Recombinant expression of Munc18c in a baculovirus system and interaction with syntaxin4

Two protein families that are critical for vesicle transport are the Syntaxin and Munc18/Sec1. families of proteins. These two molecules form a high affinity complex and play an essential role in vesicle docking and fusion. Munc18c was expressed as an N-terminally His-tagged fusion protein from recombinant baculovirus in Sf9 insect cells. His-tagged Munc18c was purified to homogeneity using both cobalt-chelating affinity chromatography and gel filtration chromatography. With this simple two-step protocol, 3.5 mg of purified Munc18c was obtained from a 1 L culture. Further, the N-terminal His-tag could be removed by thrombin cleavage while the tagged protein was bound to metal affinity resin. Recombinant Munc18c produced in this way is functional, in that it forms a stable complex with the SNARE interacting partner, syntaxin4. Thus we have developed a method for producing and purifying large amounts of functional Munc18c-both tagged and detagged-from a baculovirus expression system. We have also developed a method to purify the Munc18c:syntaxin4 complex. These methods will be employed for future functional and structural studies. Crown copyright (C) 2003 Published by Elsevier Inc. All rights reserved.

Endogenous angiotensin II modulates nNOS expression in renovascular hypertension

Nitric oxide (NO) influences renal blood flow mainly as a result of neuronal nitric oxide synthase (nNOS). Nevertheless, it is unclear how nNOS expression is modulated by endogenous angiotensin II, an inhibitor of NO function. We tested the hypothesis that the angiotensin II AT1 receptor and oxidative stress mediated by NADPH oxidase contribute to the modulation of renal nNOS expression in two-kidney, one-clip (2K1C) hypertensive rats. Experiments were performed on male Wistar rats (150 to 170 g body weight) divided into 2K1C (N = 19) and sham-operated (N = 19) groups. nNOS expression in kidneys of 2K1C hypertensive rats (N = 9) was compared by Western blotting to that of 2K1C rats treated with low doses of the AT1 antagonist losartan (10 mg·kg-1·day-1; N = 5) or the superoxide scavenger tempol (0.2 mmol·kg-1·day-1; N = 5), which still remain hypertensive. After 28 days, nNOS expression was significantly increased by 1.7-fold in the clipped kidneys of 2K1C rats and by 3-fold in the non-clipped kidneys of 2K1C rats compared with sham rats, but was normalized by losartan. With tempol treatment, nNOS expression increased 2-fold in the clipped kidneys and 1.4-fold in the non-clipped kidneys compared with sham rats. The changes in nNOS expression were not followed by changes in the enzyme activity, as measured indirectly by the cGMP method. In conclusion, AT1 receptors and oxidative stress seem to be primary stimuli for increased nNOS expression, but this up-regulation does not result in higher enzyme activity.

Design gráfico : análise da relação com a performance da micro e pequena empresa

O design possui um papel central nos negócios, participando de forma crucial no intercambio cultural e econômico da sociedade. Uma das competências do profissional de design gráfico é o desenvolvimento da Identidade Visual Corporativa (IVC). Ela tem como função definir visualmente o perfil de uma empresa, configurando-se, assim, como um composto mercadológico de fundamental importância para diferenciação das organizações, possuindo um importante papel para o crescimento das micro e pequenas empresas (MPEs). O objetivo desta pesquisa é verificar de que forma os gestores de MPEs percebem a atuação do design gráfico sobre a performance do seu negócio, avaliada a partir da sua experiência com a IVC desenvolvida por um profissional de design e aplicada pela empresa em suas atividades. A pesquisa é aplicada, em relação a sua finalidade, e exploratória, quanto ao seu objetivo. Sua condução se deu a partir de pesquisa bibliográfica, entrevista semiestruturada com os gestores das MPEs investigadas e pesquisa documental em materiais fornecidos pelos sujeitos da pesquisa, realizando-se, desta forma, uma triangulação metodológica a fim de contribuir para o exame do fenômeno. Como estratégia de investigação foi utilizado o estudo de casos múltiplos, realizado com 7 (sete) MPEs que haviam incorporado sua IVC há pelo menos 2 (dois) anos, dos setores de comércio e serviço, localizadas nos municípios de Vitória, Vila Velha, Serra ou Cariacica. Os dados coletados foram analisados por meio da Análise de Conteúdo, com base nas etapas descritas por Bardin (1977) e Laville (1999), distribuindo as unidades de análise (frases e parágrafos) nas categorias desenvolvidas: Motivação, Integração da IVC, Gestão da IVC, Relevância da IVC, IVC e Performance e Expressão Visual. A análise utilizou a abordagem quantitativa, realizada por meio da técnica de percentagem, e qualitativa, com maior ênfase na última, onde foram analisadas as categorias e seus elementos, assim como as relações entre elas, buscando extrair os significados construídos. Desta forma, foi possível verificar que a maior parte dos gestores das MPEs investigadas identificam que o design gráfico, por meio da IVC, contribui de forma positiva para a performance do seu negócio, proporcionando diferentes tipos de benefícios, dentre os mais citados foram: Identificação/Reconhecimento, Fortalecimento (solidez, estabilidade, profissionalismo), Novos Clientes, Imagem, Receptividade, Agregação de Valor para a Marca e Diferenciação.

