Progesterone analogue protects stressed photoreceptors via bFGF-mediated calcium influx.

Retinitis pigmentosa (RP) is a degenerative retinal disease leading to photoreceptor cell loss. In 2011, our group identified the synthetic progesterone ‘Norgestrel’ as a potential treatment for RP. Subsequent research showed Norgestrel to work through progesterone receptor membrane component 1 (PGRMC1) activation and upregulation of neuroprotective basic fibroblast growth factor (bFGF). Using trophic factor deprivation of 661W photoreceptor-like cells, we aimed to further elucidate the mechanism leading to Norgestrel-induced neuroprotection. In the present manuscript, we show by flow cytometry and live-cell immunofluorescence that Norgestrel induces an increase in cytosolic calcium in both healthy and stressed 661Ws over 24h. Specific PGRMC1 inhibition by AG205 (1 μM) showed this rise to be PGRMC1-dependent, primarily utilising calcium from extracellular sources, for blockade of L-type calcium channels by verapamil (50 μM) prevented a Norgestrel-induced calcium influx in stressed cells. Calcium influx was also shown to be bFGF-dependent, for siRNA knock down of bFGF prevented Norgestrel-PGRMC1 induced changes in cytosolic calcium. Notably, we demonstrate PGRMC1-activation is necessary for Norgestrel-induced bFGF upregulation. We propose that Norgestrel protects through the following pathway: binding to and activating PGRMC1 expressed on the surface of photoreceptor cells, PGRMC1 activation drives bFGF upregulation and subsequent calcium influx. Importantly, raised intracellular calcium is critical to Norgestrel's protective efficacy, for extracellular calcium chelation by EGTA abrogates the protective effects of Norgestrel on stressed 661W cells in vitro.

Mapping and Displaying Structural Transformations between XML and PDF

Documents are often marked up in XML-based tagsets to delineate major structural components such as headings, paragraphs, figure captions and so on, without much regard to their eventual displayed appearance. And yet these same abstract documents, after many transformations and 'typesetting' processes, often emerge in the popular format of Adobe PDF, either for dissemination or archiving. Until recently PDF has been a totally display-based document representation, relying on the underlying PostScript semantics of PDF. Early versions of PDF had no mechanism for retaining any form of abstract document structure but recent releases have now introduced an internal structure tree to create the so called 'Tagged PDF'. This paper describes the development of a plugin for Adobe Acrobat which creates a two-window display. In one window is shown an XML document original and in the other its Tagged PDF counterpart is seen, with an internal structure tree that, in some sense, matches the one seen in XML. If a component is highlighted in either window then the corresponding structured item, with any attendant text, is also highlighted in the other window. Important applications of correctly Tagged PDF include making PDF documents reflow intelligently on small screen devices and enabling them to be read out in correct reading order, via speech synthesiser software, for the visually impaired. By tracing structure transformation from source document to destination one can implement the repair of damaged PDF structure or the adaptation of an existing structure tree to an incrementally updated document.