Reliability and limitations of permanent tooth staging techniques.

Reliability, or repeatability, of permanent tooth staging techniques is usually expressed as Cohen's Kappa. This single value obscures information about the quantity and allocation of disagreements. In this study we assess and compare intra-observer reliability of permanent tooth staging techniques described by Nolla, Moorrees et al. and Demirjian et al. The sample was panoramic radiographs of healthy dental patients made up of 100 males and 100 females aged 6-15 years. All permanent teeth on the left side (excluding third molars) were scored twice. Weighted Kappa and percentage agreement were calculated. Results show Kappa values for all teeth combined as 0.918, 0.922 and 0.938 for Demirjian (number of teeth N = 2682), Nolla (N = 2698) and Moorrees (N = 2674) respectively. A comparison of Kappa values between upper and lower teeth showed marginally higher values for upper incisors and lower molar for all three scoring methods. Small differences in Kappa values were noted between tooth types with the upper first molar having smaller values than other teeth. Percentage agreement ranged from 81 % (Moorrees), 86 % (Nolla) to 87 % (Demirjian). Tooth stage differences between first and second assessments were not more than one stage. Our findings show that Demirjian scoring is marginally more reliable than Nolla or Moorrees. We suggest that (1) data for reliability are tabulated in full to show the quantity and allocation of disagreement between first and second readings, and (2) that the reliability sample is sufficiently large with a wide age range to include multiple different tooth stages.

Food system actor perspectives on future-proofing European food systems through plant breeding.

Crop improvement is a key innovation area in the pursuit of sustainable food systems. However, realising its potential requires integration of the needs and priorities of all agri-food chain stakeholders. In this study, we provide a multi-stakeholder perspective on the role of crop improvement in future-proofing the European food system. We engaged agri-business, farm- and consumer-level stakeholders, and plant scientists through an online survey and focus groups. Four of each group's top five priorities were shared and related to environmental sustainability goals (water, nitrogen and phosphorus efficiency, and heat stress). Consensus was identified on issues including considering existing alternatives to plant breeding (e.g. management strategies), minimising trade-offs, and addressing geographical variation in needs. We conducted a rapid evidence synthesis on the impacts of priority crop improvement options, highlighting the urgent need for further research examining downstream sustainability impacts to identify concrete targets for plant breeding innovation as a food systems solution.

Human iPSC-derived renal organoids engineered to report oxidative stress can predict drug-induced toxicity.

Advances in regenerative medicine have led to the construction of many types of organoids, which reproduce important aspects of endogenous organs but may be limited or disorganized in nature. While their usefulness for restoring function remains unclear, they have undoubted usefulness in research, diagnostics, and toxicology. In toxicology, there is an urgent need for better models for human kidneys. We used human iPS-cell (hiPSC)-derived renal organoids to identify HMOX1 as a useful marker of toxic stress via the oxidative stress pathway, and then constructed an HMOX1 reporter in hiPSCs. We used two forms of hiPSC-derived HMOX1-reporter renal organoids to probe their ability to detect nephrotoxicants in a panel of blind-coded compounds. Our results highlight the potential usefulness, and some limitations, of HMOX1-reporter renal organoids as screening tools. The results may guide development of similar stress-reporting organoid assays for other stem-cell-derived organs and tissues.

Modeling observations of solar coronal mass ejections with heliospheric imagers verified with the Heliophysics System Observatory.

We present an advance toward accurately predicting the arrivals of coronal mass ejections (CMEs) at the terrestrial planets, including Earth. For the first time, we are able to assess a CME prediction model using data over two thirds of a solar cycle of observations with the Heliophysics System Observatory. We validate modeling results of 1337 CMEs observed with the Solar Terrestrial Relations Observatory (STEREO) heliospheric imagers (HI) (science data) from 8 years of observations by five in situ observing spacecraft. We use the self-similar expansion model for CME fronts assuming 60° longitudinal width, constant speed, and constant propagation direction. With these assumptions we find that 23%-35% of all CMEs that were predicted to hit a certain spacecraft lead to clear in situ signatures, so that for one correct prediction, two to three false alarms would have been issued. In addition, we find that the prediction accuracy does not degrade with the HI longitudinal separation from Earth. Predicted arrival times are on average within 2.6 ± 16.6 h difference of the in situ arrival time, similar to analytical and numerical modeling, and a true skill statistic of 0.21. We also discuss various factors that may improve the accuracy of space weather forecasting using wide-angle heliospheric imager observations. These results form a first-order approximated baseline of the prediction accuracy that is possible with HI and other methods used for data by an operational space weather mission at the Sun-Earth L5 point.

