Differing responses of osteogenic cell lines to β-glycerophosphate

Abstract Ascorbic acid (Asc), dexamethasone (Dex) and β-glycerophosphate (β-Gly) are commonly used to promote osteogenic behaviour by osteoblasts in vitro. According to the literature, several osteosarcoma cells lines appear to respond differently to the latter with regards to proliferation kinetics and osteogenic gene transcription. Unsurprisingly, these differences lead to contrasting data between publications that necessitate preliminary studies to confirm the phenotype of the chosen osteosarcoma cell line in the presence of Asc, Dex and β-Gly. The present study exposed Saos-2 cells to different combinations of Asc, Dex and β-Gly for 14 days and compared the response with immortalised human mesenchymal stromal/stem cells (MSCs). Cell numbers, cytotoxicity, mineralised matrix deposition and cell proliferation were analysed to assess osteoblast-like behaviour in the presence of Asc, Dex and β-Gly. Additionally, gene expression of runt-related transcription factor 2 (RUNX2); osteocalcin (OCN); alkaline phosphatase (ALP); phosphate regulating endopeptidase homolog X-linked (PHEX); marker of proliferation MKI67 and proliferating cell nuclear antigen (PCNA) was performed every two days during the 14-day cultures. It was found that proliferation of Saos-2 cells was significantly decreased by the presence of β-Gly which contrasted with hMSCs where no change was observed. Furthermore, unlike hMSCs, Saos-2 cells demonstrated an upregulated expression of late osteoblastic markers, OCN and PHEX that suggested β-Gly could affect later stages of osteogenic differentiation. In summary, it is important to consider that β-Gly significantly affects key cell processes of Saos-2 when using it as an osteoblast-like cell model

A duchenne muscular dystrophy gene hot spot mutation in dystrophin-deficient Cavalier King Charles Spaniels is amenable to exon 51 skipping

BACKGROUND Duchenne muscular dystrophy (DMD), which afflicts 1 in 3500 boys, is one of the most common genetic disorders of children. This fatal degenerative condition is caused by an absence or deficiency of dystrophin in striated muscle. Most affected patients have inherited or spontaneous deletions in the dystrophin gene that disrupt the reading frame resulting in unstable truncated products. For these patients, restoration of the reading frame via antisense oligonucleotide-mediated exon skipping is a promising therapeutic approach. The major DMD deletion "hot spot" is found between exons 45 and 53, and skipping exon 51 in particular is predicted to ameliorate the dystrophic phenotype in the greatest number of patients. Currently the mdx mouse is the most widely used animal model of DMD, although its mild phenotype limits its suitability in clinical trials. The Golden Retriever muscular dystrophy (GRMD) model has a severe phenotype, but due to its large size, is expensive to use. Both these models have mutations in regions of the dystrophin gene distant from the commonly mutated DMD "hot spot". METHODOLOGY/PRINCIPAL FINDINGS Here we describe the severe phenotype, histopathological findings, and molecular analysis of Cavalier King Charles Spaniels with dystrophin-deficient muscular dystrophy (CKCS-MD). The dogs harbour a missense mutation in the 5' donor splice site of exon 50 that results in deletion of exon 50 in mRNA transcripts and a predicted premature truncation of the translated protein. Antisense oligonucleotide-mediated skipping of exon 51 in cultured myoblasts from an affected dog restored the reading frame and protein expression. CONCLUSIONS/SIGNIFICANCE Given the small size of the breed, the amiable temperament and the nature of the mutation, we propose that CKCS-MD is a valuable new model for clinical trials of antisense oligonucleotide-induced exon skipping and other therapeutic approaches for DMD

Developmental Regulation of an Adhesin Gene during Cellular Morphogenesis in the Fungal Pathogen Candida albicans

Peer reviewedPublisher PD

Expanding medical education in general practice

General practitioners are major providers of medical education, with recent expansion at both undergraduate and postgraduate levels, particularly in rural areas. General practitioners not only teach general practice, but also provide access to patients well suited to other aspects of medical school curricula. Teaching demands on GPs, already high, may increase with the addition of extra medical school places, additional medical schools, and expanding vocational GP training. The challenge for this growth is the low morale in the GP workforce; a feeling of being under valued, a lack of trained GP teachers, and poor remuneration for teaching

The Brexit deal and UK fisheries—has reality matched the rhetoric?

