The chick embryo; a new drug delivery model for neuroblastoma

Neuroblastoma (NB) commonly presents as high risk disease which despite intensive multimodal therapies is often fatal. Preclinical models are required to aid the development of novel therapeutics for this challenging childhood malignancy, however current systems are complex and inherently costly. We aimed to explore the ability of the chick embryo to act as a new tool for NB therapeutic research. "High risk"- MYCN amplified human neuroblastoma cells were xenografted on to the surface of the chick embryo chorioallantoic membrane (CAM) and tumours allowed to form over a 7 day period. qPCR was used to detect the effects of a differentiation agent, retinoic acid (RA), firstly on NB cells in culture, and then on NB tumours in the chick embryo model. Tumours formed in the chick embryo model 4 days after the introduction of NB cells on to the CAM. After 7 days, analysis of fully formed tumours demonstrated active proliferation and vascular recruitment from the surrounding CAM. In culture, RA induced morphological changes consistent with the differentiation of NB cells. RT-qPCR identified reproducible changes in gene expression in response to RA. Increased expression of differentiation markers ROBO2 and STMN4 and decreased expression of the stem cell marker KLF4 was observed in cell culture. Similar changes in the expression of these genes were also seen during in vivo chick embryo experiments. Against expectations, levels of the MYCN transcription factor did not fall significantly following 3 days of RA treatment during in vitro or in vivo experimentation. These results suggest that other targets may also be involved in the RA induced differentiation of these cells. We have observed the ability of the chick embryo to act as an in vivo model system for NB therapeutic research. Reproducible changes in gene expression induced by the administration of retinoic acid have been detected using this model

Photograph of gravestone of Ellen Grace Mather, West Hobart, Tasmania

Photograph of gravestone of Ellen Grace Mather taken in the Quaker (Society of Friends) Burial Ground established in 1836 at Mellifont Street, West Hobart, Tasmania until it was closed in 1912. The gravestones have been moved to the edge of what is now the Friends' Park, a recreational park and children's playground. Inscription reads: Ellen Grace Mather Daughter of TB and EG Mather - Died 16th of the 9th Month 1895. Aged 14 Days'

Flamingo Vol. II N 4

Chaparral. Taking It Seriously . Prose. 1. Whiz Bang. Our History Lesson . Prose. 1. Ubersax, Delmar. Untitled. Picture. 4. Mather, William G. Frozen Fields . Poem. 5. A.F.T. Open Season . Prose. 6. Anonymous. Bob . Poem. 6. Anonymous. Untitled. Poem. 6. Mather, William G. Untitled. Poem. 6. R. Song . Poem. 7. Q. Woman\u27s Love . Poem. 7. Holt, Kilburn. A Lost Art . Poem. 7. C.B. A Sonnet . Poem. 7. Keeler, Clyde. Untitled. Picture. 9. Keeler, Clyde. Mid-Years . Picture. 0. Anonymous. Inside Stuff . Prose. 9. Anonymous. Untitled. prose. 9. Keeler, Clyde. untitled. Picture. 10. Anonymous. Untitled. Prose. 10. Anonymous. The First One: A Flapper\u27s Lament . Prose. 11. Keeler, Clyde. Untitled. Picture. 11. Keeler, Clyde. Untitled. Prose. 11. Schmitz, Edward. Untitled. Picture. 11. W.A.V. The Microbes . Picture. 11. Williams, Grace. Heaven\u27s Bells! Picture. 12. Anonymous. Heaven\u27s Bells! Prose. 12. Anonymous. Tragedy Picture. 12. Anonymous. Untitled. Picture. 12. Anonymous. Famous Hells Picture. 12. Anonymous. Denison\u27s Hall of Fame: Uncle Sam Brierly . Prose. 13. Anonymous. Untitled. Prose. 14. Anonymous. Anxious Moments . Cartoon. 16. Keeler, Clyde. Untitled. Picture. 18. Anonymous. Untitled. Prose. 18. Williams, Grace. Just An ordinary C-Man . Picture. 18. Anonymous. As The Denisonian Reporter Would Have Done it . Prose. 19. Anonymous. Untitled. Prose. 19. Anonymous. Not The Morning After-But The Same Night . Prose. 20. Ubersax, Delmar. Untitled. Picture. 20. Anonymous. Untitled. Prose. 20. Anonymous. Untitled. Picture. 20. Anonymous. Not A Flirtation . Prose. 20. Anonymous. Crossing The Bar . Poem. 20. Anonymous. Untitled. Prose. 22. Williams, Grace. Untitled. Picture. 23. Anonymous. Untitled. Prose. 23. Williams, Grace. Untitled. Picture. 24. Anonymous. Untitled. Prose. 24. Anonymous. Benny Says . Prose. 25. Jester. Thy Name is Woman . Prose. 25. W.G.K. An Ode to A Rabbit . Poem. 26. Anonymous. Untitled. Poem. 26. Anonymous. Untitled. Poem. 27. Keeler, Clyde. Untitled. Picture. 27. Y. Catastrophe . Poem. 28. Ed. Untitled. Picture. 29. Beanpot. We Know Her, Too . Prose. 29. Puppet. Anonymous. Prose. 30. Benapot. Untitled. Prose. 30. Wasp. Untitled. Prose. 30. Whiz Bang. Untitled. Prose. 30. Chaparral. A Flapper . Poem. 31. Whiz Bang. Untitled. Prose. 31. Jester. Untitled. Prose. 31. Reel, Virginia. Untitled. Prose. 32. O\u27Lantern, Jack. Untitled. Prose. 32. Sun Dodger. Untitled. Prose. 32. Beanpot. Untitled. Prose. 32. Jester. Untitled. Prose. 32

