acceptance and Commitment Therapy (ACT) within the context of a forensic hospital. ACT has a reputation for being a clinically flexible and empirically sound therapeutic intervention, which appears uniquely suited for forensic hospital settings. However, no research has been published to date on the use of ACT as a treatment for forensic inpatients. The ACT approach directly aims to help people let go of the unwinnable struggles to control symptoms of mental illness, and instead focus on constructing a "life worth living." ACT interventions can equip forensic patients with the values and flexible behavioral repertoires necessary to lead lives that are personally meaningful and satisfying and do not involve inflicting harm to others. The ACT model also attempts to minimize the therapist-patient hierarchy through an emphasis on the ubiquitous nature of human suffering. This approach can be particularly useful when working with marginalized, treatment-resistant patients. Continued research on the application of ACT with forensic inpatients is recommended.

El discurso profesional en servicios sociales comunitarios: una aproximación desde la identificación de repertorios interpretativos

Introducción: la presente investigación está orientada a ofrecer un análisis donde se establezcan los recursos lingüísticos utilizados por los participantes sobre el contenido y alcance de la prestación básica de información y orientación en los servicios sociales comunitarios, tal como la desarrollan los trabajadores sociales. Material y métodos: siguiendo una metodología cualitativa y la utilización del análisis del discurso en la propuesta de Wetherell y Potter (1996) con el empleo de la herramienta analítica de los repertorios interpretativos, se intentarán resaltar aquellos elementos definitorios, estrategias profesionales, valores, normas, prácticas organizacionales, elementos de la cultura institucional, entre otros, que dan forma a los escenarios donde desarrollan su labor los profesionales y que configuran el sistema de servicios sociales comunitarios. Resultados: las entrevistas realizadas a veinticinco trabajadores sociales de la provincia de Málaga muestran cuatro repertorios interpretativos que reflejan la construcción del sistema de servicios sociales por parte de los profesionales implicados: el olvido de lo comunitario, la eterna indefinición del sistema, el elefante encadenado y la escasez agudiza el ingenio. Discusión: se pone de manifiesto cómo se construye un modelo de intervención distante a lo establecido en las normas y códigos éticos a causa de los comportamientos organizacionales e institucionales, que los profesionales intentan minimizar mediante la puesta en práctica de habilidades personales.

TENDENCIA THATCHERITIS OR ENGLISHNESS REMADE: The Fictions of Julian Barnes, Hanif Kureishi and Pat Barker

Julian Barnes, Pat Barker, and Hanif Kureishi are all canonical authors whose fictions are widely believed to reflect the cultural and political state of a nation that is post-war, post-imperial and post-modern. While much has been written on how Barker’s and Kureishi’s early works in particular respond to and intervene in the presiding political narrative of the 1980s – Thatcherism – treatment of how revenants of Thatcherism have shaped these writers’ works from 1990 on has remained cursory. Thatcherism is more than an obvious historical reference point for Barker, Barnes, and Kureishi; their works demonstrate a sophisticated understanding of how Thatcher’s reworkings of the repertoires of Englishness – a representational as well as political and cultural endeavour – persist beyond her time in office. Barnes, Barker, and Kureishi seem to have reached the same conclusion as political and cultural critics: Thatcher and Thatcherism have remade not only the contemporary political and cultural landscapes but also the electorate and consequently the English themselves. Tony Blair’s conception of the New Britain proved less than satisfactory because contemporary repertoires of Englishness repeat and rework historical and not incidentally imperial formulations of England and Englishness rather than envision civic and populist formulations of renewal. Barnes’s England, England and Arthur & George confront the discourse of inevitability that has come to be attached to contemporary formulations of both political and cultural Englishness – both in terms of its predictable demise and its belated celebration. Kureishi’s The Buddha of Suburbia and “The Body” speak to an alteration that has taken place in which historical Englishness and Thatcherism have become complementary rather than contrasting discourses. What Barker’s Border Crossing and Double Vision offer against this backdrop is a subtle interrogation of how renewal itself comes to be a presiding mode of cultural reflection that absorbs revolutionary possibility.

