Platform-independent estimation of human physiological time from single blood samples.

Abundant epidemiological evidence links circadian rhythms to human health, from heart disease to neurodegeneration. Accurate determination of an individual's circadian phase is critical for precision diagnostics and personalized timing of therapeutic interventions. To date, however, we still lack an assay for physiological time that is accurate, minimally burdensome to the patient, and readily generalizable to new data. Here, we present TimeMachine, an algorithm to predict the human circadian phase using gene expression in peripheral blood mononuclear cells from a single blood draw. Once trained on data from a single study, we validated the trained predictor against four independent datasets with distinct experimental protocols and assay platforms, demonstrating that it can be applied generalizably. Importantly, TimeMachine predicted circadian time with a median absolute error ranging from 1.65 to 2.7 h, regardless of systematic differences in experimental protocol and assay platform, without renormalizing the data or retraining the predictor. This feature enables it to be flexibly applied to both new samples and existing data without limitations on the transcriptomic profiling technology (microarray, RNAseq). We benchmark TimeMachine against competing approaches and identify the algorithmic features that contribute to its performance.

Feature-specific quantile normalization and feature-specific mean-variance normalization deliver robust bi-directional classification and feature selection performance between microarray and RNAseq data.

BACKGROUND: Cross-platform normalization seeks to minimize technological bias between microarray and RNAseq whole-transcriptome data. Incorporating multiple gene expression platforms permits external validation of experimental findings, and augments training sets for machine learning models. Here, we compare the performance of Feature Specific Quantile Normalization (FSQN) to a previously used but unvalidated and uncharacterized method we label as Feature Specific Mean Variance Normalization (FSMVN). We evaluate the performance of these methods for bidirectional normalization in the context of nested feature selection. RESULTS: FSQN and FSMVN provided clinically equivalent bidirectional model performance with and without feature selection for colon CMS and breast PAM50 classification. Using principal component analysis, we determine that these methods eliminate batch effects related to technological platforms. Without feature selection, no statistical difference was identified between the performance of FSQN and FSMVN of cross-platform data compared to within-platform distributions. Under optimal feature selection conditions, balanced accuracy was FSQN and FSMVN were statistically equivalent to the within-platform distribution performance in multivariable linear regression analysis. FSQN and FSMVN also provided similar performance to within-platform distributions as the number of selected genes used to create models decreases. CONCLUSIONS: In the context of generating supervised machine learning classifiers for molecular subtypes, FSQN and FSMVN are equally effective. Under optimal modeling conditions, FSQN and FSMVN provide equivalent model accuracy performance on cross-platform normalization data compared to within-platform data. Using cross-platform data should still be approached with caution as subtle performance differences may exist depending on the classification problem, training, and testing distributions.

Individualized detection of TMPRSS2-ERG fusion status in prostate cancer: a rank-based qualitative transcriptome signature.

BACKGROUND: TMPRSS2-ERG (T2E) fusion is highly related to aggressive clinical features in prostate cancer (PC), which guides individual therapy. However, current fusion prediction tools lacked enough accuracy and biomarkers were unable to be applied to individuals across different platforms due to their quantitative nature. This study aims to identify a transcriptome signature to detect the T2E fusion status of PC at the individual level. METHODS: Based on 272 high-throughput mRNA expression profiles from the Sboner dataset, we developed a rank-based algorithm to identify a qualitative signature to detect T2E fusion in PC. The signature was validated in 1223 samples from three external datasets (Setlur, Clarissa, and TCGA). RESULTS: A signature, composed of five mRNAs coupled to ERG (five ERG-mRNA pairs, 5-ERG-mRPs), was developed to distinguish T2E fusion status in PC. 5-ERG-mRPs reached 84.56% accuracy in Sboner dataset, which was verified in Setlur dataset (n = 455, accuracy = 82.20%) and Clarissa dataset (n = 118, accuracy = 81.36%). Besides, for 495 samples from TCGA, two subtypes classified by 5-ERG-mRPs showed a higher level of significance in various T2E fusion features than subtypes obtained through current fusion prediction tools, such as STAR-Fusion. CONCLUSIONS: Overall, 5-ERG-mRPs can robustly detect T2E fusion in PC at the individual level, which can be used on any gene measurement platform without specific normalization procedures. Hence, 5-ERG-mRPs may serve as an auxiliary tool for PC patient management.