Optimising the chick chorioallantoic membrane xenograft model of neuroblastoma for drug delivery

Background Neuroblastoma is a paediatric cancer that despite multimodal therapy still has a poor outcome for many patients with high risk tumours. Retinoic acid (RA) promotes differentiation of some neuroblastoma tumours and cell lines, and is successfully used clinically, supporting the view that differentiation therapy is a promising strategy for treatment of neuroblastoma. To improve treatment of a wider range of tumour types, development and testing of novel differentiation agents is essential. New pre-clinical models are therefore required to test therapies in a rapid cost effective way in order to identify the most useful agents. Methods As a proof of principle, differentiation upon ATRA treatment of two MYCN-amplified neuroblastoma cell lines, IMR32 and BE2C, was measured both in cell cultures and in tumours formed on the chick chorioallantoic membrane (CAM). Differentiation was assessed by 1) change in cell morphology, 2) reduction in cell proliferation using Ki67 staining and 3) changes in differentiation markers (STMN4 and ROBO2) and stem cell marker (KLF4). Results were compared to MLN8237, a classical Aurora Kinase A inhibitor. For the in vivo experiments, cells were implanted on the CAM at embryonic day 7 (E7), ATRA treatment was between E11 and E13 and tumours were analysed at E14. Results Treatment of IMR32 and BE2C cells in vitro with 10 μM ATRA resulted in a change in cell morphology, a 65% decrease in cell proliferation, upregulation of STMN4 and ROBO2 and downregulation of KLF4. ATRA proved more effective than MLN8237 in these assays. In vivo, 100 μM ATRA repetitive treatment at E11, E12 and E13 promoted a change in expression of differentiation markers and reduced proliferation by 43% (p < 0.05). 40 μM ATRA treatment at E11 and E13 reduced proliferation by 37% (p < 0.05) and also changed cell morphology within the tumour. Conclusion Differentiation of neuroblastoma tumours formed on the chick CAM can be analysed by changes in cell morphology, proliferation and gene expression. The well-described effects of ATRA on neuroblastoma differentiation were recapitulated within 3 days in the chick embryo model, which therefore offers a rapid, cost effective model compliant with the 3Rs to select promising drugs for further preclinical analysis

