BACKGROUND: Despite considerable global investigation over several decades, the roles of vitamin D in health and disease development remains convoluted. One recognised issue is the difficulty of accurately measuring the active forms of vitamin D. Advances made include some new methods addressing the potential interference by excluding epimers and isobars. However, there is no evidence that epimers are without function. Therefore, the aim of this study was to develop and validate, for the first time, a new assay to simultaneously measure levels of 6 forms of vitamin D along with two epimers. The assay was applied to multilevel certified reference material calibrators and 25 pooled human sera samples obtained from the Vitamin D External Quality Assessment Scheme (DEQAS) to demonstrate its efficiency. RESULTS: The assay is capable of simultaneously measuring 8 vitamin D analogues over the calibration ranges and LODs (in nmol/L) of: 1alpha25(OH)2D2 [0.015-1; 0.01], 1alpha25(OH)2D3 [0.1-100; 0.01], 25OHD3 [0.5-100, 0.025], 3-epi-25OHD3 [0.1-100, 0.05], 25OHD2 [0.5-100, 0.025], 3-epi-25OHD2 [0.1-100, 0.05], vitamin D3 [0.5-100, 0.05] and vitamin D2 [0.5-100, 0.05], using stanozolol-d3 as internal standard. Certified reference material calibrators and external quality control samples (DEQAS) were analysed to meet the standards outlined by National Institute of Standards and Technology (NIST). Validation steps included recovery and both precision and accuracy under inter- and intra-day variation limit of detection, and analysis of each analyte over a linear range. All validation parameters were in line with acceptable Food and Drug Administration (FDA) guidelines and the standards of the National Institute of Standards and Technology (NIST). All eight analogues were quantified with the 25OHD levels being commensurate with DEQAS data. CONCLUSIONS: This report details the application of a new LC-MS/MS based assay for the efficient analysis of eight analogues of vitamin D over a range of samples, which is a significant advance over the existing methods. Simultaneous measure of 8 vitamin D analogues does not compromise the analytical capability of the assay to quantify the commonly used biomarker (25OHD) for vitamin D status. The results demonstrate the feasibility of applying the assay in research and clinical practice that i) excludes misleading measures owing to epimers and isobars and ii) is able to quantify the excluded component to facilitate further in vivo investigation into the roles of ubiquitous epimers.
Electromagnetic (EM) tracking systems are highly susceptible to field distortion. The interference can cause measurement errors up to a few centimeters in clinical environments, which limits the reliability of these systems. Unless corrected for, this measurement error imperils the success of clinical procedures. It is therefore fundamental to dynamically calibrate EM tracking systems and compensate for measurement error caused by field distorting objects commonly present in clinical environments. We propose to combine a motion model with observations of redundant EM sensors and compensate for field distortions in real-time. We employ a simultaneous localization and mapping (SLAM) technique to accurately estimate the pose of the tracked instrument while creating the field distortion map. We conducted experiments with 6 degrees-of-freedom motions in the presence of field distorting objects in research and clinical environments. We applied our approach to improve the EM tracking accuracy and compared our results to a conventional sensor fusion technique. Using our approach, the maximum tracking error was reduced by 67% for position measurements and by 64% for orientation measurements. Currently, clinical applications of EM trackers are hampered by the adverse distortion effects. Our approach introduces a novel method for dynamic field distortion compensation, independent from pre-operative calibrations or external tracking devices, and enables reliable EM navigation for potential applications.
