The use of time-resolved fluorescence imaging in the study of protein kinase C localisation in cells

Background: Two-photon-excitation fluorescence lifetime imaging (2P-FLIM) was used to investigate the association of protein kinase C alpha (PKCα) with caveolin in CHO cells. PKCα is found widely in the cytoplasm and nucleus in most cells. Upon activation, as a result of increased intracellular Ca2+ and production of DAG, through G-protein coupled-phospholipase C signalling, PKC translocates to a variety of regions in the cell where it phosphorylates and interacts with many signalling pathways. Due to its wide distribution, discerning a particular interaction from others within the cell is extremely difficult. Results: Fluorescence energy transfer (FRET), between GFP-PKCα and DsRed-caveolin, was used to investigate the interaction between caveolin and PKC, an aspect of signalling that is poorly understood. Using 2P-FLIM measurements, the lifetime of GFP was found to decrease (quench) in certain regions of the cell from ~2.2 ns to ~1.5 ns when the GFP and DsRed were sufficiently close for FRET to occur. This only occurred when intracellular Ca2+ increased or in the presence of phorbol ester, and was an indication of PKC and caveolin co-localisation under these conditions. In the case of phorbol ester stimulated PKC translocation, as commonly used to model PKC activation, three PKC areas could be delineated. These included PKCα that was not associated with caveolin in the nucleus and cytoplasm, PKCα associated with caveolin in the cytoplasm/perinuclear regions and probably in endosomes, and PKC in the peripheral regions of the cell, possibly indirectly interacting with caveolin. Conclusion: Based on the extent of lifetime quenching observed, the results are consistent with a direct interaction between PKCα and caveolin in the endosomes, and possibly an indirect interaction in the peripheral regions of the cell. The results show that 2P-FLIM-FRET imaging offers an approach that can provide information not only confirming the occurrence of specific protein-protein interactions but where they occur within the cell

Book Review: Natural Resource Management and Sustainable Development: How Ghana’s Petroleum Can Create Sustainable Economic Prosperity

Kwamina Panford. Palgrave, Macmillan, 2017, 249 pp. ISBN:  978-1-137-54071-3 (hard back) Reviewed by Karl Botchwa

Arbitrating natural resource disputes: Current and future trends

نظراً للهبوط الحاد في أسعار السلع الأساسية حول العالم، وبالأخص الوقود الحفري، يحاول المستثمرون وغيرهم من الأطراف المعنية إعادة ترتيب استثماراتهم ومصالحهم وإعادة تنظيمها وإعادة تموضعها. والسؤال الذي يطرح نفسه هو: مَن الذي يتحمل تكاليف أو أعباء الخسائر التي يتكبدونها في استثماراتهم وفي إعادة تموضعها؟ من المتوقع أن يؤدي هذا التوزيع أو إعادة التوازن للتكاليف والخسائر إلى نشوء نزاعات تفضي بدورها إلى التحكيم. فمن المرجح أن تكون عمليات الاندماج المقترحة التي قد تفشل، والبيوت والآبار التي قد يتم تركها، وشركاء المشاريع المشتركة الذين قد لا يُوفون بالتزاماتهم، والتغطية التأمينية التي قد لا يتم تنفيذها، وما إلى ذلك، أن تكون مثاراً للنزاعات. وهذه النزاعات ستكون إلى جانب نزاعات الموارد الطبيعية التي تتمحور حول منازعات التأميم، ونقل الحقوق، والتمويل، والتنمية، والتشغيل، والتنظيم والفساد، والمنازعات البيئية والاجتماعية، التي تحدث في مجال الموارد الطبيعية. وسوف تستعرض هذه الورقة النزاعات القائمة وكذلك النزاعات الناشئة أو المتوقعة في مجال استثمار الموارد. وهي تَخلُص إلى إن التحكيم لا يزال أحد أكثر الطرق فعالية في تسوية هذه النزاعات. ومع ذلك، فسوف تكون هناك العديد من التحديات. وهذه التحديات سوف تطعن في شرعية التحكيم باعتباره الوسيلة الأنسب لتسوية النزاعات. ويُعزى ذلك إلى كون قضايا الإفلاس والملكية الفكرية أو ملكية البيانات، والانتهاكات التنظيمية العامة والإهمال لا تصلح بسهولة للتحكيم. وفوق ذلك، فإن بعض الدفوع من قبيل القوة القاهرة، وتعذر التنفيذ، والضرورة، والحصانة السيادية التي تُثار في مثل هذه المنازعات قد يثبت أنها تمثل إشكالية أمام التحكيم.Owing to the precipitous slump in world commodity prices, particularly fossil fuel, investors and other stakeholders have been trying to re-arrange, re-organize and re-position their investments and interests. The question that arises is: who bears the costs or burden of losses in the investments and the re-positioning? This allocation or rebalancing of costs and losses is predicted to generate disputes which would result in arbitration. Proposed mergers that may fall through, blocks and wells that may be abandoned, joint-venture partners that may not meet their obligations, insurance cover that may not be honored, etc., are likely to be sources of disputes. These will be in addition to “normal” resource disputes that revolve around resource nationalism, rights transfers, financing, development, operating, regulatory, corruption, environmental and social disputes, which occur in the natural resource business. This paper will explore the existing as well as the emerging or expected disputes in resource investment. It will argue that arbitration remains one of the most effective ways of resolving these disputes. However, there will be challenges. These challenges will challenge the very legitimacy of arbitration as the most appropriate forum or means of resolving the disputes. This is because issues of bankruptcy, intellectual property or data ownership, public regulatory violations and negligence do not lend themselves easily to arbitration. Furthermore, some of the defenses such as force majeure, impossibility of performance, necessity and sovereign immunity to be raised in such disputes may prove problematic for arbitration

Sharing individual patient and parasite-level data through the WorldWide Antimalarial Resistance Network platform: A qualitative case study.

BACKGROUND: Increasingly, biomedical researchers are encouraged or required by research funders and journals to share their data, but there's very little guidance on how to do that equitably and usefully, especially in resource-constrained settings. We performed an in-depth case study of one data sharing pioneer: the WorldWide Antimalarial Resistance Network (WWARN). METHODS: The case study included a records review, a quantitative analysis of WAARN-related publications, in-depth interviews with 47 people familiar with WWARN, and a witness seminar involving a sub-set of 11 interviewees. RESULTS: WWARN originally aimed to collate clinical, in vitro, pharmacological and molecular data into linked, open-access databases intended to serve as a public resource to guide antimalarial drug treatment policies. Our study describes how WWARN navigated challenging institutional and academic incentive structures, alongside funders' reluctance to invest in capacity building in malaria-endemic countries, which impeded data sharing. The network increased data contributions by focusing on providing free, online tools to improve the quality and efficiency of data collection, and by inviting collaborative authorship on papers addressing policy-relevant questions that could only be answered through pooled analyses. By July 1, 2016, the database included standardised data from 103 molecular studies and 186 clinical trials, representing 135,000 individual patients. Developing the database took longer and cost more than anticipated, and efforts to increase equity for data contributors are on-going. However, analyses of the pooled data have generated new methods and influenced malaria treatment recommendations globally. Despite not achieving the initial goal of real-time surveillance, WWARN has developed strong data governance and curation tools, which are now being adapted relatively quickly for other diseases. CONCLUSIONS: To be useful, data sharing requires investment in long-term infrastructure. To be feasible, it requires new incentive structures that favour the generation of reusable knowledge