Flamingo Vol. II N 2

Keeler, Clyde. Dad\u27s Day . Picture. 0. Puppet. Untitled. Prose. 1. Crescent. Untitled. Prose. 1. Williams, Charles L. As To Reading Biography . Prose. 5. Verlaine, Paul. Kyrie Eleison . Poem. 6. Anonymous. Mystery . Poem. 6. R. To You . Poem. 7. Q. The Passion Flower . Poem. 7. Bennett, G.W. November . Poem. 7. R. Storm . Poem. 7. Anonymous. Untitled.Prose. 8. Anonymous. No Doubt! . Poem. 8. Anonymous. Untitled. Picture. 8. McCutcheon, Dorothy. Ready-Made Reputations . Prose. 9. Anonymous. Untitled. Prose. 9. Anonymous. Untitled. Prose. 10. Ubersax, Delmar. Untitled. Prose. 10. Anonymous. Statistics From The Football Centers . Prose. 10. Anonymous. Our Own Handbook . Prose. 11. Anonymous. Real Reels . Prose. 12. Anonymous. Tire Trouble at The Round Table . Prose. 12. Anonymous. Untitled. Prose. 14. Anonymous. The Return From Dayton . Cartoon. 16. Anonymous. Untitled. Prose. 18. Anonymous. Down The Street . Poem. 18. Anonymous. The 7:30 . Poem. 18. Lord Jeff. In The Hole . Poem. 18. Anonymous. How It\u27s Done . Prose. 19. Anonymous. A Zoological Dream . Picture. 19. Anonymous. A Smooth Line . Prose. 20. Anonymous. Untitled. Prose. 20. Anonymous. You\u27ve Heart it . Prose. 20. Anonymous. Dog Gone! . Prose. 21. Anonymous. Untitled. Prose. 21. Cols Dis. Untitled. Poem. 21. Anonymous. Eddie Tor\u27s Lament . Poem. 22. Anonymous. Untitled. Prose. 22. Anonymous. The Long And Short of it. Prose. 22. Anonymous. Final Exam. In Zetetic Jactation 613 (You Look it up.) . Prose. 24. Pope of Fools. Love Letters of a Sceptic . Prose. 25. Anonymous. Untitled. Prose. 26. Williams, Grace. Untitled. Picture. 28. Anonymous. Untitled. Prose. 28. Goblin. It is a Wise Landlady . Prose. 28. Anonymous. Untitled. Prose. 28. Drexard. Untitled. Prose. 29. Chaparral. Untitled. Prose. 29. Punch Bowl. The Trifler . Prose. 29. Anonymous. His Finger Got Stuck in the Bowling Ball . Picture. 30. Chaparral. Untitled. Prose. 30. Banter. Untitled. Prose. 30. Tiger. Untitled. Prose. 30. Brown Jug. Untitled. Prose. 31. Octopus. Untitled. Prose. 31. Burr. Untitled. Prose. 31. Squib. Untitled. Prose. 31. Ubersax, Delmar. Untitled. Picture. 32. Purple Cow. Untitled. Prose. 32. Mather, William G. A Ditty . Poem. 12

Perspectives on enhancing physical activity and diet for health promotion among at-risk urban UK South Asian communities: a qualitative study

Objectives To explore perspectives on enhancing physical activity and diet among South Asians in urban deprived communities at high risk of chronic disease and to inform development of culturally appropriate health promotion intervention. Design Qualitative study using semistructured one-to-one and family group interviews with thematic analysis of data. Setting Urban disadvantaged communities in the East Midlands of the UK. Participants 45 respondents, including 34 people of South Asian origin (16 at-risk individuals, six family groups involving 18 relatives), of mainly Pakistani and Indian origin, including 16 non-English speakers; and 11 health professionals working locally with communities of concern. Results South Asian participants underlined the challenges of requiring family members across generations to engage in modifying dietary behaviours, and the central role of communal eating of traditional ‘Asian’ food in their cultural lives. Barriers to increasing physical activity included cost, personal safety and lack of time outside of long working hours and carer commitments. However, increasing walking activity was regarded as feasible by both community and health professional participants. Respondents emphasised using a social approach for potential interventions, undertaking activity with family or friends and with bilingual community peers to facilitate engagement, motivation and support. Spoken content and delivery of interventions was favoured, including personal stories and multilingual audio–visual information; within local informal rather than provider settings, including the home; and aided by pedometers for self-monitoring. Conclusions Focusing on physical activity by increasing walking may hold promise as health promotion in this deprived South Asian community context. Further intervention development, with exploration of feasibility and acceptability of the social approach and elements suggested, is merited

Contrasting long-term dynamics of antimicrobial resistance and virulence plasmids in Salmonella Typhimurium from animals

Plasmids are mobile elements that can carry genes encoding traits of clinical concern, including antimicrobial resistance (AMR) and virulence. Population-level studies of Enterobacterales, including Escherichia coli, Shigella and Klebsiella, indicate that plasmids are important drivers of lineage expansions and dissemination of AMR genes. Salmonella Typhimurium is the second most common cause of salmonellosis in humans and livestock in the UK and Europe. The long-term dynamics of plasmids between S. Typhimurium were investigated using isolates collected through national surveillance of animals in England and Wales over a 25-year period. The population structure of S. Typhimurium and its virulence plasmid (where present) were inferred through phylogenetic analyses using whole-genome sequence data for 496 isolates. Antimicrobial resistance genes and plasmid markers were detected in silico. Phenotypic plasmid characterization, using the Kado and Liu method, was used to confirm the number and size of plasmids. The differences in AMR and plasmids between clades were striking, with livestock clades more likely to carry one or more AMR plasmid and be multi-drug-resistant compared to clades associated with wildlife and companion animals. Multiple small non-AMR plasmids were distributed across clades. However, all hybrid AMR-virulence plasmids and most AMR plasmids were highly clade-associated and persisted over decades, with minimal evidence of horizontal transfer between clades. This contrasts with the role of plasmids in the short-term dissemination of AMR between diverse strains in other Enterobacterales in high-antimicrobial-use settings, with implications for predicting plasmid dissemination amongst S. Typhimurium