People frequently fail to wait for delayed rewards after choosing them. These preference reversals are sometimes thought to reflect self-control failure. Other times, however, continuing to wait for a delayed reward may be counterproductive (e.g., when reward timing uncertainty is high). Research has demonstrated that people can calibrate how long to wait for rewards in a given environment. Thus, the role of self-control might be to integrate information about the environment to flexibly adapt behavior, not merely to promote waiting. Here we tested effects of acute stress, which has been shown to tax control processes, on persistence, and the calibration of persistence, in young adult human participants. Half the participants (n = 60) performed a task in which persistence was optimal, and the other half (n = 60) performed a task in which it was optimal to quit waiting for reward soon after each trial began. Each participant completed the task either after cold pressor stress or no stress. Stress did not influence persistence or optimal calibration of persistence. Nevertheless, an exploratory analysis revealed an “inverted-U” relationship between cortisol increase and performance in the stress groups, suggesting that choosing the adaptive waiting policy may be facilitated with some stress and impaired with severe stress.
In recent years, DNA origami nanorulers for superresolution (SR) fluorescence microscopy have been developed from fundamental proof-of-principle experiments to commercially available test structures. The self-assembled nanostructures allow placing a defined number of fluorescent dye molecules in defined geometries in the nanometer range. Besides the unprecedented control over matter on the nanoscale, robust DNA origami nanorulers are reproducibly obtained in high yields. The distances between their fluorescent marks can be easily analysed yielding intermark distance histograms from many identical structures. Thus, DNA origami nanorulers have become excellent reference and training structures for superresolution microscopy. In this work, we go one step further and develop a calibration process for the measured distances between the fluorescent marks on DNA origami nanorulers. The superresolution technique DNA-PAINT is used to achieve nanometrological traceability of nanoruler distances following the guide to the expression of uncertainty in measurement (GUM). We further show two examples how these nanorulers are used to evaluate the performance of TIRF microscopes that are capable of single-molecule localization microscopy (SMLM).
Portable Vis/NIRS are flexible tools for fast and unbiased analyses of constituents with minimal sample preparation. This study developed calibration models for dry matter content (DMC) and carotenoids in fresh cassava roots using a portable Vis/NIRS system. We examined the effects of eight data pre-treatment combinations on calibration models and assessed calibrations on processed and intact root samples. We compared Vis/NIRS derived-DMC to other phenotyping methods. The results of the study showed that the combination of standard normal variate and de-trend (SNVD) with first derivative calculated on two data points and no smoothing (SNVD+1111) was adequate for a robust model. Calibration performance was higher with processed than the intact root samples for all the traits although intact root models for some traits especially total carotenoid content (TCC) (R2c = 96%, R2cv = 90%, RPD = 3.6 and SECV = 0.63) were sufficient for screening purposes. Using three key quality traits as templates, we developed models with processed fresh root samples. Robust calibrations were established for DMC (R2c = 99%, R2cv = 95%, RPD = 4.5 and SECV = 0.9), TCC (R2c = 99%, R2cv = 91%, RPD = 3.5 and SECV = 2.1) and all Trans β-carotene (ATBC) (R2c = 98%, R2cv = 91%, RPD = 3.5 and SECV = 1.6). Coefficient of determination on independent validation set (R2p) for these traits were also satisfactory for ATBC (91%), TCC (88%) and DMC (80%). Compared to other methods, Vis/NIRS-derived DMC from both intact and processed roots had very high correlation (>0.95) with the ideal oven-drying than from specific gravity method (0.49). There was equally a high correlation (0.94) between the intact and processed Vis/NIRS DMC. Therefore, the portable Vis/NIRS could be employed for the rapid analyses of DMC and quantification of carotenoids in cassava for nutritional and breeding purposes.
Next-generation sequencing (NGS) provides a broad investigation of the genome, and it is being readily applied for the diagnosis of disease-associated genetic features. However, the interpretation of NGS data remains challenging owing to the size and complexity of the genome and the technical errors that are introduced during sample preparation, sequencing and analysis. These errors can be understood and mitigated through the use of reference standards - well-characterized genetic materials or synthetic spike-in controls that help to calibrate NGS measurements and to evaluate diagnostic performance. The informed use of reference standards, and associated statistical principles, ensures rigorous analysis of NGS data and is essential for its future clinical use.