Fisheries management has been a strongly contested aspect of the UK’s position in the EU since UK accession, with the fishing industry frequently questioning both the efficacy and fairness of arrangements. During the campaign for UK exit (Brexit) from the EU, and the subsequent negotiations of a new legal and political relationship from 2016 to 2020, senior UK political leaders strongly committed to deliver radically changed fisheries arrangements with respect to the three central issues: regulatory autonomy; access to waters; and quota shares, all while maintaining minimal trade impacts. The Trade and Cooperation Agreement diverges from this Brexit rhetoric. While some regulatory independence has been achieved, UK fisheries management continues in a state of interdependence and significant EU access to UK waters remains, even in the 6–12 nautical mile territorial waters. While the UK gained an increase in quota shares which is estimated to reach 107 thousand tonnes of landed weight annually by 2025 (an increase of 21.3% for quota species and 16.9% for all species, or 17.8% and 12.4% by value), this pales in comparison to the UK Government’s stated ambitions for zonal attachment (achieving 68% by weight and by value - a potential shortfall of 229,000 tonnes / £281 million). This modest change explains the negative reaction of the fishing industry and claims of betrayal in the face of the UK Government’s announcement of a “successful” deal. The stark delivery gap between rhetoric and reality means the UK government faces a challenging start to managing fisheries outside of the Common Fisheries Policy

A Risk Benefit Analysis of Mariculture as a means to Reduce the Impacts of Terrestrial Production of Food and Energy

The Scottish Aquaculture Research Forum (SARF) and WWF-UK commissioned this study to investigate whether the pressure on land and freshwater for future food and energy resources, and impacts on the climate, related to greenhouse gas (GHG) emissions, may be reduced through expansion of global mariculture. The study has undertaken a high level assessment of the ‘environmental footprint’ of global mariculture and terrestrial-based food and energy production systems through the collation and assessment of available Life Cycle Assessments (LCA) for key food products (beef, pork, chicken, freshwater finfish, marine finfish, shellfish and crustacean species) and biomass (terrestrial and algal) for energy production. The outputs of the footprint comparison were then used to assess the risks and benefits of increasing global mariculture, through the development of projected future scenarios in which mariculture contributes differing proportions of projected future food requirements. The analysis also qualitatively considered the socio-economic and wider environmental risks and benefits (e.g. in relation to ecosystem services) of global mariculture expansion, where expansion may occur geographically and whether future technological developments may help mitigate against identified impacts. The study identifies the key uncertainties and limitations of the risk/benefit analysis and makes prioritised recommendations on how these limitations can be addressed and the analysis developed for more regional or site-specific assessments

The growth of ‘Dry January’: promoting participation and the benefits of participation

This article explores contributors to the rapid growth of the annual UK alcohol abstinence challenge ‘Dry January’ and the benefits of registration. Evidence from four sources is presented: (i) registrations via the Dry January website, (ii) surveys of population-representative samples of drinkers, (iii) surveys of Dry January registrants and (iv) surveys of a control group of drinkers who wanted to change to their drinking behaviour but had not registered for Dry January. The data revealed that Dry January registrations increased 15-fold in 4 years. Participants reported that encouragement received from Dry January helped them to avoid drinking. Comparisons of Dry January registrants to the control group suggest that registering for Dry January reduced problematic drinking and enhanced the capacity to refuse alcohol. The four sources of data suggest that ‘social contagion’ and ‘diffusion’ have aided the growth of the awareness, appeal and practice of Dry January