Cross-platform comparison of immune signatures in immunotherapy-treated patients with advanced melanoma using a rank-based scoring approach.

BACKGROUND: Gene expression profiling is increasingly being utilised as a diagnostic, prognostic and predictive tool for managing cancer patients. Single-sample scoring approach has been developed to alleviate instability of signature scores due to variations from sample composition. However, it is a challenge to achieve comparable signature scores across different expressional platforms. METHODS: The pre-treatment biopsies from a total of 158 patients, who have received single-agent anti-PD-1 (n = 84) or anti-PD-1 + anti-CTLA-4 therapy (n = 74), were performed using NanoString PanCancer IO360 Panel. Multiple immune-related signature scores were measured from a single-sample rank-based scoring approach, singscore. We assessed the reproducibility and the performance in reporting immune profile of singscore based on NanoString assay in advance melanoma. To conduct cross-platform analyses, singscores between the immune profiles of NanoString assay and the previous orthogonal whole transcriptome sequencing (WTS) data were compared through linear regression and cross-platform prediction. RESULTS: singscore-derived signature scores reported significantly high scores in responders in multiple PD-1, MHC-1-, CD8 T-cell-, antigen presentation-, cytokine- and chemokine-related signatures. We found that singscore provided stable and reproducible signature scores among the repeats in different batches and cross-sample normalisations. The cross-platform comparisons confirmed that singscores derived via NanoString and WTS were comparable. When singscore of WTS generated by the overlapping genes to the NanoString gene set, the signatures generated highly correlated cross-platform scores (Spearman correlation interquartile range (IQR) [0.88, 0.92] and r2 IQR [0.77, 0.81]) and better prediction on cross-platform response (AUC = 86.3%). The model suggested that Tumour Inflammation Signature (TIS) and Personalised Immunotherapy Platform (PIP) PD-1 are informative signatures for predicting immunotherapy-response outcomes in advanced melanoma patients treated with anti-PD-1-based therapies. CONCLUSIONS: Overall, the outcome of this study confirms that singscore based on NanoString data is a feasible approach to produce reliable signature scores for determining patients' immune profiles and the potential clinical utility in biomarker implementation, as well as to conduct cross-platform comparisons, such as WTS.

CrossICC: iterative consensus clustering of cross-platform gene expression data without adjusting batch effect.

Unsupervised clustering of high-throughput gene expression data is widely adopted for cancer subtyping. However, cancer subtypes derived from a single dataset are usually not applicable across multiple datasets from different platforms. Merging different datasets is necessary to determine accurate and applicable cancer subtypes but is still embarrassing due to the batch effect. CrossICC is an R package designed for the unsupervised clustering of gene expression data from multiple datasets/platforms without the requirement of batch effect adjustment. CrossICC utilizes an iterative strategy to derive the optimal gene signature and cluster numbers from a consensus similarity matrix generated by consensus clustering. This package also provides abundant functions to visualize the identified subtypes and evaluate subtyping performance. We expected that CrossICC could be used to discover the robust cancer subtypes with significant translational implications in personalized care for cancer patients. AVAILABILITY AND IMPLEMENTATION: The package is implemented in R and available at GitHub (https://github.com/bioinformatist/CrossICC) and Bioconductor (http://bioconductor.org/packages/release/bioc/html/CrossICC.html) under the GPL v3 License.

Molecular Evolutionary Genetics Analysis (MEGA) for macOS.

The Molecular Evolutionary Genetics Analysis (MEGA) software enables comparative analysis of molecular sequences in phylogenetics and evolutionary medicine. Here, we introduce the macOS version of the MEGA software. This new version eliminates the need for virtualization and emulation programs previously required to use MEGA on Apple computers. MEGA for macOS utilizes memory and computing resources efficiently for conducting evolutionary analyses on macOS. It has a native Cocoa graphical user interface that is programmed to provide a consistent user experience across macOS, Windows, and Linux. MEGA for macOS is available from www.megasoftware.net free of charge.

Universal method for robust detection of circadian state from gene expression.

