PEGylation of lysine residues reduces the pro-migratory activity of IGF-I

Background: The insulin-like growth factor (IGF) system is composed of ligands and receptors which regulate cell proliferation, survival, differentiation and migration. Some functions are regulated via intracellular signaling cascades, others by involvement of the extracellular milieu, including binding proteins and other extracellular matrix proteins. However, understanding of their functions and the exact nature of these interactions remains incomplete. Methods: IGF-I was PEGylated at its lysine sites - K27, K65 and K68. Binding of PEG-IGF-I to the IGFBPs was analyzed using BIAcore and its ability to activate the IGF-IR was assessed using IGF-IR phosphorylation assay. Furthermore, functional consequences of PEGylating the lysine residues of IGF-I was investigated using cell viability and cell migration assays. In addition, particular downstream signaling pathways regularly implicated in these mechanisms were also dissected using phospho-AKT and phospho-ERK1/2 assays. Results: In this study, IGF-I specifically PEGylated at lysine 27 (PEG-K27), 65 (PEG-K65) or 68 (PEG-K68) were employed. Receptor phosphorylation was only reduced by 2-fold with PEG-K65 and PEG-K68 over all the time points tested, and as observed in two cell types, 3T3 fibroblasts and MCF-7 breast cancer cells. PEGylation at K27 resulted in a much larger effect, with more than 10-fold lower activation for 3T3 fibroblasts and a ~3 fold reduced IGF-IR activation for MCF-7 breast cancer cells over 15 minutes. In addition, all PEG-IGF-I variants demonstrated a ten-fold reduction in the association rate to IGF binding proteins (IGFBPs). Functionally, all PEG variants completely lost their ability to induce cell migration in the presence of IGFBP-3/vitronectin (VN) complexes as compared to IGF-I; in contrast, cell viability was fully preserved. Further investigations into the downstream signaling pathways revealed that the PI3-K/AKT pathway was preferentially affected upon treatment with the PEG-IGF-I variants compared to the MAPK/ERK pathway. Conclusion: PEGylation of IGF-I has an impact on cell migration but not cell viability. General significance: PEG-IGF-I may differentially modulate IGF-I mediated functions that are dependent on its interaction with its receptor as well as key extracellular proteins such as VN and IGFBPs.

Intimate perceptions : exploring justice from the perspective of Australian survivors of Clergy Sexual Violence

The issue of child sexual abuse in Christian institutions has been persistent and politicalised across the world. Images and stories of abusive clergy, and their superiors who protect them, are common fodder for commercial and public media. In November 2012 the Australian Prime Minister announced a Royal Commission into child sexual abuse in Australian institutions. This came on the back of multiple calls such an inquiry. At this same time in Victoria, Australia, a Parliamentary Inquiry in the same issue was completing its process and preparing a report. This study draws on submissions made to the Victorian Parliamentary Inquiry and data from 15 ethnographic interviews with survivors of child sexual abuse in Christian institutions of Australia. The common themes of these sources are of betrayal, grief, a persistent search for justice and for recognition of the trauma rendered, not only to the lives of survivors but also to their families and communities. These are not new themes in the literature of child sexual abuse in Christian Institutions, however the perceptions of victimisation in the Australian context has only been explored in limited ways.

What is a “Christian” jurisprudence?

The term ‘jurisprudence’ is derived etymologically from the Latin juris, meaning law, and prudentia, meaning wisdom. So jurisprudence simply means the wisdom of the law, or, as it has come to mean in scholarly legal circles these days, the theory of law. It asks fundamental questions regarding the nature and definition of law. And so, the question I wish to pose to us today is what does a truly “Christian” theory of law look like? One that is faithful to, as taken from the conference brochure, the “historic Christian faith” in its “principles and practice”. To contextualise this question, I must give you a deceptively brief and superficial overview of prevailing theories of law – and for those of you who know more about the topic, I apologise for the crass nature of my summary – time prevents me from doing any more...

Is Dawkins a modern-day Nicodemus?

This article applies a Wittgensteinian approach to the examination of the intelligibility of religious belief, in the wake of the recent attack on the Judeo-Christian religion by Richard Dawkins's book The God Delusion. The article attempts to show that Dawkins has confused religion with superstition, and that while Dawkins's arguments are decisive in the case of superstition, they do not successfully show religion to be a delusion. Religious belief in God is not like belief in the existence of a planet, and genuine religious faith is not like the belief in something for which there is not yet enough evidence, like belief in dark matter. The Christian doctrines of the resurrection and eternal life are misconstrued if they are understood as factual claims because they are then merely shallow superstitions, and not the great religious riddles they are meant to be.