Design gráfico : análise da relação com a performance da micro e pequena empresa

O design possui um papel central nos negócios, participando de forma crucial no intercambio cultural e econômico da sociedade. Uma das competências do profissional de design gráfico é o desenvolvimento da Identidade Visual Corporativa (IVC). Ela tem como função definir visualmente o perfil de uma empresa, configurando-se, assim, como um composto mercadológico de fundamental importância para diferenciação das organizações, possuindo um importante papel para o crescimento das micro e pequenas empresas (MPEs). O objetivo desta pesquisa é verificar de que forma os gestores de MPEs percebem a atuação do design gráfico sobre a performance do seu negócio, avaliada a partir da sua experiência com a IVC desenvolvida por um profissional de design e aplicada pela empresa em suas atividades. A pesquisa é aplicada, em relação a sua finalidade, e exploratória, quanto ao seu objetivo. Sua condução se deu a partir de pesquisa bibliográfica, entrevista semiestruturada com os gestores das MPEs investigadas e pesquisa documental em materiais fornecidos pelos sujeitos da pesquisa, realizando-se, desta forma, uma triangulação metodológica a fim de contribuir para o exame do fenômeno. Como estratégia de investigação foi utilizado o estudo de casos múltiplos, realizado com 7 (sete) MPEs que haviam incorporado sua IVC há pelo menos 2 (dois) anos, dos setores de comércio e serviço, localizadas nos municípios de Vitória, Vila Velha, Serra ou Cariacica. Os dados coletados foram analisados por meio da Análise de Conteúdo, com base nas etapas descritas por Bardin (1977) e Laville (1999), distribuindo as unidades de análise (frases e parágrafos) nas categorias desenvolvidas: Motivação, Integração da IVC, Gestão da IVC, Relevância da IVC, IVC e Performance e Expressão Visual. A análise utilizou a abordagem quantitativa, realizada por meio da técnica de percentagem, e qualitativa, com maior ênfase na última, onde foram analisadas as categorias e seus elementos, assim como as relações entre elas, buscando extrair os significados construídos. Desta forma, foi possível verificar que a maior parte dos gestores das MPEs investigadas identificam que o design gráfico, por meio da IVC, contribui de forma positiva para a performance do seu negócio, proporcionando diferentes tipos de benefícios, dentre os mais citados foram: Identificação/Reconhecimento, Fortalecimento (solidez, estabilidade, profissionalismo), Novos Clientes, Imagem, Receptividade, Agregação de Valor para a Marca e Diferenciação.

Identity and the concept of the West: the case of Brazil and India

This study aims to provide insights about how Brazil and India view and relate to the concept of the West, and how this affects their identity and foreign policy. Both countries' notions about the West are the subject of lively domestic discussion both in academia and the media, reflecting the struggle these countries find themselves in to define their identity as they rise. I argue that the concept of the West serves, in both Brazil and India, as a crucial concept to articulate their own identity-by a complex combination of criticizing, distancing itself from, or attempting to emulate the West.

Teaching and learning production and operations management: the journey from identity crisis to a cross, disciplinary approach

Este artigo tem suas raízes em algumas questões relacionadas à "forma" e ao "conteúdo" do que nós, professores, ensinamos na área de Administração da Produção e Operações. Inicialmente, descrevo a evolução histórica desse campo no Brasil. Em seguida, discuto a crise de identidade que o campo está sofrendo. Com o objetivo de apresentar respostas para essa situação, apresento seis propostas para o desenvolvimento e consolidação do campo. Finalmente, descrevo uma iniciativa prática, envolvendo uma disciplina específica da área, ensinada para alunos de pós-graduação. Essa iniciativa enfatiza a "dimensão do conteúdo" (de uma abordagem técnico-operacional para uma abordagem estratégico-gerencial) como também a "dimensão da forma" (do foco no ensino para o foco no aprendizado). O sucesso dessa experiência em curso confirma a coerência da agenda proposta e induz futuros aperfeiçoamentos.

