Investigation of the role of long non-coding RNAs in oncogene induced cellular senescence

Cellular senescence is a stable arrest of cell proliferation induced by several factors such as activated oncogenes, oxidative stress and shortening of telomeres. Senescence acts as a tumour suppression mechanism to halt the progression of cancer. However, senescence may also impact negatively upon tissue regeneration, thus contributing to the effects of ageing. The eukaryotic genome is controlled by various modes of transcriptional and translational regulation. Focus has therefore centred on the role of long non- coding RNAs (lncRNAs) in regulating the genome. Accordingly, understanding how lncRNAs function to regulate the senescent genome is integral to improving our knowledge and understanding of tumour suppression and ageing. Within this study, I set out to investigate the expression of lncRNAs’ expression within models of senescence. Through a custom expression array, I have shown that expression of multiple different lncRNAs is up-regulated and down regulated in IMR90 replicative senescent fibroblasts and oncogene-induced senescent melanocytes. LncRNA expression was determined to be specific to stable senescence-associated cell arrest and predominantly within the nucleus of senescent cells. In order to examine the function of lncRNA expression in senescence, I selected lncRNA transcript ENST0000430998 (lncRNA_98) to focus my investigations upon. LncRNA_98 was robustly upregulated within multiple models of senescence and efficiently depleted using anti-sense oligonucleotide technology. Characterisation and unbiased RNA-sequencing of lncRNA_98 deficient senescent cells highlighted a list of genes that are regulated by lncRNA_98 expression in senescent cells and may regulate aspects of the senescence program. Specifically, the formation of SAHF was impeded upon depletion of lncRNA_98 expression and levels of total pRB protein expression severely decreased. Validation and recapitulation of consequences of pRB depletion was confirmed through lncRNA_98 knock-out cells generated using CRISPR technology. Surprisingly, inhibition of ATM kinase functions permitted the restoration of pRB protein levels within lncRNA_98 deficient cells. I propose that lncRNA_98 antagonizes the ability of ATM kinase to downregulate pRB expression at a post-transcriptional level, thereby potentiating senescence. Furthermore, lncRNA expression was detected within fibroblasts of old individuals and visualised within senescent melanocytes in human benign nevi, a barrier to melanoma progression. Conversely, mining of 337 TCGA primary melanoma data sets highlighted that the lncRNA_98 gene and its expression was lost from a significant proportion of melanoma samples, consistent with lncRNA_98 having a tumour suppressor functions. The data presented in this study illustrates that lncRNA_98 expression has a regulatory role over pRB expression in senescence and may regulate aspects of tumourigenesis and ageing.

Spiritual guides: supporting adults who nurture the spirituality of children

This research considers the role of those who support voluntary youth leaders in their responsibility to nurture spiritual development in children and young people. It discusses particularly those who work in organisations with an ethos of supporting such spiritual development and at the same time of welcoming members of many faiths and encouraging their full participation in their various faith traditions. The specific context of the research is the work of the Guide Movement and, in particular, of Girlguiding in the United Kingdom. The research addresses the current challenges but also the enormous value of providing for young people a multi-faith space in which it is genuinely ‘OK to do God’. It discusses the issues which leaders are raising and some suggestions which trainers have made for addressing these.

Perceptions of climate change, environmental variability and the role of agricultural adaptation strategies by small-scale farmers in Africa: the case of Mwanga district in Northern Tanzania

Abstract The potential impacts of climate change and environmental variability are already evident in most parts of the world, which is witnessing increasing temperature rates and prolonged flood or drought conditions that affect agriculture activities and nature-dependent livelihoods. This study was conducted in Mwanga District in the Kilimanjaro region of Tanzania to assess the nature and impacts of climate change and environmental variability on agriculture-dependent livelihoods and the adaptation strategies adopted by small-scale rural farmers. To attain its objective, the study employed a mixed methods approach in which both qualitative and quantitative techniques were used. The study shows that farmers are highly aware of their local environment and are conscious of the ways environmental changes affect their livelihoods. Farmers perceived that changes in climatic variables such as rainfall and temperature had occurred in their area over the period of three decades, and associated these changes with climate change and environmental variability. Farmers’ perceptions were confirmed by the evidence from rainfall and temperature data obtained from local and national weather stations, which showed that temperature and rainfall in the study area had become more variable over the past three decades. Farmers’ knowledge and perceptions of climate change vary depending on the location, age and gender of the respondents. The findings show that the farmers have limited understanding of the causes of climatic conditions and environmental variability, as some respondents associated climate change and environmental variability with social, cultural and religious factors. This study suggests that, despite the changing climatic conditions and environmental variability, farmers have developed and implemented a number of agriculture adaptation strategies that enable them to reduce their vulnerability to the changing conditions. The findings show that agriculture adaptation strategies employ both planned and autonomous adaptation strategies. However, the study shows that increasing drought conditions, rainfall variability, declining soil fertility and use of cheap farming technology are among the challenges that limit effective implementation of agriculture adaptation strategies. This study recommends further research on the varieties of drought-resilient crops, the development of small-scale irrigation schemes to reduce dependence on rain-fed agriculture, and the improvement of crop production in a given plot of land. In respect of the development of adaptation strategies, the study recommends the involvement of the local farmers and consideration of their knowledge and experience in the farming activities as well as the conditions of their local environment. Thus, the findings of this study may be helpful at various levels of decision making with regard to the development of climate change and environmental variability policies and strategies towards reducing farmers’ vulnerability to current and expected future changes.