Investigating the Vascularization of Tissue-Engineered Bone Constructs Using Dental Pulp Cells and 45S5 Bioglass(®) Scaffolds.

Identification of a suitable cell source combined with an appropriate 3D scaffold is an essential prerequisite for successful engineering of skeletal tissues. Both osteogenesis and angiogenesis are key processes for bone regeneration. This study investigated the vascularization potential of a novel combination of human dental pulp stromal cells (HDPSCs) with 45S5 Bioglass(®) scaffolds for tissue-engineered mineral constructs in vivo and in vitro. 45S5 Bioglass scaffolds were produced by the foam replication technique with the standard composition of 45 wt% SiO2, 24.5 wt% Na2O, 24.5 wt% CaO, and 6 wt% P2O5. HDPSCs were cultured in monolayers and on porous 45S5 Bioglass scaffolds under angiogenic and osteogenic conditions for 2-4 weeks. HDPSCs expressed endothelial gene markers (CD34, CD31/PECAM1, and VEGFR2) under both conditions in the monolayer. A combination of HDPSCs with 45S5 Bioglass enhanced the expression of these gene markers. Positive immunostaining for CD31/PECAM1 and VEGFR2 and negative staining for CD34 supported the gene expression data, while histology revealed evidence of endothelial cell-like morphology within the constructs. More organized tubular structures, resembling microvessels, were seen in the constructs after 8 weeks of implantation in vivo. In conclusion, this study suggests that the combination of HDPSCs with 45S5 Bioglass scaffolds offers a promising strategy for regenerating vascularized bone grafts

Full-depth temperature trends in the Northeastern Atlantic through the early 21st century

The vertical structure of temperature trends in the northeastern Atlantic (NEA) is investigated from a blend of Argo and hydrography data. The representativeness of sparse hydrography sampling in the basin-mean is assessed using a numerical model. Between 2003 and 2013, the NEA underwent a strong surface cooling (0-450?m) and a significant warming at intermediate and deep levels (1000?m-3000?m) that followed a strong cooling trend observed between 1988 and 2003. During 2003-2013, gyre-specific changes are found in the upper 1000?m (warming and cooling of the subtropical and subpolar gyres, respectively) whilst the intermediate and deep warming primarily occurred in the subpolar gyre, with important contributions from isopycnal heave and water mass property changes. The full-depth temperature change requires a local downward heat flux of 0.53?±?0.06?W?m?2 through the sea-surface, and its vertical distribution highlights the likely important role of the NEA in the recent global warming hiatus

The fallacy of placing confidence in confidence intervals

Interval estimates – estimates of parameters that include an allowance for sampling uncertainty – have long been touted as a key component of statistical analyses. There are several kinds of interval estimates, but the most popular are confidence intervals (CIs): intervals that contain the true parameter value in some known proportion of repeated samples, on average. The width of confidence intervals is thought to index the precision of an estimate; CIs are thought to be a guide to which parameter values are plausible or reasonable; and the confidence coefficient of the interval (e.g., 95 %) is thought to index the plausibility that the true parameter is included in the interval. We show in a number of examples that CIs do not necessarily have any of these properties, and can lead to unjustified or arbitrary inferences. For this reason, we caution against relying upon confidence interval theory to justify interval estimates, and suggest that other theories of interval estimation should be used instead

Labrador Sea subsurface density as a precursor of multidecadal variability in the North Atlantic: a multi-model study

The subpolar North Atlantic (SPNA) is a region with prominent decadal variability that has experienced remarkable warming and cooling trends in the last few decades. These observed trends have been preceded by slow-paced increases and decreases in the Labrador Sea density (LSD), which are thought to be a precursor of large-scale ocean circulation changes. This article analyses the interrelationships between the LSD and the wider North Atlantic across an ensemble of coupled climate model simulations. In particular, it analyses the link between subsurface density and the deep boundary density, the Atlantic Meridional Overturning Circulation (AMOC), the subpolar gyre (SPG) circulation, and the upper-ocean temperature in the eastern SPNA. All simulations exhibit considerable multidecadal variability in the LSD and the ocean circulation indices, which are found to be interrelated. LSD is strongly linked to the strength of the subpolar AMOC and gyre circulation, and it is also linked to the subtropical AMOC, although the strength of this relationship is model-dependent and affected by the inclusion of the Ekman component. The connectivity of LSD with the subtropics is found to be sensitive to different model features, including the mean density stratification in the Labrador Sea, the strength and depth of the AMOC, and the depth at which the LSD propagates southward along the western boundary. Several of these quantities can also be computed from observations, and comparison with these observation-based quantities suggests that models representing a weaker link to the subtropical AMOC might be more realistic

Test-retest reproducibility of a food frequency questionnaire (FFQ) and estimated effects on disease risk in the Norwegian Women and Cancer Study (NOWAC)

BACKGROUND: The Norwegian Women and Cancer Study (NOWAC) is a national population-based cohort study with 102 443 women enrolled at age 30–70 y from 1991 to 1997. The present study was a methodological sub-study to assess the test-retest reproducibility of the NOWAC food frequency questionnaire (FFQ), and to study how measurement errors in the data can affect estimates of disease risk. METHODS: A random sample of 2000 women aged 46–75 y was drawn from the cohort in 2002. A self-instructive health and lifestyle questionnaire with a FFQ section was mailed to the same subjects twice (test-retest), about three months apart, with a response rate of 75%. The FFQ was designed to assess habitual diet over the past year. We assess the reproducibility of single questions, food groups, energy, and nutrients with several statistical measures. We also demonstrate the method of regression calibration to correct disease risk estimates for measurement error. Alcohol intake (g/day) and high blood pressure (yes/no) is used in the example. RESULTS: For single foods there were some indications of seasonal reporting bias. For food groups and nutrients the reliability coefficients ranged from 0.5–0.8, and Pearson's r, Spearman's r(s), and two intraclass correlation coefficients gave similar results. Although alcohol intake had relatively high reproducibility (r = 0.72), odds ratio estimates for the association with blood pressure were attenuated towards the null value compared to estimates corrected by regression calibration. CONCLUSION: The level of reproducibility observed for the FFQ used in the NOWAC study is within the range reported for similar instruments, but may attenuate estimates of disease risk

