The use of fluorescence microscopy to define polymer localisation to the late endocytic compartments in cells that are targets for drug delivery

Macromolecular therapeutics and nano-sized drug delivery systems often require localisation to specific intracellular compartments. In particular, efficient endosomal escape, retrograde trafficking, or late endocytic/lysosomal activation are often prerequisites for pharmacological activity. The aim of this study was to define a fluorescence microscopy technique able to confirm the localisation of water-soluble polymeric carriers to late endocytic intracellular compartments. Three polymeric carriers of different molecular weight and character were studied: dextrin (Mw~50,000 g/mol), a N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer (Mw approximately 35,000 g/mol) and polyethylene glycol (PEG) (Mw 5000 g/mol). They were labelled with Oregon Green (OG) (0.3-3 wt.%; <3% free OG in respect of total). A panel of relevant target cells were used: THP-1, ARPE-19, and MCF-7 cells, and primary bovine chondrocytes (currently being used to evaluate novel polymer therapeutics) as well as NRK and Vero cells as reference controls. Specific intracellular compartments were marked using either endocytosed physiological standards, Marine Blue (MB) or Texas-red (TxR)-Wheat germ agglutinin (WGA), TxR-Bovine Serum Albumin (BSA), TxR-dextran, ricin holotoxin, C6-7-nitro-2,1,3-benzoxadiazol-4-yl (NBD)-labelled ceramide and TxR-shiga toxin B chain, or post-fixation immuno-staining for early endosomal antigen 1 (EEA1), lysosomal-associated membrane proteins (LAMP-1, Lgp-120 or CD63) or the Golgi marker GM130. Co-localisation with polymer-OG conjugates confirmed transfer to discreet, late endocytic (including lysosomal) compartments in all cells types. The technique described here is a particularly powerful tool as it circumvents fixation artefacts ensuring the retention of water-soluble polymers within the vesicles they occupy.

Economic crisis and public services: note 1, December 2008

The paper examines the impact of the economic crisis on public services, including government reponses and implications for companies operating in public services.

FARM-Africa South Africa Northern Cape land reform and advocacy programme 2004-2008: final evaluation

FARM-Africa South Africa has played a crucial and important role in filling the gap that existed after the return of land to communities by government in the Northern Cape. Their support to farmer communities during the post-settlement phase has been critical for making productive use of land. During 2004-2008, FARM-SA has worked in 20 community projects in the Northern Cape, benefiting 745 poor households.

Memorandum submitted by the University of Greenwich: are the consumer price increases for electricity imposed in January 2008 justified?

This paper examines whether increases to published wholesale prices justify the retail electricity price increases imposed on residential consumers in January 2008. The study is based on analysis of two questions: Is the reported wholesale price a reliable indicator of the cost electricity retailers are paying to buy power; and is the corporate structure of the British electricity sector competitive? [Taken from first paragraph of summary]

Spank performance tour [at National Review of Live Art 2008]

Spank is a duet about two women’s journeys as they play out different roles. One woman recalls stinging anecdotes marked by intimacy, jealousy, fear and fantasy from her childhood in the 70s suburbs – round the corner from where Mike Leigh would film Life Is Sweet. Her Double (projected on screen) reads from a newspaper about an event that punctured the local community, creating an enduring cultural scar, her voice lacking emotion. Both women must eventually face one another and ask, who is deceiving whom? The consequences of guilt and shame and the unstable shift from private memory to public consequence is inevitable and deadly.[ABSTRACT BY THE AUTHOR]