The mixing of magmas is a common phenomenon in explosive eruptions. Concentration variance is a useful metric of this process and its decay (CVD) with time is an inevitable consequence during the progress of magma mixing. In order to calibrate this petrological/volcanological clock we have performed a time-series of high temperature experiments of magma mixing. The results of these experiments demonstrate that compositional variance decays exponentially with time. With this calibration the CVD rate (CVD-R) becomes a new geochronometer for the time lapse from initiation of mixing to eruption. The resultant novel technique is fully independent of the typically unknown advective history of mixing - a notorious uncertainty which plagues the application of many diffusional analyses of magmatic history. Using the calibrated CVD-R technique we have obtained mingling-to-eruption times for three explosive volcanic eruptions from Campi Flegrei (Italy) in the range of tens of minutes. These in turn imply ascent velocities of 5-8 meters per second. We anticipate the routine application of the CVD-R geochronometer to the eruptive products of active volcanoes in future in order to constrain typical “mixing to eruption” time lapses such that monitoring activities can be targeted at relevant timescales and signals during volcanic unrest.
People and societies seek to combat harmful events. However, because resources are limited, every wrong righted leaves another wrong left unchecked. Responses must therefore be calibrated to the magnitude of the harm. One underappreciated factor that affects this calibration may be people’s oversensitivity to intent. Across a series of studies, people saw intended harms as worse than unintended harms, even though the two harms were identical. This harm-magnification effect occurred for both subjective and monetary estimates of harm, and it remained when participants were given incentives to be accurate. The effect was fully mediated by blame motivation. People may therefore focus on intentional harms to the neglect of unintentional (but equally damaging) harms.
The certification of a new standard reference material for small-angle scattering [NIST Standard Reference Material (SRM) 3600: Absolute Intensity Calibration Standard for Small-Angle X-ray Scattering (SAXS)], based on glassy carbon, is presented. Creation of this SRM relies on the intrinsic primary calibration capabilities of the ultra-small-angle X-ray scattering technique. This article describes how the intensity calibration has been achieved and validated in the certified Q range, Q = 0.008-0.25 Å(-1), together with the purpose, use and availability of the SRM. The intensity calibration afforded by this robust and stable SRM should be applicable universally to all SAXS instruments that employ a transmission measurement geometry, working with a wide range of X-ray energies or wavelengths. The validation of the SRM SAXS intensity calibration using small-angle neutron scattering (SANS) is discussed, together with the prospects for including SANS in a future renewal certification.
Current biodiversity assessment and biomonitoring are largely based on the morphological identification of selected bioindicator taxa. Recently, several attempts have been made to use eDNA metabarcoding as an alternative tool. However, until now, most applied metabarcoding studies have been based on the taxonomic assignment of sequences that provides reference to morphospecies ecology. Usually, only a small portion of metabarcoding data can be used due to a limited reference database and a lack of phylogenetic resolution. Here, we investigate the possibility to overcome these limitations by using a taxonomy-free approach that allows the computing of a molecular index directly from eDNA data without any reference to morphotaxonomy. As a case study, we use the benthic diatoms index, commonly used for monitoring the biological quality of rivers and streams. We analysed 87 epilithic samples from Swiss rivers, the ecological status of which was established based on the microscopic identification of diatom species. We compared the diatom index derived from eDNA data obtained with or without taxonomic assignment. Our taxonomy-free approach yields promising results by providing a correct assessment for 77% of examined sites. The main advantage of this method is that almost 95% of OTUs could be used for index calculation, compared to 35% in the case of the taxonomic assignment approach. Its main limitations are under-sampling and the need to calibrate the index based on the microscopic assessment of diatoms communities. However, once calibrated, the taxonomy-free molecular index can be easily standardized and applied in routine biomonitoring, as a complementary tool allowing fast and cost-effective assessment of the biological quality of watercourses. This article is protected by copyright. All rights reserved.