Territorial Identity and Sustainable Development: from Concept to Analysis

A necessidade de (re)afirmar a identidade dos lugares e regiões com o objectivo de aumentar a sua competitividade no mercado global de bens, serviços e ideias tem sido geralmente adoptado nas políticas de desenvolvimento regional e local, mais de um modo retórico do que operacional. Faltam, de facto, instrumentos analíticos adequados para a avaliação da identidade territorial no contexto do nexo local-global. Neste artigo é discutido o enquadramento conceptual e metodológico necessário para o estudo das mudanças nas identidades territoriais e é apresentada evidência empírica dos conhecimentos, atitudes e práticas dos agentes de desenvolvimento local em Portugal.

Is there One Goan Identity, Several or None?

A identidade goesa: uma, várias ou nenhuma? A lingua materna e as experiências históricas da comunidade ancestral são componentes essenciais desse legado. Em caso dos goeses que vivem em Goa ou tiveram antepassados goeses, a matriz cultural indiana está sempre presente, mesmo que seja em forma mais ou menos diluida. A presença colonial portuguesa deixou a sua marca, tanto nos goeses católicos, como nos goeses hindus. Sem esta componente dificilmente Goa poderia justificar o direito para o estatuto de Estado autónomo na união indiana. E como é com os goeses em Portugal? A consciencia importada das castas continua a jogar o seu papel nas várias associações em que predomina uma casta ou outra. Mas nada disso impede os goeses em Portugal de apreciar a carilada ou os cantares tradicionais de Goa. Nem perdem alguma oportunidade de visitar a sua terra natal.

Unwrapping Goan Identity

O título do ensaio evoca Gunter Grass. É um apelo aos goeses para se reconciliarem com o seu legado histórico. O autor revela as suas dificuldades nesse sentido até se sentir confortável com a multiplicidade das experiência que são consideradas como um enriquecimento da identidade. Refere ao desafio que continua a atenuar as capacidades dos goeses em globalização: o legado das castas.

Eastwards / westwards: which direction for gender studies in the 21st century?