Valuable recipes.

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Staging Theater to Realize a Nation: The Development of German National Theater in the 18th Century

Thesis (Ph.D.)--University of Washington, 2016-03

Locomotion and gaits of the northern brown bandicoot, Isoodon macrourus (Marsupalia: Peramelidae)

Gait repertoires of the northern brown bandicoot, Isoodon macrourus, were studied over a wide range of locomotor speeds. At low relative speeds, bandicoots used symmetrical gaits that included pacing, trotting, and lateral sequence strides. Forefoot contact duration was generally shorter than hind foot contact duration at all speeds. At moderate relative speeds bandicoots used half-bounding gaits with either no period of suspension or with a short gathered suspension. At high speeds the predominant gait had both a short extended and a short gathered suspension, although some strides comprised only an extended suspension. Increases in speed were accompanied by increases in spinal extension, presumably leading to the extended suspensions. On a stationary treadmill individuals occasionally used a bipedal gait. Maximum half-bounding speeds appear to be relatively low in this species.

Genes for the majority of group A streptococcal virulence factors and extracellular surface proteins do not confer an increased propensity to cause invasive disease

Background. The factors behind the reemergence of severe, invasive group A streptococcal (GAS) diseases are unclear, but it could be caused by altered genetic endowment in these organisms. However, data from previous studies assessing the association between single genetic factors and invasive disease are often conflicting, suggesting that other, as-yet unidentified factors are necessary for the development of this class of disease. Methods. In this study, we used a targeted GAS virulence microarray containing 226 GAS genes to determine the virulence gene repertoires of 68 GAS isolates (42 associated with invasive disease and 28 associated with noninvasive disease) collected in a defined geographic location during a contiguous time period. We then employed 3 advanced machine learning methods (genetic algorithm neural network, support vector machines, and classification trees) to identify genes with an increased association with invasive disease. Results. Virulence gene profiles of individual GAS isolates varied extensively among these geographically and temporally related strains. Using genetic algorithm neural network analysis, we identified 3 genes with a marginal overrepresentation in invasive disease isolates. Significantly, 2 of these genes, ssa and mf4, encoded superantigens but were only present in a restricted set of GAS M-types. The third gene, spa, was found in variable distributions in all M-types in the study. Conclusions. Our comprehensive analysis of GAS virulence profiles provides strong evidence for the incongruent relationships among any of the 226 genes represented on the array and the overall propensity of GAS to cause invasive disease, underscoring the pathogenic complexity of these diseases, as well as the importance of multiple bacteria and/ or host factors.

A DESMUSICALIZAÇÃO DA MÍDIA EM TEMPOS DE HIPERMIDIATIZAÇÃO DA MÚSICA: o empobrecimento estético da música popular na programação televisiva brasileira

O trabalho aborda o empobrecimento estético-discursivo da canção popular massiva exposta na mídia televisiva brasileira, na comparação entre o contexto histórico-midiático-musical das décadas de 1960 e 1970 e o contexto da primeira metade da segunda década do século XXI. Para definir tal empobrecimento discursivo encontrou-se o conceito de desmusicalização da mídia televisiva . O trabalho, de natureza teórica, tem como objetivo apontar que a mídia televisiva tem papel preponderante e parte importante da responsabilidade pelo empobrecimento na construção do discurso da música popular brasileira. Para tanto, primeiramente, por meio da análise poético-estética dos elementos estruturais das linguagens musical e literária de uma canção popular, delimitou-se a espécie de obra musical estruturada com acuro poético, que a ela empresta um padrão estético de excelência. Tais canções tinham espaços de veiculação nas grades de programação da mídia televisiva de outrora e nos dias atuais não mais os têm. Em seguida, por meio de pesquisas bibliográfica, documental e empírica, descreve-se de modo analítico-interpretativo o período histórico em que ocorreu essa desmusicalização da mídia. Por fim, constata-se que a partir da popularização da Internet e da proliferação de diversas mídias que permitem ao ouvinte o acesso, o armazenamento, o compartilhamento de repertórios musicais e, sobretudo, a audição em movimento, a canção popular massiva pautada pelo acuro poético deixou de estar presente na mídia televisiva e passou a figurar em outras diversas mídias. Essa ampla presença da Música nas mídias digitais definiu-se pelo conceito de hipermidiatização da música, que seria um dos componentes da desmusicalização da mídia televisiva.