Circadian clocks play a key role in regulating a vast array of biological processes, with significant implications for human health. Accurate assessment of physiological time using transcriptional biomarkers found in human blood can significantly improve diagnosis of circadian disorders and optimize the delivery time of therapeutic treatments. To be useful, such a test must be accurate, minimally burdensome to the patient, and readily generalizable to new data. A major obstacle in development of gene expression biomarker tests is the diversity of measurement platforms and the inherent variability of the data, often resulting in predictors that perform well in the original datasets but cannot be universally applied to new samples collected in other settings. Here, we introduce TimeSignature, an algorithm that robustly infers circadian time from gene expression. We demonstrate its application in data from three independent studies using distinct microarrays and further validate it against a new set of samples profiled by RNA-sequencing. Our results show that TimeSignature is more accurate and efficient than competing methods, estimating circadian time to within 2 h for the majority of samples. Importantly, we demonstrate that once trained on data from a single study, the resulting predictor can be universally applied to yield highly accurate results in new data from other studies independent of differences in study population, patient protocol, or assay platform without renormalizing the data or retraining. This feature is unique among expression-based predictors and addresses a major challenge in the development of generalizable, clinically useful tests.

Content and Dynamics of Websites Shared Over Vaccine-Related Tweets in COVID-19 Conversations: Computational Analysis.

BACKGROUND: The onset of the COVID-19 pandemic and the consequent "infodemic" increased concerns about Twitter's role in advancing antivaccination messages, even before a vaccine became available to the public. New computational methods allow for analysis of cross-platform use by tracking links to websites shared over Twitter, which, in turn, can uncover some of the content and dynamics of information sources and agenda-setting processes. Such understanding can advance theory and efforts to reduce misinformation. OBJECTIVE: Informed by agenda-setting theory, this study aimed to identify the content and temporal patterns of websites shared in vaccine-related tweets posted to COVID-19 conversations on Twitter between February and June 2020. METHODS: We used triangulation of data analysis methods. Data mining consisted of the screening of around 5 million tweets posted to COVID-19 conversations to identify tweets that related to vaccination and including links to websites shared within these tweets. We further analyzed the content the 20 most-shared external websites using a mixed methods approach. RESULTS: Of 841,896 vaccination-related tweets identified, 185,994 (22.1%) contained links to specific websites. A wide range of websites were shared, with the 20 most-tweeted websites constituting 14.5% (27,060/185,994) of the shared websites and typically being shared for only 2 to 3 days. Traditional media constituted the majority of these 20 websites, along with other social media and governmental sources. We identified markers of inauthentic propagation for some of these links. CONCLUSIONS: The topic of vaccination was prevalent in tweets about COVID-19 early in the pandemic. Sharing websites was a common communication strategy, and its "bursty" pattern and inauthentic propagation strategies pose challenges for health promotion efforts. Future studies should consider cross-platform use in dissemination of health information and in counteracting misinformation.

SELAMAT: A New Secure and Lightweight Multi-Factor Authentication Scheme for Cross-Platform Industrial IoT Systems.

The development of the industrial Internet of Things (IIoT) promotes the integration of the cross-platform systems in fog computing, which enable users to obtain access to multiple application located in different geographical locations. Fog users at the network's edge communicate with many fog servers in different fogs and newly joined servers that they had never contacted before. This communication complexity brings enormous security challenges and potential vulnerability to malicious threats. The attacker may replace the edge device with a fake one and authenticate it as a legitimate device. Therefore, to prevent unauthorized users from accessing fog servers, we propose a new secure and lightweight multi-factor authentication scheme for cross-platform IoT systems (SELAMAT). The proposed scheme extends the Kerberos workflow and utilizes the AES-ECC algorithm for efficient encryption keys management and secure communication between the edge nodes and fog node servers to establish secure mutual authentication. The scheme was tested for its security analysis using the formal security verification under the widely accepted AVISPA tool. We proved our scheme using Burrows Abdi Needham's logic (BAN logic) to prove secure mutual authentication. The results show that the SELAMAT scheme provides better security, functionality, communication, and computation cost than the existing schemes.

A large-scale comparative study of isoform expressions measured on four platforms.