Spatial reconstruction of Scottish summer temperatures from tree rings

The authors wish to thank The Carnegie Trust for the Universities of Scotland for providing funding for Miloš Rydval’s PhD. The Scottish pine network expansion has been an ongoing task since 2006 and funding must be acknowledged to the following projects: EU project ‘Millennium’ (017008-2), Leverhulme Trust project ‘RELiC: Reconstructing 8000 years of Environmental and Landscape change in the Cairngorms (F/00 268/BG)’ and the NERC project ‘SCOT2K: Reconstructing 2000 years of Scottish climate from tree rings (NE/K003097/1)’. We also thank Rider University for a faculty research fellowship that supported Daniel Druckenbrod, and Paul J. Krusic for compiling and making available a PC version of the point-by-point PCR software. Lamont-Doherty Earth Observatory contribution No. 8032.A detailed understanding of past temporal patterns and spatial expression of temperature variations is important to place recent anthropogenic climate change into a longer term context. In order to fill the current gap in our understanding of northwest European temperature variability, point-by-point principal component regression was used to reconstruct a spatial field of 0.5° temperature grids across Scotland. A sequence of reconstructions utilizing several combinations of detrending and disturbance correction procedures, and a selection of tree-ring parameters [including ring width (RW), maximum latewood density (MXD) and blue intensity (BI)] was used in an evaluation of reconstruction skill. The high resolution of the reconstructed field serves also as a diagnostic tool to spatially assess the temperature reconstruction potential of local chronologies. Best reconstruction results, reaching calibration r2 = 65.8% and verification r2 = 63.7% in central Scotland over the 1901–1976 period, were achieved using disturbance-corrected and signal-free detrended RW chronologies merged with BI data after low-pass (high-pass) filtering the RW (BI) chronologies. Calibration and verification r2 > 50% was attained for central, north and east Scotland, >40% in west and northwest, and >30% in southern Scotland with verification of nearly all grids showing some reconstruction skill. However, the full calibration potential of reconstructions outside central Scotland was reduced either due to residual disturbance trends undetected by the disturbance correction procedure or due to other climatic or non-climatic factors which may have adversely affected the strength of the climate signal.PostprintPeer reviewe

Evaluating the Effects of SARS-CoV-2 Spike Mutation D614G on Transmissibility and Pathogenicity.

Global dispersal and increasing frequency of the SARS-CoV-2 spike protein variant D614G are suggestive of a selective advantage but may also be due to a random founder effect. We investigate the hypothesis for positive selection of spike D614G in the United Kingdom using more than 25,000 whole genome SARS-CoV-2 sequences. Despite the availability of a large dataset, well represented by both spike 614 variants, not all approaches showed a conclusive signal of positive selection. Population genetic analysis indicates that 614G increases in frequency relative to 614D in a manner consistent with a selective advantage. We do not find any indication that patients infected with the spike 614G variant have higher COVID-19 mortality or clinical severity, but 614G is associated with higher viral load and younger age of patients. Significant differences in growth and size of 614G phylogenetic clusters indicate a need for continued study of this variant

The genetic architecture of the human cerebral cortex

The cerebral cortex underlies our complex cognitive capabilities, yet little is known about the specific genetic loci that influence human cortical structure. To identify genetic variants that affect cortical structure, we conducted a genome-wide association meta-analysis of brain magnetic resonance imaging data from 51,665 individuals. We analyzed the surface area and average thickness of the whole cortex and 34 regions with known functional specializations. We identified 199 significant loci and found significant enrichment for loci influencing total surface area within regulatory elements that are active during prenatal cortical development, supporting the radial unit hypothesis. Loci that affect regional surface area cluster near genes in Wnt signaling pathways, which influence progenitor expansion and areal identity. Variation in cortical structure is genetically correlated with cognitive function, Parkinson's disease, insomnia, depression, neuroticism, and attention deficit hyperactivity disorder