Collective action and civil disobedience: the anti-GMO campaign of the Faucheurs volontaires

Civil disobedience has hitherto enjoyed only a relatively marginal place in the repertoires of French social movements, but has recently emerged as a key rallying frame for social mobilization, especially among environmental and counter-globalization movements. This paper examines the theory and practice of civil disobedience in the French context through an analysis of one such movement, the anti-GM Faucheurs Volontaires. Discussing the highly controversial campaign's positioning as 'civic disobedience', the article examines contested discourses of violence surrounding crop destruction, and the state responses to action, before asking what the campaign's claims to Republican civism mean for traditional notions of the relationship between state and challenging groups in France. It argues that framing action as civil disobedience is central to attempts to construct political and popular legitimacy, in terms of the campaign's national, international, and sectoral goals.

"Well did you feel jealous?" Control and ideology in police interviews with rape complainants

The methods used by the UK Police to investigate complaints of rape have unsurprisingly come under much scrutiny in recent times, with a 2007 joint report on behalf of HM Crown Prosecution Service Inspectorate and HM Inspectorate of Constabulary concluding that there were many areas where improvements should be made. The research reported here forms part of a larger project which draws on various discourse analytical tools to identify the processes at work during police interviews with women reporting rape. Drawing on a corpus of video recorded police interviews with women reporting rape, this study applies a two pronged analysis to reveal the presence of these ideologies. Firstly, an analysis of the discourse markers ‘well’ and ‘so’ demonstrates the control exerted on the interaction by interviewing officers, as they attach importance to certain facts while omitting much of the information provided by the victim. Secondly, the interpretative repertoires relied upon by officers to ‘make sense’ of victim’s accounts are subject to scrutiny. As well as providing micro-level analyses which demonstrate processes of interactional control at the local level, the findings of these analyses can be shown to relate to a wider context – specifically prevailing ideologies about sexual violence in society as a whole.

Computer based monitoring in U.K. service organizations : a comparative study

The majority of the literature about CBM is American in origin, and (inter alia) notes that there were differing uses of similar technology, indicating that context has an important role to play in the use of CBM. The literature maps the psychological effects of CBM in considerable detail, but only two published studies examine the context of CBM. These grounded results provide scant support for any systematic, quantitative, large scale analysis of computer based monitoring in the UK context. This thesis thus aims to systemically examine the context of CBM using discourse analysis. Forty four interviewees were theoretically sampled using a structured sample technique in four organizations. All were national or multinational enterprises. The interviews were semi structured in nature and divided into three sections. The first addressed the respondents' thoughts and perceptions about CBM, the second elicited talk about the departmental context (focusing the management - worker relationship), and the final section addressed the organizational context. The cases demonstrated variation in the use of CBM, measured according to the criteria of Westin (1987, 1988) and according to the interpretive repertoires used by the respondents in each case. Seven analytical categories of talk emerged from the data: three at the organizational level and four at the departmental level of analysis. Discourse analysis revealed two discrete interpretive repertories - the procedural and the substantive repertoires - in respondents' talk whose main variation occurred at the departmental level of analysis. Furthermore, patterns were found in the use of these repertories within cases and between categories. Between the cases, variation in the use of the repertories matched the between case variation according to the criteria of Westin. It would thus appear that the source of variation in the use of CBM lies in its context, more specifically in the relative emphasis of humanistic, interpersonal and idiosyncratic values within the management worker relationship.

