THE MECHANISM OF TUMORIGENESIS IN THE IMMORTALIZED HUMAN PANCREATIC CELL LINES: CELL CULTURE MODELS OF HUMAN PANCREATIC CANCER

The mechanism of tumorigenesis in the immortalized human pancreatic cell lines: cell culture models of human pancreatic cancer Pancreatic ductal adenocarcinoma (PDAC) is the most lethal cancer in the world. The most common genetic lesions identified in PDAC include activation of K-ras (90%) and Her2 (70%), loss of p16 (95%) and p14 (40%), inactivation p53 (50-75%) and Smad4 (55%). However, the role of these signature gene alterations in PDAC is still not well understood, especially, how these genetic lesions individually or in combination contribute mechanistically to human pancreatic oncogenesis is still elusive. Moreover, a cell culture transformation model with sequential accumulation of signature genetic alterations in human pancreatic ductal cells that resembles the multiple-step human pancreatic carcinogenesis is still not established. In the present study, through the stepwise introduction of the signature genetic alterations in PDAC into the HPV16-E6E7 immortalized human pancreatic duct epithelial (HPDE) cell line and the hTERT immortalized human pancreatic ductal HPNE cell line, we developed the novel experimental cell culture transformation models with the most frequent gene alterations in PDAC and further dissected the molecular mechanism of transformation. We demonstrated that the combination of activation of K-ras and Her2, inactivation of p16/p14 and Smad4, or K-ras mutation plus p16 inactivation, was sufficient for the tumorigenic transformation of HPDE or HPNE cells respectively. We found that these transformed cells exhibited enhanced cell proliferation, anchorage-independent growth in soft agar, and grew tumors with PDAC histopathological features in orthotopic mouse model. Molecular analysis showed that the activation of K-ras and Her2 downstream effector pathways –MAPK, RalA, FAK, together with upregulation of cyclins and c-myc were involved in the malignant transformation. We discovered that MDM2, BMP7 and Bmi-1 were overexpressed in the tumorigenic HPDE cells, and that Smad4 played important roles in regulation of BMP7 and Bmi-1 gene expression and the tumorigenic transformation of HPDE cells. IPA signaling pathway analysis of microarray data revealed that abnormal signaling pathways are involved in transformation. This study is the first complete transformation model of human pancreatic ductal cells with the most common gene alterations in PDAC. Altogether, these novel transformation models more closely recapitulate the human pancreatic carcinogenesis from the cell origin, gene lesion, and activation of specific signaling pathway and histopathological features.

Trade and Culture in International Law : Paths to (Re)Conciliation

The UNESCO Convention on cultural diversity marks a wilful separation between the issues of trade and culture on the international level. The present article explores this intensified institutional, policy- and decision-making disconnect and exposes its flaws and the considerable drawbacks it brings with it. These drawbacks, the article argues, become particularly pronounced in the digital media environment that has impacted upon both the conditions of trade with cultural products and services and upon the diversity of cultural expressions in local and global contexts. Criticising the strong and now increasingly meaningless path dependencies of the analogue age, the article sketches some possible ways to reconciling trade and culture, most of which lead back to the WTO, rather than to UNESCO.

Trade Versus Culture in the Digital Environment : An Old Conflict in Need of a New Definition

Following the recent UNESCO Convention on the Protection and Promotion of the Diversity of Cultural Expressions, the first wave of scholarly work has focused on clarifying the interface between the Convention and the WTO Agreements. Building upon these analyses, the present article takes however a different stance. It seeks a new, rather pragmatic definition of the relationship between trade and culture and argues that such a re-definition is particularly needed in the digital networked environment that has modified the ways markets for cultural content function and the ways in which cultural content is created, distributed and accessed. The article explores first the significance of the UNESCO Convention (or the lack thereof) and subsequently outlines a variety of ways in which the WTO framework can be improved in a ‘neutral’, not necessarily culturally motivated, manner to become more conducive to the pursuit of cultural diversity and taking into account the changed reality of digital media. The article also looks at other facets of the profoundly fragmented culture-related regulatory framework and underscores the critical importance of intellectual property rights and of other domains that appear at first sight peripheral to the trade and culture discussion, such as access to infrastructure, interoperability or net neutrality. It is argued that a number of feasible solutions exist beyond the politically charged confrontation of trade versus culture and that the new digital media landscape may require a readjustment of the priorities and the tools for the achievement of the widely accepted objective of cultural diversity.