BACKGROUND: Most eukaryotic genes produce different transcripts of multiple isoforms by inclusion or exclusion of particular exons. The isoforms of a gene often play diverse functional roles, and thus it is necessary to accurately measure isoform expressions as well as gene expressions. While previous studies have demonstrated the strong agreement between mRNA sequencing (RNA-seq) and array-based gene and/or isoform quantification platforms (Microarray gene expression and Exon-array), the more recently developed NanoString platform has not been systematically evaluated and compared, especially in large-scale studies across different cancer domains. RESULTS: In this paper, we present a large-scale comparative study among RNA-seq, NanoString, array-based, and RT-qPCR platforms using 46 cancer cell lines across different cancer types. The goal is to understand and evaluate the calibers of the platforms for measuring gene and isoform expressions in cancer studies. We first performed NanoString experiments on 59 cancer cell lines with 404 custom-designed probes for measuring the expressions of 478 isoforms in 155 genes, and additional RT-qPCR experiments for a subset of the measured isoforms in 13 cell lines. We then combined the data with the matched RNA-seq, Exon-array, and Microarray data of 46 of the 59 cell lines for the comparative analysis. CONCLUSION: In the comparisons of the platforms for measuring the expressions at both isoform and gene levels, we found that (1) the agreement on isoform expressions is lower than the agreement on gene expressions across the four platforms; (2) NanoString and Exon-array are not consistent on isoform quantification even though both techniques are based on hybridization reactions; (3) RT-qPCR experiments are more consistent with RNA-seq and Exon-array than NanoString in isoform quantification; (4) different RNA-seq isoform quantification methods show varying estimation results, and among the methods, Net-RSTQ and eXpress are more consistent across the platforms; and (5) RNA-seq has the best overall consistency with the other platforms on gene expression quantification.

Facilitating the development of cross-platform mHealth applications for chronic supportive care and a case study.

Mobile health (mHealth) apps have received increasing attention, due to their abilities to support patients who suffer from various conditions. mHealth apps may be especially helpful for patients with chronic diseases, by providing pertinent information, tracking symptoms, and inspiring adherence to medication regimens. To achieve these objectives, researchers need to prototype mHealth apps with dedicated software architectures. In this paper, a cloud-based mHealth application development concept is presented for chronic patient supportive care apps. The concept integrates existing software platforms and services for simplified app development that can be reused for other target applications. This developmental method also facilitates app portability, through the use of common components found across multiple mobile platforms, and scalability, through the loose coupling of services. The results are demonstrated by the development of native Android and cross-platform web apps, in a case study that presents an mHealth solution for endocrine hormone therapy (EHT). A performance analysis methodology, an app usability evaluation, based on focus group responses, and alpha and pre-beta testing results are provided.

Comparative evaluation of isoform-level gene expression estimation algorithms for RNA-seq and exon-array platforms.

Given that the majority of multi-exon genes generate diverse functional products, it is important to evaluate expression at the isoform level. Previous studies have demonstrated strong gene-level correlations between RNA sequencing (RNA-seq) and microarray platforms, but have not studied their concordance at the isoform level. We performed transcript abundance estimation on raw RNA-seq and exon-array expression profiles available for common glioblastoma multiforme samples from The Cancer Genome Atlas using different analysis pipelines, and compared both the isoform- and gene-level expression estimates between programs and platforms. The results showed better concordance between RNA-seq/exon-array and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) platforms for fold change estimates than for raw abundance estimates, suggesting that fold change normalization against a control is an important step for integrating expression data across platforms. Based on RT-qPCR validations, eXpress and Multi-Mapping Bayesian Gene eXpression (MMBGX) programs achieved the best performance for RNA-seq and exon-array platforms, respectively, for deriving the isoform-level fold change values. While eXpress achieved the highest correlation with the RT-qPCR and exon-array (MMBGX) results overall, RSEM was more highly correlated with MMBGX for the subset of transcripts that are highly variable across the samples. eXpress appears to be most successful in discriminating lowly expressed transcripts, but IsoformEx and RSEM correlate more strongly with MMBGX for highly expressed transcripts. The results also reinforce how potentially important isoform-level expression changes can be masked by gene-level estimates, and demonstrate that exon arrays yield comparable results to RNA-seq for evaluating isoform-level expression changes.

SpinStudioJ: A cross-platform NMR data acquisition and processing workbench based on a plug-in architecture.

