Correct development of the vertebrate body plan requires the early definition of two asymmetric, perpendicular axes. The first axis is established during oocyte maturation, and the second is established by symmetry breaking shortly after fertilization. The physical processes generating the second asymmetric, or dorsal–ventral, axis are well understood, but the specific molecular determinants, presumed to be maternal gene products, are poorly characterized. Whilst enrichment of maternal mRNAs at the animal and vegetal poles in both the oocyte and the early embryo has been studied, little is known about the distribution of maternal mRNAs along either the dorsal–ventral or left–right axes during the early cleavage stages. Here we report an unbiased analysis of the distribution of maternal mRNA on all axes of the Xenopus tropicalis 8-cell stage embryo, based on sequencing of single blastomeres whose positions within the embryo are known. Analysis of pooled data from complete sets of blastomeres from four embryos has identified 908 mRNAs enriched in either the animal or vegetal blastomeres, of which 793 are not previously reported as enriched. In contrast, we find no evidence for asymmetric distribution along either the dorsal–ventral or left–right axes. We confirm that animal pole enrichment is on average distinctly lower than vegetal pole enrichment, and that considerable variation is found between reported enrichment levels in different studies. We use publicly available data to show that there is a significant association between genes with human disease annotation and enrichment at the animal pole. Mutations in the human ortholog of the most animally enriched novel gene, Slc35d1, are causative for Schneckenbecken dysplasia, and we show that a similar phenotype is produced by depletion of the orthologous protein in Xenopus embryos.

AimsGlucagon-like peptide-1 (GLP-1) receptor agonists and dipeptidyl peptidase-4 (DPP4) inhibitors improve glucose tolerance by still incompletely understood mechanisms. Each class of antihyperglycemic drugs has also been proposed to increase pancreatitis risk. Here, we compare systematically the effects of two widely-used GLP-1 analogues, liraglutide and exendin-4, and the DPP4 inhibitor, sitagliptin, in the mouse.MethodsC57BL6 mice were maintained for 131 days on a normal diet (ND) or a diet comprising 60% fat (HFD) before measurements of fasting blood glucose and insulin, and intraperitoneal glucose tolerance. Beta- and alpha- cell volume, and Reg3b immunoreactivity, were measured by immunohistochemical analysis of pancreatic slices.ResultsWhereas liraglutide (200 µg/kg) and exendin-4 (10 µg/kg) treatment reduced body weight and/or improved glucose tolerance, sitagliptin (10 mg/kg) was without effect on either parameter. Liraglutide caused a sharp reduction in beta-cell mass in both ND and HFD mice, whereas exendin-4 exerted no effect. By contrast, sitagliptin unmasked an action of high fat diet to increase beta-cell mass. Reg3B positive area was augmented by all three agents in normal chow-fed mice, whilst sitagliptin and exendin-4, but not liraglutide, affected this parameter in HFD animals. Correspondingly sitagliptin, but not the GLP-1 analogues, increased circulating amylase levels in ND and HFD mice.ConclusionsLiraglutide improves glucose tolerance in the mouse whilst exerting relatively modest effects on pancreatitis risk. Conversely, exendin-4 and sitagliptin, at doses which exert, respectively, minor or no effects on metabolic parameters, lead to signs of pancreatitis.

Objectives: Inherited RPE65-related retinal dystrophy is a rare, progressive disease, which until now has had no treatment options. Patients experience decreasing functional vision often from childhood and half of all patients are legally blind by age 18. This study evaluates the cost-effectiveness of voretigene neparvovec, a gene therapy which is the first treatment option ever developed, in a Danish setting. Methods: A Markov model with annual cycles was developed to describe the disease progression with and without treatment respectively. In the model, patients could go from moderate visual impairment through four additional states of visual impairment, with death as an option from all other states. Costs were based on list prices of drugs and either DRG costs or prices derived from a Danish eye care hospital clinic for cost of treatments. In line with Danish convention, the model focused on the perspective of a healthcare payer and thus, social costs were not included. Discount rates in Denmark follow national guidance in which the rate is 4% in the beginning with a lowering over time. The average rate, weighted by survivors in the model, ended up at 3,5% in the base case. In line with earlier NICE guidance, sensitivity analyses were performed also on lower discount rates given the upfront nature of payments. Results: In the base case, the ICER lands at DKK 834,241 with a QALY gain of 6.27. The results are mostly affected by choice of discount rate and assumptions regarding the duration of treatment effect. It is particularly worth noting that a lower discount rate of 1.5% almost halves the ICER, to DKK 447,034 with a QALY gain of 11.7. The undiscounted QALY gain is 20.1. Conclusions: With an assumed ICER threshold of DKK 1,000,000 for orphan indications, voretigene neparvovec is cost-effective in a Danish setting.

Intervention, Comparators, Outcomes, Settings, Time, Effects, Sensitivity) framework for an incremental cost effectiveness/utility, cost benefit, cost consequence, cost minimization, budget impact, willingness to pay, value of life or value of information analysis. FAE included sequential assessment of publications based on Pharmaceuticals Pricing Board, Consolidated Health Economic Evaluation Reporting Standards (CHEERS) and two earlier criteria. LoEE was built based on existing CCC classification (strong, moderate, minor/weak, no evidence) and extended significantly to cover HEEs. RESULTS: 41 publications fulfilled the search criteria and 15 publications were fully reviewed. In FAE, the most effort was used to assess the representativeness of patients, costs, quality of life and other inputs as well as modelling rationale for the Finnish setting. Generally, the applicability of foreign HEE results to the established Finnish CCC setting as such was poor and only one treatment had strong negative HEE evidence (multiple non-conflicting incremental HEE results). Most drugs had minor-to-moderate (one or conflicting) HEE evidence and three drugs had no valid HEE evidence. SLR, FAE and LoEE of HEEs was beneficial for the CCC. CONCLUSIONS: PICOSTEPS formed a working basis for the structural inclusion criteria to carry out the SLR. FAE was done based on the guidelines and demonstrated that there is significant need for local HEEs, especially if the published HEEs are rare or poorly generalizable to Finland. LoEE is an important part of review and should conclude the SLR statements based on HEEs.OBJECTIVES: Transfusion-dependent b-thalassemia (TDT) is a rare genetic disease that affects the production of healthy red blood cells. Conventional treatment involves regular blood transfusions and iron chelation, which is associated with potential adverse events and a substantial impact on quality of life. The only approved curative therapy is allogenic hematopoietic stem cell transplant (allo-HSCT), which carries the risk of complications, including graft-versus-host disease (GvHD). One novel treatment approach is gene therapy involving autologous stem cell transplant of the patient's own genetically modified hematopoietic stem cells. As new treatments for TDT are introduced, cost-utility analyses are needed to examine their value. The purpose of this study was to estimate utilities associated with treatment for TDT. METHODS: General population respondents in England valued eight health state vignettes (developed based on literature review and clinician, patient, and parent interviews) in time tradeoff interviews. There were two pre-transplant health states, three health states describing the year in which a transplant occurs, and three post-transplant health states. RESULTS: A total of 207 participants completed interviews (49.8% female; mean age ¼ 43.2 years; Newcastle, n¼87; London, n¼72; Bristol, n¼48). Mean (SD) utilities for the pre-transplant health states were 0.73 (0.25) for transfusion with oral chelation and 0.63 (0...

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