Reconciling Trade and Culture : A Global Law Perspective

This article provides an overview of the most essential issues in the trade and culture discourse from a global law perspective. It looks into the intensified disconnect between trade and culture and exposes its flaws and the considerable drawbacks that it brings with it. It is argued that these drawbacks become especially pronounced in the digital media environment, which has strongly affected both the conditions of trade with cultural products and services and cultural diversity in local and global contexts. In this modified setting, there could have been a number of feasible ‘trade and culture’ solutions – i.e. regulatory designs that whilst enhancing trade liberalisation are also conducive to cultural policy. Yet, the realisation of any of these options becomes chimerical as the line between trade and culture matters is drawn in a clear and resolute manner. The article is meant for an interdisciplinary audience and forthcoming in the Journal of Arts Management, Law and Society.

Trade versus Culture : The Policy of Cultural Exception and the World Trade Organization

This chapter is a contribution to the Palgrave Handbook of European Media Policy (co-edited by Pauwels, Donders & Loisen). It is the chapter’s purpose to examine the proponents of the cultural exception policy, their strategies and demands, and to explore how they came to be reflected in the law and policy of the World Trade Organization (WTO). The chapter also looks at the current state of affairs, as although WTO law has not undergone any substantial amendments since its entry into force in 1995, the media landscape has in the meantime been truly transformed, in some aspects in a revolutionary manner. The broader picture of global governance has not remained still either, with new and emergent powers, changing mechanisms of rule-making and taking.

Transgressive sporting bodies - A theoretical and critical dicourse analysis of the representation of Caster Semenya in Swiss print media

A debate about Caster Semenya's female sex began shortly after the South African runner won gold in the women’s 800m final at the 2009 Athletic World Championships held in Berlin. Her victory was disputed through questions about her right to compete as a ‘woman’, with the International Association of Athletics Federation (IAAF) announcing she would be required to undergo a gender verification test before her victory could be confirmed. Using the theoretical frame of social constructionism (Berger & Luckmann), poststructuralism (Foucault), gender- and postcolonial theories (Butler; Hall; Spivak) and the methodology of critical discourse analysis (Jaeger), the paper explores the way the possible intersexuality of Caster Semenya was contextualised in mainstream Swiss German-language print media. The analyses will firstly look at the way in which Caster Semenya was constructed as a ʻfallen hero’ and stigmatised as a double-dealer and unacceptable deviant body. The rumours amongst athletes and commentators became news in the media, which focused on descriptions of her habitus, her muscular body and her deep voice. Through theoretical discussion the paper argues that the media response to Caster Semenya exemplifies Butler’s claim that the discursive framework of gender constructs and naturalises sex. A key question is therefore whether the designation of deviant bodies to a ʻfield of deformation’ (Butler) works to pluralise the field of gender, or rather, as Butler suggests, it tends that those bodies might call into questions. The final part of the paper discusses how gender, ethnicity and sexuality combine to constitute the black female sporting body as a spectacle of otherness. It is evident that this otherness is made manifest through the function of those bodies as a site of transgression, as the boundary between male and female, and often as the boundary between culture and nature (Hall). Using the example of the controversy surrounding Caster Semenya, this paper aims to demonstrate how the post/colonial white female body is reproduced by western norms of gender, sexuality, beauty and sporting behaviour, in the sense of a feminine sporting genderperformance. The media controversy will be also read through the lens of the globalisation of certain ideas of normative bodies, sex, ethnicity and gender and the challenge of changing stereotypes through transgression. Keywords: gender- and postcolonial theories, discourse analysis, print media, Caster Semen-ya, deviant body, ethnicity, intersexuality

Transgressive sporting bodies - A theoretical and critical dicourse analysis of the representation of Caster Semenya in Swiss print media

A debate about Caster Semenya's female sex began shortly after the South African runner won gold in the women’s 800m final at the 2009 Athletic World Championships held in Berlin. Her victory was disputed through questions about her right to compete as a ‘woman’, with the International Association of Athletics Federation (IAAF) announcing she would be required to undergo a gender verification test before her victory could be confirmed. Using the theoretical frame of social constructionism (Berger & Luckmann), poststructuralism (Foucault), gender- and postcolonial theories (Butler; Hall; Spivak) and the methodology of critical discourse analysis (Jaeger), the paper explores the way the possible intersexuality of Caster Semenya was contextualised in mainstream Swiss German-language print media. The analyses will firstly look at the way in which Caster Semenya was constructed as a ʻfallen hero’ and stigmatised as a double-dealer and unacceptable deviant body. The rumours amongst athletes and commentators became news in the media, which focused on descriptions of her habitus, her muscular body and her deep voice. Through theoretical discussion the paper argues that the media response to Caster Semenya exemplifies Butler’s claim that the discursive framework of gender constructs and naturalises sex. A key question is therefore whether the designation of deviant bodies to a ʻfield of deformation’ (Butler) works to pluralise the field of gender, or rather, as Butler suggests, it tends that those bodies might call into questions. The final part of the paper discusses how gender, ethnicity and sexuality combine to constitute the black female sporting body as a spectacle of otherness. It is evident that this otherness is made manifest through the function of those bodies as a site of transgression, as the boundary between male and female, and often as the boundary between culture and nature (Hall). Using the example of the controversy surrounding Caster Semenya, this paper aims to demonstrate how the post/colonial white female body is reproduced by western norms of gender, sexuality, beauty and sporting behaviour, in the sense of a feminine sporting genderperformance. The media controversy will be also read through the lens of the globalisation of certain ideas of normative bodies, sex, ethnicity and gender and the challenge of changing stereotypes through transgression. Keywords: gender- and postcolonial theories, discourse analysis, print media, Caster Semen-ya, deviant body, ethnicity, intersexuality