The role of neighborhood organizations in the production of gentrifiable urban space: The case of Wynwood, Miami's Puerto Rican barrio

Partnerships between government and community-based actors and organizations are considered the hallmark of contemporary governance arrangements for the revitalization and gentrification of economically distressed, inner city areas. This dissertation uses historical, narrative analysis and ethnographic methods to examine the formation, evolution and operation of community-based governance partnerships in the production of gentrifiable urban space in the Wynwood neighborhood of Miami, FL between 1970 and 2010. This research is based on more than four years of participant observation, 60 in-depth interviews with respondents recruited through a purposive snowball sample, review of secondary and archival sources, and descriptive, statistical and GIS analysis. This study examines how different organizations formed in the neighborhood since the 1970s have facilitated the recent gentrification of Wynwood. It reveals specifically how partnerships between neighborhood-based government agencies, nonprofit organizations and real estate developers were constructed to be exclusionary and lead to inequitable economic development outcomes for Wynwood residents. The key factors conditioning these inequalities include both the rationalities of action of the organizations involved and the historical contexts in which their leaders’ thinking and actions were shaped. The historical contexts included the ethnic politics of organizational funding in the 1970s and the “entrepreneurial” turn of community-based economic development and Miami urban politics since the 1980s. Over time neighborhood organizations adopted highly pragmatic rationalities and repertoires of action. By the 2000s when Wynwood experienced unprecedented investment and redevelopment, the pragmatism of community-based organizations led them to become junior partners in governance arrangements and neighborhood activists were unable to directly challenge the inequitable processes and outcomes of gentrification.

Roles of CTCF and YY1 in T Cell Receptor Gene Rearrangement And T Cell Development

Diversity of T cell receptors (TCR) and immunoglobulins (Ig) is generated by V(D)J recombination of antigen receptor (AgR) loci. The Tcra-Tcrd locus is of particular interest because it displays a nested organization of Tcrd and Tcra gene segments and V(D)J recombination follows an intricate developmental program to assemble both TCRδ and TCRα repertoires. However, the mechanisms that dictate the developmental regulation of V(D)J recombination of the Tcra-Tcrd locus remain unclear.

We have previously shown that CCCTC-binding factor (CTCF) regulates Tcra gene transcription and rearrangement through organizing chromatin looping between CTCF- binding elements (CBEs). This study is one of many showing that CTCF functions as a chromatin organizer and transcriptional regulator genome-wide. However, detailed understanding of the impact of specific CBEs is needed to fully comprehend the biological function of CTCF and how CTCF influences the generation of the TCR repertoire during thymocyte development. Thus, we generated several mouse models with genetically modified CBEs to gain insight into the CTCF-dependent regulation of the Tcra-Tcrd locus. We revealed a CTCF-dependent chromatin interaction network at the Tcra-Tcrd locus in double-negative thymocytes. Disruption of a discrete chromatin loop encompassing Dδ, Jδ and Cδ gene segments allowed a single Vδ segment to frequently contact and rearrange to diversity and joining gene segments and dominate the adult TCRδ repertoire. Disruption of this loop also narrowed the TCRα repertoire, which, we believe, followed as a consequence of the restricted TCRδ repertoire. Hence, a single CTCF-mediated chromatin loop directly regulates TCRδ diversity and indirectly regulates TCRα diversity. In addition, we showed that insertion of an ectopic CBE can modify chromatin interactions and disrupt the rearrangement of particular Vδ gene segments. Finally, we investigated the role of YY1 in early T cell development by conditionally deleting YY1 in developing thymocytes. We found that early ablation of YY1 caused severe developmental defects in the DN compartment due to a dramatic increase in DN thymocyte apoptosis. Furthermore, late ablation of YY1 resulted in increased apoptosis of DP thymocytes and a restricted TCRα repertoire. Mechanistically, we showed that p53 was upregulated in both DN and DP YY1-deficient thymocytes. Eliminating p53 in YY1-deficient thymocytes rescued the survival and developmental defects, indicating that these YY1-dependent defects were p53-mediated. We conclude that YY1 is required to maintain cell viability during thymocyte development by thwarting the accumulation of p53.

Overall, this thesis work has shown that CTCF-dependent looping provides a central framework for lineage- and developmental stage-specific regulation of Tcra-Tcrd gene expression and rearrangements. In addition, we identified YY1 as a novel regulator of thymocyte viability.