Long-term increases in soil carbon due to ecosystem fertilization by atmospheric nitrogen deposition demonstrated by regional-scale modelling and observations.

Fertilization of nitrogen (N)-limited ecosystems by anthropogenic atmospheric nitrogen deposition (Ndep) may promote CO2 removal from the atmosphere, thereby buffering human effects on global radiative forcing. We used the biogeochemical ecosystem model N14CP, which considers interactions among C (carbon), N and P (phosphorus), driven by a new reconstruction of historical Ndep, to assess the responses of soil organic carbon (SOC) stocks in British semi-natural landscapes to anthropogenic change. We calculate that increased net primary production due to Ndep has enhanced detrital inputs of C to soils, causing an average increase of 1.2 kgCm-2 (c. 10%) in soil SOC over the period 1750-2010. The simulation results are consistent with observed changes in topsoil SOC concentration in the late 20th Century, derived from sample-resample measurements at nearly 2000 field sites. More than half (57%) of the additional topsoil SOC is predicted to have a short turnover time (c. 20 years), and will therefore be sensitive to future changes in Ndep. The results are the first to validate model predictions of Ndep effects against observations of SOC at a regional field scale. They demonstrate the importance of long-term macronutrient interactions and the transitory nature of soil responses in the terrestrial C cycle.

The utility of polarized heliospheric imaging for space weather monitoring.

A polarizing heliospheric imager is a critical next generation tool for space weather monitoring and prediction. Heliospheric imagers can track coronal mass ejections (CMEs) as they cross the solar system, using sunlight scattered by electrons in the CME. This tracking has been demonstrated to improve the forecasting of impact probability and arrival time for Earth-directed CMEs. Polarized imaging allows locating CMEs in three dimensions from a single vantage point. Recent advances in heliospheric imaging have demonstrated that a polarized imager is feasible with current component technology.Developing this technology to a high technology readiness level is critical for space weather relevant imaging from either a near-Earth or deep-space mission. In this primarily technical review, we developpreliminary hardware requirements for a space weather polarizing heliospheric imager system and outline possible ways to flight qualify and ultimately deploy the technology operationally on upcoming specific missions. We consider deployment as an instrument on NOAA's Deep Space Climate Observatory follow-on near the Sun-Earth L1 Lagrange point, as a stand-alone constellation of smallsats in low Earth orbit, or as an instrument located at the Sun-Earth L5 Lagrange point. The critical first step is the demonstration of the technology, in either a science or prototype operational mission context.

150years of macronutrient change in unfertilized UK ecosystems: Observations vs simulations.

Understanding changes in plant-soil C, N and P using data alone is difficult due to the linkages between carbon, nitrogen and phosphorus cycles (C, N and P), and multiple changing long-term drivers (e.g. climate, land-use, and atmospheric N deposition). Hence, dynamic models are a vital tool for disentangling these drivers, helping us understand the dominant processes and drivers and predict future change. However, it is essential that models are tested against data if their outputs are to be concluded upon with confidence. Here, a simulation of C, N and P cycles using the N14CP model was compared with time-series observations of C, N and P in soils and biomass from the Rothamsted Research long-term experiments spanning 150years, providing an unprecedented temporal integrated test of such a model. N14CP reproduced broad trends in soil organic matter (SOM) C, N and P, vegetation biomass and N and P leaching. Subsequently, the model was used to decouple the effects of land management and elevated nitrogen deposition in these experiments. Elevated N deposition over the last 150years is shown to have increased net primary productivity (NPP) 4.5-fold and total carbon sequestration 5-fold at the Geescroft Wilderness experiment, which was re-wilded to woodland in 1886. In contrast, the model predicts that for cropped grassland conditions at the Park Grass site, elevated N deposition has very little effect on SOM, as increases in NPP are diverted from the soil. More broadly, these results suggest that N deposition is likely to have had a large effect on SOM and NPP in northern temperate and boreal semi-natural grasslands and forests. However, in cropped and grazed systems in the same region, whilst NPP may have been supported in part by elevated N deposition, declines in SOM may not have been appreciably counteracted by increased N availability.

Calcium/NFAT signalling promotes early nephrogenesis.