Spin‐up of UK Earth System Model 1 (UKESM1) for CMIP6

For simulations intended to study the influence of anthropogenic forcing on climate, temporal stability of the Earth's natural heat, freshwater and biogeochemical budgets is critical. Achieving such coupled model equilibration is scientifically and computationally challenging. We describe the protocol used to spin‐up the UK Earth system model (UKESM1) with respect to pre‐industrial forcing for use in the 6th Coupled Model Intercomparison Project (CMIP6). Due to the high computational cost of UKESM1's atmospheric model, especially when running with interactive full chemistry and aerosols, spin‐up primarily used parallel configurations using only ocean/land components. For the ocean, the resulting spin‐up permitted the carbon and heat contents of the ocean's full volume to approach equilibrium over ~5000 years. On land, a spin‐up of ~1000 years brought UKESM1's dynamic vegetation and soil carbon reservoirs towards near‐equilibrium. The end‐states of these parallel ocean‐ and land‐only phases then initialised a multi‐centennial period of spin‐up with the full Earth system model, prior to this simulation continuing as the UKESM1 CMIP6 pre‐industrial control (piControl). The realism of the fully‐coupled spin‐up was assessed for a range of ocean and land properties, as was the degree of equilibration for key variables. Lessons drawn include the importance of consistent interface physics across ocean‐ and land‐only models and the coupled (parent) model, the extreme simulation duration required to approach equilibration targets, and the occurrence of significant regional land carbon drifts despite global‐scale equilibration. Overall, the UKESM1 spin‐up underscores the expense involved and argues in favour of future development of more efficient spin‐up techniques

Dying for change: A roadmap to refine the fish acute toxicity test after 40 years of applying a lethal endpoint

The fish acute toxicity test (TG203; OECD, 2019) is frequently used and highly embedded in hazard and risk assessment globally. The test estimates the concentration of a chemical that kills 50% of the fish (LC50) over a 96 h exposure and is considered one of the most severe scientific procedures undertaken. Over the years, discussions at the Organisation for Economic Co-operation and Development (OECD) have resulted in changes to the test which reduce the number of fish used, as well as the development of a (potential) replacement test (TG236, OECD, 2013). However, refinement of the mortality endpoint with an earlier (moribundity) endpoint was not considered feasible during the Test Guideline’s (TG) last update in 2019. Several stakeholders met at a UK-based workshop to discuss how TG203 can be refined, and identified two key opportunities to reduce fish suffering: (1) application of clinical signs that predict mortality and (2) shortening the test duration. However, several aspects need to be addressed before these refinements can be adopted. TG203 has required recording of major categories of sublethal clinical signs since its conception, with the option to record more detailed signs introduced in the 2019 update. However, in the absence of guidance, differences in identification, recording and reporting of clinical signs between technicians and laboratories is likely to have generated piecemeal data of varying quality. Harmonisation of reporting templates, and training in clinical sign recognition and recording are needed to standardise clinical sign data. This is critical to enable robust data-driven detection of clinical signs that predict mortality. Discussions suggested that the 96 h duration of TG203 cannot stand up to scientific scrutiny. Feedback and data from UK contract research organisations (CROs) conducting the test were that a substantial proportion of mortalities occur in the first 24 h. Refinement of TG203 by shortening the test duration would reduce suffering (and test failure rate) but requires a mechanism to correct new results to previous 96 h LC50 data. The actions needed to implement both refinement opportunities are summarised here within a roadmap. A shift in regulatory assessment, where the 96 h LC50 is a familiar base for decisions, will also be critical

In vitro and in vivo delivery of a sustained release nanocarrier-based formulation of an MRTF/SRF inhibitor in conjunctival fibrosis

BACKGROUND: Sustained drug delivery is a large unmet clinical need in glaucoma. Here, we incorporated a Myocardin-Related Transcription Factor/Serum Response Factor inhibitor, CCG-222740, into slow release large unilamellar vesicles derived from the liposomes DOTMA (1,2-di-O-octadecenyl-3-trimethylammonium propane) and DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine), and tested their effects in vitro and in vivo. RESULTS: The vesicles were spherical particles of around 130 nm and were strongly cationic. A large amount of inhibitor could be incorporated into the vesicles. We showed that the nanocarrier CCG-222740 formulation gradually released the inhibitor over 14 days using high performance liquid chromatography. Nanocarrier CCG-222740 significantly decreased ACTA2 gene expression and was not cytotoxic in human conjunctival fibroblasts. In vivo, nanocarrier CCG-222740 doubled the bleb survival from 11.0 ± 0.6 days to 22.0 ± 1.3 days (p = 0.001), decreased conjunctival scarring and did not have any local or systemic adverse effects in a rabbit model of glaucoma filtration surgery. CONCLUSIONS: Our study demonstrates proof-of-concept that a nanocarrier-based formulation efficiently achieves a sustained release of a Myocardin-Related Transcription Factor/Serum Response Factor inhibitor and prevents conjunctival fibrosis in an established rabbit model of glaucoma filtration surgery