Flexibility and extensibility are important issues in the design of nuclear magnetic resonance (NMR) software, as these determine the ability to integrate a variety of continuously evolving data acquisition and processing methods. Here, SpinStudioJ is introduced. It is an NMR data acquisition and processing workbench with a plug-in-based architecture. The workbench is based on Eclipse Rich Client Platform, which provides a plug-and-play runtime mechanism and rich graphical user interface functionality. New data acquisition methods and processing algorithms can be easily integrated into the SpinStudioJ workbench by defining extension points, without the need to redistribute existing modules. The software is independent of operating systems, as it leverages the cross-platform feature of the Java virtual machine.

Animations in Cross-Platform Mobile Applications: An Evaluation of Tools, Metrics and Performance.

Along with the proliferation of high-end and performant mobile devices, we find that the inclusion of visually animated user interfaces are commonplace, but that research on their performance is scarce. Thus, for this study, eight mobile apps have been developed for scrutiny and assessment to report on the device hardware impact and penalties caused by transitions and animations, with an emphasis on apps generated using cross-platform development frameworks. The tasks we employ for animation performance measuring, are those of (i) a complex animation consisting of multiple elements, (ii) the opening sequence of a side menu navigation pattern, and (iii) a transition animation during in-app page navigation. We employ multiple performance profiling tools, and scrutinize metrics including frames per second (FPS), CPU usage, device memory usage and GPU memory usage, all to uncover the impact caused by executing transitions and animations. We uncover important differences in device hardware utilization during animations across the different cross-platform technologies employed. Additionally, Android and iOS are found to differ greatly in terms of memory consumption, CPU usage and rendered FPS, a discrepancy that is true for both the native and cross-platform apps. The findings we report are indeed factors contributing to the complexity of app development.

A new cross-platform architecture for epi-info software suite.

BACKGROUND: The Epi-Info software suite, built and maintained by the Centers for Disease Control and Prevention (CDC), is widely used by epidemiologists and public health researchers to collect and analyze public health data, especially in the event of outbreaks such as Ebola and Zika. As it exists today, Epi-Info Desktop runs only on the Windows platform, and the larger Epi-Info Suite of products consists of separate codebases for several different devices and use-cases. Software portability has become increasingly important over the past few years as it offers a number of obvious benefits. These include reduced development time, reduced cost, and simplified system architecture. Thus, there is a blatant need for continued research. Specifically, it is critical to fully understand any underlying negative performance issues which arise from platform-agnostic systems. Such understanding should allow for improved design, and thus result in substantial mitigation of reduced performance. In this paper, we present a viable cross-platform architecture for Epi-Info which solves many of these problems. RESULTS: We have successfully generated executables for Linux, Mac, and Windows from a single code-base, and we have shown that performance need not be completely sacrificed when building a cross-platform application. This has been accomplished by using Electron as a wrapper for an AngularJS app, a Python analytics module, and a local, browser-based NoSQL database. CONCLUSIONS: Promising results warrant future research. Specifically, the design allows for cross-platform form-design, data-collection, offline/online modes, scalable storage, automatic local-to-remote data sync, and fast analytics which rival more traditional approaches.

Dual-labelled antibodies for flow and mass cytometry: A new tool for cross-platform comparison and enrichment of target cells for mass cytometry.

Antibody conjugates applicable in both conventional flow and mass cytometry would offer interesting options for cross-platform comparison, as well as the enrichment of rare target cells by conventional flow cytometry (FC) sorting prior to deep phenotyping by mass cytometry (MC). Here, we introduce a simple method to generate dual fluorochrome/metal-labelled antibodies by consecutive orthogonal labelling. First, we compared different fluorochrome-conjugated antibodies specific for CD4, such as FITC, Vio667, VioGreen or VioBlue for their compatibility with the conventional secondary MAXPAR® labelling protocol. After labelling with 141 Pr, the fluorescence emission spectra of all fluorochromes investigated retained their characteristics, and CD4 dual conjugates (DCs) provided consistent results in immune phenotyping assays performed by FC and MC. The phenotypical composition of CD4+ T-cells was maintained after enrichment by FC sorting using different CD4 DCs. Finally, magnetic cell depletion was combined with FC sorting using CD19-VioBlue-142 Nd, CD20-VioGreen-147 Sm, CD27-Cy5-167 Er and CD38-Alexa488-143 Nd DC to enrich rare human plasmablasts to purities >80%, which allowed a subsequent deep phenotyping by MC. In conclusion, DCs have been successfully established for direct assay comparison between FC and MC, and help to minimise MC data acquisition time for deep phenotyping of rare cell subsets.