CCL5/RANTES is a key chemoattractant released by degenerative intervertebral discs in organ culture.

Release of chemotactic factors in response to tissue damage has been described for different musculoskeletal tissues, including the intervertebral disc (IVD). This study investigated the chemoattractants that are released by induced degenerative IVDs and may be involved in recruiting mesenchymal stem cells (MSCs). Bovine caudal discs were cultured within a bioreactor and loaded under conditions that mimicked physiological or degenerative settings. Between days 4-6, medium was replaced by PBS, which was subsequently used for proteomic, ELISA and immunoprecipitation analyses of secreted chemokines and cytokines. A Boyden chamber assay was used to observe human MSC migration towards native and chemokine depleted media. Gene expression levels of chemokine receptors in human MSCs were analysed, and CCL5 was localised in bovine and human IVD by immunohistochemistry. Proteomic analysis revealed the presence of CCL5 and CXCL6 within conditioned media. Higher concentrations of CCL5 were found in the degenerative media, and a relationship was found between interleukin-1β and CCL5 concentration. Chemokine immunoprecipitation showed that MSCs had a significantly reduced chemotactic migration towards CCL5-immunoprecipitated and CCL5/CXCL6 co-immunoprecipitated media, whilst CXCL6 depletion did not change MSC chemotaxis. MSCs showed a significant increase in mRNA expression of the CCL5 receptors, CCR1 and CCR4, upon culture in degenerative media. Furthermore, CCL5 was identified in bovine and human disc tissue by immunohistochemistry. Hence, CCL5 may be a key chemoattractant that is produced and released by the intervertebral disc cells. Therefore, these factors could be used to enhance stem/progenitor cell mobilisation in regenerative therapies for early stages of disc degeneration.

Time Course of Antibiotic and Antifungal Concentrations in Corneal Organ Culture.

PURPOSE Contamination with bacteria and/or fungi is a serious complication in organ-cultured corneas. Hence, antibiotic and antifungal agents are added to the culture medium. The concentration of different antimicrobial and antifungal additives to the media over time has so far not been investigated in detail and is the aim of this study. METHODS Nine human fresh corneoscleral discs were stored in corneal culture medium consisting of 2% fetal bovine serum and minimal essential medium. In addition, the culture medium contained 1200 μg/mL penicillin G, 25 μg/mL amphotericin B, 120 μg/mL streptomycin, and 100 μg/mL voriconazole. The concentration of amphotericin B used was 10 times higher than in clinical routine to facilitate its detection. The cultures were kept at 37°C for 28 days. At days 0, 7, 14, 21, and 28, samples of the culture medium were harvested for analysis of antimicrobial concentrations by liquid chromatography and electrospray ionization tandem mass spectrometry. RESULTS During corneal storage, the concentration of all antibiotics and antifungal agents declined significantly. By day 28, penicillin G was reduced to 14% of the original concentration. Amphotericin B and streptomycin retained approximately 60% of the original concentration to the end of the experiment and voriconazole maintained stable concentrations after an initial decline to approximately 80% at 7 days. CONCLUSIONS Throughout the entire storage period, the concentrations of penicillin G, streptomycin, and voriconazole exceeded the minimum inhibitory concentrations of all common contaminants, obviating the need for a change of the medium for antimicrobial reasons. Based on the minimum inhibitory concentrations and our findings, the initial concentration of amphotericin B should be raised to 5 μg/mL.