A number of Wnt genes are expressed during, and are known to be essential for, early kidney development. It is typically assumed that their products will act through the canonical ß-catenin signalling pathway. We have found evidence that suggests canonical Wnt signalling is not active in the early nephrogenic metanephric mesenchyme, but instead provide expressional and functional evidence that implicates the non-canonical Calcium/NFAT Wnt signalling pathway in nephrogenesis. Members of the NFAT (Nuclear Factor Activated in T cells) transcription factor gene family are expressed throughout murine kidney morphogenesis and NFATc3 is localised to the developing nephrons. Treatment of kidney rudiments with Cyclosporin A (CSA), an inhibitor of Calcium/NFAT signalling, decreases nephron formation--a phenotype similar to that in Wnt4(-/-) embryos. Treatment of Wnt4(-/-) kidneys with Ionomycin, an activator of the pathway, partially rescues the phenotype. We propose that the non-canonical Calcium/NFAT Wnt signalling pathway plays an important role in early mammalian renal development and is required for complete MET during nephrogenesis, potentially acting downstream of Wnt4.

Feeding grape seed extract to horses: effects on health, intake and digestion.

A feeding trial involving four Thoroughbred race horses was undertaken to establish whether inclusion of grape seed extract (GSE) in the diet of horses undergoing mild exercise had any effects on their general health, intake and digestion. Supplementation with GSE had no effect on either feed or water intake of the horses and the supplement was readily palatable to the horses at all levels of inclusion. Feeding GSE caused no adverse effects in terms of animal health (temperature, pulse and respirations rates), and there were some positive effects related to a presumed alteration in fermentation in the hindgut. Feeding GSE increased faecal pH, changing from acid faeces (pH 6.6) when no GSE was fed to neutral faeces (pH 7.0) when 150 mg GSE/kg body weight (BW) was fed. In addition, blood glucose concentrations were significantly (P < 0.05) decreased when GSE was fed at 100 and 150 mg/kg BW (5.50 ± 0.26 and 5.32 ± 0.72 mmol/l, respectively) compared with the control diet (5.77 ± 0.31 mmol/l). The actual mechanisms causing these alterations are yet to be elucidated, but could have important implications for the prevention of acidosis.

von Willebrand factor: evidence for variable clearance in vivo according to Y/C1584 phenotype and ABO blood group.

BACKGROUND: The plasma von Willebrand factor (VWF) level (VWF:Ag) is known to correlate with the VWF Y/C1584 variation and with ABO blood group. The ratio of the VWF propeptide (VWFpp) to VWF:Ag and the ratio of coagulation factor VIII (FVIII:C) to VWF:Ag have previously been used as indicators of VWF clearance and/or secretion. OBJECTIVES AND METHODS: To investigate the mechanism underlying the relationship between VWF phenotype and VWF:Ag, the VWFpp/VWF:Ag ratio and FVIII:C/VWF:Ag ratio were determined for plasmas of phenotype Y/C1584, Y/Y1584, blood group O and blood group A (n = 50 for each set). The blood group O plasmas comprised two sets of 25 with low and high mean VWF levels (Low-O and High-O), respectively; similarly for group A (Low-A and High-A). RESULTS AND CONCLUSIONS: The VWFpp/VWF:Ag ratio was greater than 1 (unity) for Y/C1584 plasmas and significantly higher than for Y/Y1584 plasmas; however, the FVIII:C/VWF:Ag ratio was near unity for both and was not significantly different. These results are consistent with increased clearance for Y/C1584 VWF. Similarly, the VWFpp/VWF:Ag ratio and FVIII:C/VWF:Ag ratio in combination were consistent with increased VWF clearance in blood group O compared with blood group A, and in Low-O and Low-A, respectively, compared with High-O and High-A. The data indicate that in vivo C1584 and blood group O are associated with increased VWF clearance, and that clearance contributes to differing VWF level within a given blood group.

Spontaneous generation of self-organized solitary wave structures at earth's magnetopause.

Spontaneous formation of solitary wave structures has been observed in Earth's magnetopause, and is shown to be caused by the breakup of a zonal flow by the action of drift wave turbulence. Here we show matched observations and modeling of coherent, large-scale solitary electrostatic structures, generated during the interaction of short-scale drift wave turbulence and zonal flows at the Earth's magnetopause. The observations were made by the Cluster spacecraft and the numerical modeling was performed using the wave-kinetic approach to drift wave-zonal flow interactions. Good agreement between observations and simulations has been found, thus explaining the emergence of the observed solitary structures as well as confirming earlier theoretical predictions of their existence.

A home-based exercise programme improves ankle range of motion in long-term venous ulcer patients.