A decision support system to follow up and diagnose primary headache patients using semantically enriched data.

BACKGROUND: Headache disorders are an important health burden, having a large health-economic impact worldwide. Current treatment & follow-up processes are often archaic, creating opportunities for computer-aided and decision support systems to increase their efficiency. Existing systems are mostly completely data-driven, and the underlying models are a black-box, deteriorating interpretability and transparency, which are key factors in order to be deployed in a clinical setting. METHODS: In this paper, a decision support system is proposed, composed of three components: (i) a cross-platform mobile application to capture the required data from patients to formulate a diagnosis, (ii) an automated diagnosis support module that generates an interpretable decision tree, based on data semantically annotated with expert knowledge, in order to support physicians in formulating the correct diagnosis and (iii) a web application such that the physician can efficiently interpret captured data and learned insights by means of visualizations. RESULTS: We show that decision tree induction techniques achieve competitive accuracy rates, compared to other black- and white-box techniques, on a publicly available dataset, referred to as migbase. Migbase contains aggregated information of headache attacks from 849 patients. Each sample is labeled with one of three possible primary headache disorders. We demonstrate that we are able to reduce the classification error, statistically significant (ρ≤0.05), with more than 10% by balancing the dataset using prior expert knowledge. Furthermore, we achieve high accuracy rates by using features extracted using the Weisfeiler-Lehman kernel, which is completely unsupervised. This makes it an ideal approach to solve a potential cold start problem. CONCLUSION: Decision trees are the perfect candidate for the automated diagnosis support module. They achieve predictive performances competitive to other techniques on the migbase dataset and are, foremost, completely interpretable. Moreover, the incorporation of prior knowledge increases both predictive performance as well as transparency of the resulting predictive model on the studied dataset.

Mediator lipidomics by liquid chromatography-tandem mass spectrometry.

Comprehensive quantitative analysis of lipid mediators using liquid chromatography-tandem mass spectrometry is an effective strategy in the elucidation of disease mechanisms; but technically, it has been and is still a great challenge to achieve reliable datasets that cover variety of lipid metabolites contained at trace levels in complex biological matrices. In this opinion article, we introduce our experiences in developing lipid mediator profiling systems, and deliver some comments on limitations of current methodology.

Evaluation of Smartphone Inertial Sensor Performance for Cross-Platform Mobile Applications.

Smartphone sensors are being increasingly used in mobile applications. The performance of sensors varies considerably among different smartphone models and the development of a cross-platform mobile application might be a very complex and demanding task. A publicly accessible resource containing real-life-situation smartphone sensor parameters could be of great help for cross-platform developers. To address this issue we have designed and implemented a pilot participatory sensing application for measuring, gathering, and analyzing smartphone sensor parameters. We start with smartphone accelerometer and gyroscope bias and noise parameters. The application database presently includes sensor parameters of more than 60 different smartphone models of different platforms. It is a modest, but important start, offering information on several statistical parameters of the measured smartphone sensors and insights into their performance. The next step, a large-scale cloud-based version of the application, is already planned. The large database of smartphone sensor parameters may prove particularly useful for cross-platform developers. It may also be interesting for individual participants who would be able to check-up and compare their smartphone sensors against a large number of similar or identical models.

A set of external reference controls/probes that enable quality assurance between different microarray platforms.

RNA external standards, although important to ensure equivalence across many microarray platforms, have yet to be fully implemented in the research community. In this article, a set of unique RNA external standards (or RNA standards) and probe pairs that were added to total RNA in the samples before amplification and labeling are described. Concentration-response curves of RNA external standards were used across multiple commercial DNA microarray platforms and/or quantitative real-time polymerase chain reaction (RT-PCR) and next-generation sequencing to identify problematic assays and potential sources of variation in the analytical process. A variety of standards can be added in a range of concentrations spanning high and low abundances, thereby enabling the evaluation of assay performance across the expected range of concentrations found in a clinical sample. Using this approach, we show that we are able to confirm the dynamic range and the limit of detection for each DNA microarray platform, RT-PCR protocol, and next-generation sequencer. In addition, the combination of a series of standards and their probes was investigated on each platform, demonstrating that multiplatform calibration and validation is possible.