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.

Malay Kingship in Kedah : Religion, Trade, and Society

The book probes and examines traditional sources of royal power and control, as well as indigenous socio-political systems in the Malay world. It is focused on the north-western Malaysian Sultanate of Kedah which is acknowledged as the oldest unbroken independent kingship line in the ‘Malay and Islamic world’ with 1,000 years of history. Little scholarly attention has been paid to its pre-modern history, society, religion, system of government and unique geographic situation, potentially controlling both land and sea lines of communication into the remainder of Southeast Asia. It will thus provide the first comprehensive treatment in English, or other languages, on Kedah’s pre-modern and nineteenth century historiography and can provide a foundation for comparative studies of the various Malay states which is presently lacking. The proposed book also sheds much needed light on a range of important topics in Malay history including: Kedah and the northern Melaka Straits history, colonial expansion and rivalry, Southeast Asian history and politics, interregional migration and the influence of the sea peoples or orang laut, traditional Malay socio-political and economic life, Islamic influences and the course of Thai-Malay relations. The book attempts to offer a new understanding, not only of Kedah, but of the political and cultural development of the entire Malay world and of its relationships with the broader forces in both its continental and maritime settings. It argues that Kedah does not seem to follow, and in fact, often seems to contradict what has been commonly been accepted as the “typical model” of the traditional Malay state. Thus it concludes that the ruling dynasty has historically exploited a wide range of unique environmental conditions, local traditions, global spiritual trends and economic forces to preserve and strengthen its political position. The scope and theme of book The Kedah Sultanate is the oldest unbroken independent kingship lines in the “Malay world” with 1,000 years of history, and arguably one of the oldest in the Islamic world. In this study I examine key geopolitical and spiritual attributes of Malay kingship that have traditionally cemented the ruler, the peoples, and the environment. Brief description of the primary audience for the book: There is little written in English or Malay on Kedah’s pre twentieth century history. The available sources only look at certain aspects of Kedah’s history, are outdated or are confined to a specific period often outside the scope of the book. It is therefore anticipated that the readership and market for the book includes: • Scholars of Southeast Asian history, Islam, kingship, trade. • Academics & Historians (including: Asian, Thai history, Islamic, Maritime, Persian, South Asian, Southeast Asian and Colonial) • Libraries • Students, particularly those in Malaysia (especially the states of Kedah, Perlis and Penang), Thailand and Singapore. • Universities • Scholars and students in Political Science & International Relations

Origins of Australian Football: Victoria's Early History. Volume 2 : A golden era begins : Football in 'Marvellous Melbourne', 1877 to 1885

This is the second volume of a five volume series that describes, assesses, and analyses football in Victoria during the nineteenth century. This volume looks at the cultural contexts of the sport in the late 1870s and early 1880s, describes the important matches played, and provides a full statistical account of this time period. This book is the first comprehensive discussion of the early period in Australian football's development.