BACKGROUND AND OBJECTIVES: Decreased ankle mobility is associated with delayed healing of venous ulcers. The aim of this study was to assess the efficacy of a 24-week home-based exercise programme in increasing ankle range of motion and increasing the strength and endurance of the calf muscle. The secondary aim was to see whether subjects with venous ulcers were able to maintain a consistent exercise programme. METHODS: Eleven patients with long-standing venous ulcers were encouraged to undertake a thrice-weekly 5-10 min ankle exercise programme using elastic resistance bands and stretching in a 24-week single-arm pilot study. Ankle range of motion was measured monthly using a bi-plane ankle goniometer. Pain scores and participation levels were also recorded. RESULTS: There were significant improvements in ankle range of motion achieved at 12 and 24 weeks. Median pain scores (scale of 1-10) were reduced from 5.2 at baseline to 2.0 at weeks 12 and 24. CONCLUSION: These results show that a simple, home-based exercise programme is effective in achieving gains in ankle range of motion and may help to reduce pain. Good participation levels show that it is not only effective, but also feasible. A larger randomized controlled study is needed to show whether there is a positive effect on ulcer healing.

Innexins in the lobster stomatogastric nervous system: cloning, phylogenetic analysis, developmental changes and expression within adult identified dye and electrically coupled neurons.

Gap junctions play a key role in the operation of neuronal networks by enabling direct electrical and metabolic communication between neurons. Suitable models to investigate their role in network operation and plasticity are invertebrate motor networks, which are built of comparatively few identified neurons, and can be examined throughout development; an excellent example is the lobster stomatogastric nervous system. In invertebrates, gap junctions are formed by proteins that belong to the innexin family. Here, we report the first molecular characterization of two crustacean innexins: the lobster Homarus gammarus innexin 1 (Hg-inx1) and 2 (Hg-inx2). Phylogenetic analysis reveals that innexin gene duplication occurred within the arthropod clade before the separation of insect and crustacean lineages. Using in situ hybridization, we find that each innexin is expressed within the adult and developing lobster stomatogastric nervous system and undergoes a marked down-regulation throughout development within the stomatogastric ganglion (STG). The number of innexin expressing neurons is significantly higher in the embryo than in the adult. By combining in situ hybridization, dye and electrical coupling experiments on identified neurons, we demonstrate that adult neurons that express at least one innexin are dye and electrically coupled with at least one other STG neuron. Finally, two STG neurons display no detectable amount of either innexin mRNAs but may express weak electrical coupling with other STG neurons, suggesting the existence of other forms of innexins. Altogether, we provide evidence that innexins are expressed within small neuronal networks built of dye and electrically coupled neurons and may be developmentally regulated.

A method for cold storage and transport of viable embryonic kidney rudiments.

Organ culture of mouse embryonic kidneys is a powerful system for studying normal renal development and for researching the developmental effects of experimental perturbations (drugs, antibodies, interfering RNA, and so on). In standard protocols, embryonic kidneys are isolated by delicate micro-dissection and placed in organ culture as soon as possible after the death of the donor mouse, before there is time to genotype them or to transport them elsewhere. This report demonstrates that fully viable embryonic kidneys can be isolated and cultured from crudely dissected caudal portions of embryos that have been stored on ice or at 4 degrees C for several days. This very simple technique can save considerable resources in laboratories that culture kidneys of transgenic mice: (i) cold storage allows embryos to be genotyped before their kidneys are cultured, and (ii) cold transport allows kidney research laboratories to study kidneys of transgenic mice raised elsewhere without the need for expensive importing of the mouse line itself. It will therefore substantially augment the ability of kidney research labs to perform pilot experiments on large numbers of different transgenic animals and to collaborate in new ways.

Designing follow-up intervals.

When an examination is needed to determine if an event has occurred there will be a loss of efficiency in using the resulting interval-censored data instead of the exact event times. In designing follow-up intervals this loss for longer intervals needs to be weighed against extra visits required by shorter intervals. We obtain results to quantify this for the estimation of the median and mean survival and for covariates in parametric regression models with equally spaced examination times. Asymptotic information loss for the log-normal and Weibull distributions are similar when comparisons are made between corresponding members of the two families. For distributions with coefficients of variation of 50 per cent or more, a choice of interval from 0.25 to 0.7 times the median is recommended.

Real-time scanning tunnelling microscopy imaging of protein motion at electrode surfaces.

A mutant (K27C) of the blue copper protein azurin [Eur. J. Biochem. 194 (1990) 109; J. Mol. Biol. 221 (1991) 765] for orientated immobilisation on gold surfaces was analysed by scanning tunnelling microscopy (STM) both in a resting state and following the application of a short potential pulse between the tip and sample.