Eastwards / Westwards: Which Direction for Gender Studies in the XXIst Century? is a collection of essays which focus on themes and methods that characterize current research into gender in Asian countries in general. In this collection, ideas derived from Gender Studies elsewhere in the world have been subjected to scrutiny for their utility in helping to describe and understand regional phenomena. But the concepts of Local and Global – with their discoursive productions – have not functioned as a binary opposition: localism and globalism are mutually constitutive and researchers have interrogated those spaces of interaction between the ‘self’ and the ‘other’, bearing in mind their own embeddedness in social and cultural structures and their own historical memory. Contributors to this collection provided a critical transnational perspective on some of the complex effects of the dynamics of cultural globalization, by exploring the relation between gender and development, language, historiography, education and culture. We have also given attention to the ideological and rhetorical processes through which gender identity is constructed, by comparing textual grids and patterns of expectation. Likewise, we have discussed the role of ethnography, anthropology, historiography, sociology, fiction, popular culture and colonial and post-colonial sources in (re)inventing old/new male/female identities, their conversion into concepts and circulation through time and space. This multicultural and trans-disciplinary selection of essays is totally written in English, fully edited and revised, therefore, it has a good potential for an immediate international circulation. This project may trace new paths and issues for discussion on what concerns the life, practices and narratives by and about women in Asia, as well as elsewhere in the present day global experience. Academic readership: Researchers, scholars, educators, graduate and post-graduate students, doctoral students and general non-fiction readers, with a special interest in Gender Studies, Asia, Colonial and Post-Colonial Literature, Anthropology, Cultural Studies, History, Historiography, Politics, Race, Feminism, Language, Linguistics, Power, Political and Feminist Agendas, Popular Culture, Education, Women’s Writing, Religion, Multiculturalism, Globalisation, Migration. Chapter summary: 1. “Social Gender Stereotypes and their Implication in Hindi”, Anjali Pande, Jawaharlal Nehru University, New Delhi, India. This essay looks at the subtle ways in which gender identities are constructed and reinforced in India through social norms of language use. Language itself becomes a medium for perpetuating gender stereotypes, forcing its speakers to confirm to socially defined gender roles. Using examples from a classroom discussion about a film, this essay will highlight the underlying rigid male-female stereotypes in Indian society with their more obvious expressions in language. For the urban woman in India globalisation meant increased economic equality and exposure to changed lifestyles. On an individual level it also meant redefining gender relations and changing the hierarchy in man-­woman relationships. With the economic independence there is a heightened sense of liberation in all spheres of social life, a confidence to fuzz the rigid boundaries of gender roles. With the new films and media celebrating this liberated woman, who is ready to assert her sexual needs, who is ready to explode those long held notions of morality, one would expect that the changes are not just superficial. But as it soon became obvious in the course of a classroom discussion about relationships and stereotypes related to age, the surface changes can not become part of the common vocabulary, for the obvious reason that there is still a vast gap between the screen image of this new woman and the ground reality. Social considerations define the limits of this assertiveness of women, whereas men are happy to be liberal within the larger frame of social sanctions. The educated urban woman in India speaks in favour of change and the educated urban male supports her, but one just needs to scratch the surface to see the time tested formulae of gender roles firmly in place. The way the urban woman happily balances this emerging promise of independence with her gendered social identity, makes it necessary to rethink some aspects of looking at gender in a gradually changing, traditional society like India. 2. “The Linguistic Dimension of Gender Equality”, Alissa Tolstokorova, Kiev Centre for Gender Information and Education, Ukraine. The subject-matter of this essay is gender justice in language which, as I argue, may be achieved through the development of a gender-related approach to linguistic human rights. The last decades of the 20th century, globally marked by a “gender shift” in attitudes to language policy, gave impetus to the social movement for promoting linguistic gender equality. It was initiated in Western Europe and nowadays is moving eastwards, as ideas of gender democracy progress into developing countries. But, while in western societies gender discrimination through language, or linguistic sexism, was an issue of concern for over three decades, in developing countries efforts to promote gender justice in language are only in their infancy. My argument is that to promote gender justice in language internationally it is necessary to acknowledge the rights of women and men to equal representation of their gender in language and speech and, therefore, raise a question of linguistic rights of the sexes. My understanding is that the adoption of the Universal Declaration of Linguistic Rights in 1996 provided this opportunity to address the problem of gender justice in language as a human rights issue, specifically as a gender dimension of linguistic human rights. 3. “The Rebirth of an Old Language: Issues of Gender Equality in Kazakhstan”, Maria Helena Guimarães, Polytechnic Institute of Porto, Portugal. The existing language situation in Kazakhstan, while peaceful, is not without some tension. We propose to analyze here some questions we consider relevant in the frame of cultural globalization and gender equality, such as: free from Russian imperialism, could Kazakhstan become an easy prey of Turkey’s “imperialist dream”? Could these traditionally Muslim people be soon facing the end of religious tolerance and gender equality, becoming this new old language an easy instrument for the infiltration in the country of fundamentalism (it has already crossed the boarders of Uzbekistan), leading to a gradual deterioration of its rich multicultural relations? The present structure of the language is still very fragile: there are three main dialects and many academics defend the re-introduction of the Latin alphabet, thus enlarging the possibility of cultural “contamination” by making the transmission of fundamentalist ideas still easier through neighbour countries like Azerbaijan, Uzbekistan and Turkmenistan (their languages belong to the same sub-group of Common Turkic), where the Latin alphabet is already in use, and where the ground for such ideas shown itself very fruitful. 4. “Construction of Womanhood in the Bengali Language of Bangladesh”, Raasheed Mahmood; University of New South Wales, Sydney. The present essay attempts to explore the role of gender-based language differences and of certain markers that reveal the status accorded to women in Bangladesh. Discrimination against women, in its various forms, is endemic in communities and countries around the world, cutting across class, race, age, and religious and national boundaries. One cannot understand the problems of gender discrimination solely by referring to the relationship of power or authority between men and women. Rather one needs to consider the problem by relating it to the specific social formation in which the image of masculinity and femininity is constructed and reconstructed. Following such line of reasoning this essay will examine the nature of gender bias in the Bengali language of Bangladesh, holding the conviction that as a product of social reality language reflects the socio-cultural behaviour of the community who speaks it. This essay will also attempt to shed some light on the processes through which gender based language differences produce actual consequences for women, who become exposed to low self-esteem, depression and systematic exclusion from public discourse. 5. “Marriage in China as an expression of a changing society”, Elisabetta Rosado David, University of Porto, Portugal, and Università Ca’Foscari, Venezia, Italy. In 29 April 2001, the new Marriage Law was promulgated in China. The first law on marriage was proclaimed in 1950 with the objective of freeing women from the feudal matrimonial system. With the second law, in 1981, values and conditions that had been distorted by the Cultural Revolution were recovered. Twenty years later, a new reform was started, intending to update marriage in the view of the social and cultural changes that occurred with Deng Xiaoping’s “open policy”. But the legal reform is only the starting point for this case-study. The rituals that are followed in the wedding ceremony are often hard to understand and very difficult to standardize, especially because China is a vast country, densely populated and characterized by several ethnic minorities. Two key words emerge from this issue: syncretism and continuity. On this basis, we can understand tradition in a better way, and analyse whether or not marriage, as every social manifestation, has evolved in harmony with Chinese culture. 6. “The Other Woman in the Portuguese Colonial Empire: The Case of Portuguese India”, Maria de Deus Manso, University of Évora, Portugal. This essay researches the social, cultural and symbolic history of local women in the Portuguese Indian colonial enclaves. The normative Portuguese overseas history has not paid any attention to the “indigenous” female populations in colonial Portuguese territories, albeit the large social importance of these social segments largely used in matrimonial and even catholic missionary strategies. The first attempt to open fresh windows in the history of this new field was the publication of Charles Boxer’s referential study about Women in lberian Overseas Expansion, edited in Portugal only after the Revolution of 1975. After this research we can only quote some other fragmentary efforts. In fact, research about the social, cultural, religious, political and symbolic situation of women in the Portuguese colonial territories, from the XVI to the XX century, is still a minor historiographic field. In this essay we discuss this problem and we study colonial representations of women in the Portuguese Indian enclaves, mainly in the territory of Goa, using case studies methodologies. 7. “Heading East this Time: Critical Readings on Gender in Southeast Asia”, Clara Sarmento, Polytechnic Institute of Porto, Portugal. This essay intends to discuss some critical readings of fictional and theoretical texts on gender condition in Southeast Asian countries. Nowadays, many texts about women in Southeast Asia apply concepts of power in unusual areas. Traditional forms of gender hegemony have been replaced by other powerful, if somewhat more covert, forms. We will discuss some universal values concerning conventional female roles as well as the strategies used to recognize women in political fields traditionally characterized by male dominance. Female empowerment will mean different things at different times in history, as a result of culture, local geography and individual circumstances. Empowerment needs to be perceived as an individual attitude, but it also has to be facilitated at the macro­level by society and the State. Gender is very much at the heart of all these dynamics, strongly related to specificities of historical, cultural, ethnic and class situatedness, requiring an interdisciplinary transnational approach.