Textualidad oral - escrita - impresa en el pasaje Edad Media - Renacimiento

El objetivo de este trabajo es analizar el cambio de relaciones que se produce en Ia literatura española entre los ámbitos de Ia oralidad, Ia escritura manuscrita y la imprenta en el pasaje de la Edad Media al Renacimiento. A partir del estudio de estas diferentes "tecnologías de la palabra", citando el subtitulo del clásico libro de Walter Ong, se intenta esclarecer las características de un proceso que conduce a las manifestaciones literarias de Ia modernidad y esboza los primeros rasgos del fenómeno que más adelante se constituirá como Ia cultura de masas.

Esperando otro 17 de Octubre : La identidad de clase media y la experiencia de la crisis de 2001 en Argentina

Este trabajo se propone contribuir a una mejor comprensión de la dimensión política de la identidad de clase media en Argentina, a través del análisis de una imagen que apareció con cierta insistencia luego de 2001: la de un "17 de Octubre de la clase media". La curiosa imagen y los desplazamientos en sus sentidos, desde su primera aparición hasta sus usos más recientes, es interpretada reconduciéndola a dos contextos. Por una parte, el de los cambios socioeconómicos, políticos e identitarios que la sociedad argentina experimentó a partir de la década de 1990. Por la otra, el del trauma que la irrupción del peronismo produjo en 1945, cuyos efectos todavía persisten en la cultura nacional. La imagen del "17 de Octubre de la clase media" se interpreta así como una fantasía de superación del horizonte peronista por obra de un evento similar al que en 1945 protagonizaron los más pobres, pero esta vez encabezado por la clase media

Textualidad oral - escrita - impresa en el pasaje Edad Media - Renacimiento

El objetivo de este trabajo es analizar el cambio de relaciones que se produce en Ia literatura española entre los ámbitos de Ia oralidad, Ia escritura manuscrita y la imprenta en el pasaje de la Edad Media al Renacimiento. A partir del estudio de estas diferentes "tecnologías de la palabra", citando el subtitulo del clásico libro de Walter Ong, se intenta esclarecer las características de un proceso que conduce a las manifestaciones literarias de Ia modernidad y esboza los primeros rasgos del fenómeno que más adelante se constituirá como Ia cultura de masas.

Esperando otro 17 de Octubre : La identidad de clase media y la experiencia de la crisis de 2001 en Argentina

Este trabajo se propone contribuir a una mejor comprensión de la dimensión política de la identidad de clase media en Argentina, a través del análisis de una imagen que apareció con cierta insistencia luego de 2001: la de un "17 de Octubre de la clase media". La curiosa imagen y los desplazamientos en sus sentidos, desde su primera aparición hasta sus usos más recientes, es interpretada reconduciéndola a dos contextos. Por una parte, el de los cambios socioeconómicos, políticos e identitarios que la sociedad argentina experimentó a partir de la década de 1990. Por la otra, el del trauma que la irrupción del peronismo produjo en 1945, cuyos efectos todavía persisten en la cultura nacional. La imagen del "17 de Octubre de la clase media" se interpreta así como una fantasía de superación del horizonte peronista por obra de un evento similar al que en 1945 protagonizaron los más pobres, pero esta vez encabezado por la clase media

Textualidad oral - escrita - impresa en el pasaje Edad Media - Renacimiento

El objetivo de este trabajo es analizar el cambio de relaciones que se produce en Ia literatura española entre los ámbitos de Ia oralidad, Ia escritura manuscrita y la imprenta en el pasaje de la Edad Media al Renacimiento. A partir del estudio de estas diferentes "tecnologías de la palabra", citando el subtitulo del clásico libro de Walter Ong, se intenta esclarecer las características de un proceso que conduce a las manifestaciones literarias de Ia modernidad y esboza los primeros rasgos del fenómeno que más adelante se constituirá como Ia cultura de masas.

Esperando otro 17 de Octubre : La identidad de clase media y la experiencia de la crisis de 2001 en Argentina

Este trabajo se propone contribuir a una mejor comprensión de la dimensión política de la identidad de clase media en Argentina, a través del análisis de una imagen que apareció con cierta insistencia luego de 2001: la de un "17 de Octubre de la clase media". La curiosa imagen y los desplazamientos en sus sentidos, desde su primera aparición hasta sus usos más recientes, es interpretada reconduciéndola a dos contextos. Por una parte, el de los cambios socioeconómicos, políticos e identitarios que la sociedad argentina experimentó a partir de la década de 1990. Por la otra, el del trauma que la irrupción del peronismo produjo en 1945, cuyos efectos todavía persisten en la cultura nacional. La imagen del "17 de Octubre de la clase media" se interpreta así como una fantasía de superación del horizonte peronista por obra de un evento similar al que en 1945 protagonizaron los más pobres, pero esta vez encabezado por la clase media