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Untargeted metabolite profiling of biological samples is a challenge for analytical science due to the high degree of complexity of biofluids. Isobaric species may also not be resolved using mass spectrometry alone. As a result of these factors, many potential biomarkers may not be detected or are masked by co-eluting interferences in conventional LC-MS metabolomic analyses. In this study, a comprehensive liquid chromatography-mass spectrometry workflow incorporating a fast-scanning miniaturised high-field asymmetric waveform ion mobility spectrometry separation (LC-FAIMS-MS) is applied to the untargeted metabolomic analysis of human urine. The time-of-flight mass spectrometer used in the study was scanned at a rate of 20 scans s−1 enabling a FAIMS CF spectrum to be acquired within a 1-s scan time, maintaining an adequate number of data points across each LC peak. The developed method is demonstrated to be able to resolve co-eluting isomeric species and shows good reproducibility (%RSD

Throughout the years an increase has been observed in research output on biochemical markers for determining the postmortem interval (PMI). However, to date, a complete overview is missing on the results of postmortem biochemical markers (PBM''s) for PMI estimation. In this paper, literature was reviewed in order to identify the knowledge lacunae of PBM research from a practical point of view. A three-step approach was undertaken in order to achieve the set goal. Literature was collected, the PBM''s were evaluated for completeness by means of a scorings index based on set criteria, and PBM''s were subsequently evaluated in light of the Daubert & Frye criteria for scientific evidence in court. Seven PBM''s were found to be well investigated, from which potassium had the highest completion score. However, none of these PBM''s could be qualified as suitable for court evidence. Further, this study revealed that the majority of PBM''s (94%) is not well investigated. Consequently, these PBM''s did not meet Daubert & Frye criteria. In order to improve the assessment for use of PBM''s as evidence in court regarding PMI estimation, PBM''s should be investigated more thoroughly and data should be made readily available.

We present here a novel surface mass spectrometry strategy to perform untargeted metabolite profiling of formalin-fixed paraffin-embedded pediatric ependymoma archives. Sequential Orbitrap secondary ion mass spectrometry (3D OrbiSIMS) and liquid extraction surface analysis-tandem mass spectrometry (LESA-MS/MS) permitted the detection of 887 metabolites (163 chemical classes) from pediatric ependymoma tumor tissue microarrays (diameter

Human pluripotent stem cells (hPSCs) can be expanded and differentiatedin vitrointo almost any adult tissue cell type, and thus have great potential as a source for cell therapies with biomedical application. In this study, a fully-defined polymer synthetic substrate is identified for hPSC culture in completely defined, xeno-free conditions. This system can overcome the cost, scalability and reproducibility limitations of current hPSC culture strategies, and facilitate large-scale production. A high-throughput, multi-generational polymer microarray platform approach was used to test over 600 unique polymers and rapidly assess hPSC-polymer interactions in combination with the fully defined xeno-free medium, Essential 8TM(E8). This study identifies as novel nanoscale phase separated blend of poly(tricyclodecane-dimethanol diacrylate) and poly(butyl acrylate) (2:1 v/v), which supports long-term expansion of hPSCs and can be readily coated onto standard cultureware. Analysis of cell-polymer interface interactions through mass spectrometry and integrin blocking studies provides novel mechanistic insight into the role of the E8 proteins in promoting integrin-mediated hPSC attachment and maintaining hPSC signaling, including ability to undergo multi-lineage differentiation. This study therefore identifies a novel substrate for long-term serial passaging of hPSCs in serum-free, commercial chemically-defined E8, which provides a promising and economic hPSC expansion platform for clinical-scale application.

We present here a novel surface mass spectrometry strategy to perform untargeted metabolite profiling of formalin-fixed paraffin-embedded (FFPE) pediatric ependymoma archives. Sequential Orbitrap secondary ion mass spectrometry (3D OrbiSIMS) and liquid extraction surface analysis-tandem MS (LESA-MS/MS) permitted the detection of 887 metabolites (163 chemical classes) from pediatric ependymoma tumor tissue microarrays (diameter Abstract FigureFor Table of Contents Only

Purpose The aim is of this study was to show the poor statistical power of postmortem studies. Further, this study aimed to find an estimate of the effect size for postmortem studies in order to show the importance of this parameter. This can be an aid in performing power analysis to determine a minimal sample size. Methods GPower was used to perform calculations on sample size, effect size, and statistical power. The minimal significance (α) and statistical power (1 − β) were set at 0.05 and 0.80 respectively. Calculations were performed for two groups (Student’s t-distribution) and multiple groups (one-way ANOVA; F-distribution). Results In this study, an average effect size of 0.46 was found (n = 22; SD = 0.30). Using this value to calculate the statistical power of another group of postmortem studies (n = 5) revealed that the average statistical power of these studies was poor (1 − β

A major problem limiting reproducible use of liquid extraction surface analysis (LESA) array sampling of dried surface-deposited liquid samples is the unwanted spread of extraction solvent beyond the dried sample limits, resulting in unreliable data. Here, we explore the use of the Droplet Microarray (DMA), which consists of an array of superhydrophilic spots bordered by a superhydrophobic material giving the potential to confine both the sample spot and the LESA extraction solvent in a defined area. We investigated the DMA method in comparison with a standard glass substrate using LESA analysis of a mixture of biologically relevant compounds with a wide mass range and different physicochemical properties. The optimized DMA method was subsequently applied to urine samples from a human intervention study. Relative standard deviations for the signal intensities were all reduced at least 3-fold when performing LESA-MS on the DMA surface compared with a standard glass surface. Principal component analysis revealed more tight clusters indicating improved spectral reproducibility for a human urine sample extracted from the DMA compared to glass. Lastly, in urine samples from an intervention study, more significant ions (145) were identified when using LESA-MS spectra of control and test urine extracted from the DMA. We demonstrate that DMA provides a surface-assisted LESA-MS method delivering significant improvement of the surface extraction repeatability leading to the acquisition of more robust and higher quality data. The DMA shows potential to be used for LESA-MS for controlled and reproducible surface extraction and for acquisition of high quality, qualitative data in a high-throughput manner.

Recently, Costa et al. published an article about promising biomarkers for estimating the postmortem interval., Recently, Costa et al. published an article about promising biomarkers for estimating the postmortem interval. Instead of postmortem blood, antemortem blood was putrefied in vitro by exposing the blood to a temperature gradient. However, in this way several other influencing factors were excluded, hence, the accuracy of the proposed model is doubtful. Therefore, the aim of this comment is to discuss the methodology, results and shortcomings of the study of Costa et al.