Culture, politics and identity: critical readings on gender in southeast Asia

Indian Journal of Gender Studies October 2012 vol. 19 no. 3 437-467

The expression of tubulin cofactor A (TBCA) is regulated by a noncoding antisense Tbca RNA during testis maturation

Abstract - Recently, long noncoding RNAs have emerged as pivotal molecules for the regulation of coding genes' expression. These molecules might result from antisense transcription of functional genes originating natural antisense transcripts (NATs) or from transcriptional active pseudogenes. TBCA interacts with β-tubulin and is involved in the folding and dimerization of new tubulin heterodimers, the building blocks of microtubules. Methodology/Principal findings: We found that the mouse genome contains two structurally distinct Tbca genes located in chromosomes 13 (Tbca13) and 16 (Tbca16). Interestingly, the two Tbca genes albeit ubiquitously expressed, present differential expression during mouse testis maturation. In fact, as testis maturation progresses Tbca13 mRNA levels increase progressively, while Tbca16 mRNA levels decrease. This suggests a regulatory mechanism between the two genes and prompted us to investigate the presence of the two proteins. However, using tandem mass spectrometry we were unable to identify the TBCA16 protein in testis extracts even in those corresponding to the maturation step with the highest levels of Tbca16 transcripts. These puzzling results led us to re-analyze the expression of Tbca16. We then detected that Tbca16 transcription produces sense and natural antisense transcripts. Strikingly, the specific depletion by RNAi of these transcripts leads to an increase of Tbca13 transcript levels in a mouse spermatocyte cell line. Conclusions/Significance: Our results demonstrate that Tbca13 mRNA levels are post-transcriptionally regulated by the sense and natural antisense Tbca16 mRNA levels. We propose that this regulatory mechanism operates during spermatogenesis, a process that involves microtubule rearrangements, the assembly of specific microtubule structures and requires critical TBCA levels.