The importance of connexin hemichannels during chondroprogenitor cell differentiation in hydrogel versus microtissue culture models

Appropriate selection of scaffold architecture is a key challenge in cartilage tissue engineering. Gap junction-mediated intercellular contacts play important roles in precartilage condensation of mesenchymal cells. However, scaffold architecture could potentially restrict cell-cell communication and differentiation. This is particularly important when choosing the appropriate culture platform as well as scaffold-based strategy for clinical translation, that is, hydrogel or microtissues, for investigating differentiation of chondroprogenitor cells in cartilage tissue engineering. We, therefore, studied the influence of gap junction-mediated cell-cell communication on chondrogenesis of bone marrow-derived mesenchymal stromal cells (BM-MSCs) and articular chondrocytes. Expanded human chondrocytes and BM-MSCs were either (re-) differentiated in micromass cell pellets or encapsulated as isolated cells in alginate hydrogels. Samples were treated with and without the gap junction inhibitor 18-α glycyrrhetinic acid (18αGCA). DNA and glycosaminoglycan (GAG) content and gene expression levels (collagen I/II/X, aggrecan, and connexin 43) were quantified at various time points. Protein localization was determined using immunofluorescence, and adenosine-5'-triphosphate (ATP) was measured in conditioned media. While GAG/DNA was higher in alginate compared with pellets for chondrocytes, there were no differences in chondrogenic gene expression between culture models. Gap junction blocking reduced collagen II and extracellular ATP in all chondrocyte cultures and in BM-MSC hydrogels. However, differentiation capacity was not abolished completely by 18αGCA. Connexin 43 levels were high throughout chondrocyte cultures and peaked only later during BM-MSC differentiation, consistent with the delayed response of BM-MSCs to 18αGCA. Alginate hydrogels and microtissues are equally suited culture platforms for the chondrogenic (re-)differentiation of expanded human articular chondrocytes and BM-MSCs. Therefore, reducing direct cell-cell contacts does not affect in vitro chondrogenesis. However, blocking gap junctions compromises cell differentiation, pointing to a prominent role for hemichannel function in this process. Therefore, scaffold design strategies that promote an increasing distance between single chondroprogenitor cells do not restrict their differentiation potential in tissue-engineered constructs.

Factors that influence Information Systems decisions and outcomes: A summary of key themes from four case studies" (2006). ACIS 2006 Proceedings. Paper 43.

This paper reports a summary of key findings from an examination of Information Systems decision making in four organisations. The study focused on what factors influenced decision makers during the critical preimplementation phase of Information Systems projects when systems were evaluated, selected and acquired. Using data gathered from interviews and organisational documentation, a critical hermeneutic analysis was performed in order to build an understanding of how informational and contextual influences acted on decision makers. Eight broad themes of factors were identified as having influence on decision makers and outcomes.

Clergy Sexual Abuse Litigation: Survivors Seeking Justice By Jennifer M. Balboni (Boulder: First Forum Press, 2011, 180pp. £43.77)

Balboni identifies her interest as being the processes of official disclosure and the path taken to civil litigation by survivors of child sexual abuse by Roman Catholic Clergy. The empirical data, on which this work is based, come in the form of in-depth face-to-face interviews with 22 survivors of clergy sexual abuse who have pursued litigation and 13 of their advocates. Balboni provides a space for survivors’ accounts of the ‘why’ behind their decision making and the impact of civil litigation on their lives to be heard, discussed and contextualized with both clarity and sensitivity. She acknowledges the breadth and depth of survivor responses, and the perspectives of their legal advocates, employing defiance theory, symbolic interaction and other points of analysis, to capture the journey of survivors towards litigation and beyond. Balboni’s work is deeply poignant in its recognition of survivors’ voices, the complex transformative capacity of litigation, the effects of community forming amongst survivors and the complex nature of ‘empowerment’ obtained by survivors through civil litigation. Acknowledging that, for many survivors, litigation becomes a means of identity change and truth telling, Balboni admits that ‘these survivors helped me understand that litigation is more about voice than monetary settlement’ (p. 149). This work is not deeply analytical or theoretically rich but privileges the voices of survivors and their advocates with sufficient frameworks to contextualize and explain participants’ perspectives and experiences.

The antigen 43 structure reveals a molecular velcro-like mechanism of autotransporter-mediated bacterial clumping

Aggregation and biofilm formation are critical mechanisms for bacterial resistance to host immune factors and antibiotics. Autotransporter (AT) proteins, which represent the largest group of outer-membrane and secreted proteins in Gram-negative bacteria, contribute significantly to these phenotypes. Despite their abundance and role in bacterial pathogenesis, most AT proteins have not been structurally characterized, and there is a paucity of detailed information with regard to their mode of action. Here we report the structure–function relationships of Antigen 43 (Ag43a), a prototypic self-associating AT protein from uropathogenic Escherichia coli. The functional domain of Ag43a displays a twisted L-shaped β-helical structure firmly stabilized by a 3D hydrogen-bonded scaffold. Notably, the distinctive Ag43a L shape facilitates self-association and cell aggregation. Combining all our data, we define a molecular “Velcro-like” mechanism of AT-mediated bacterial clumping, which can be tailored to fit different bacterial lifestyles such as the formation of biofilms.