Identifying novel regulatory effects for clinically relevant genes through the study of the Greek population.

BACKGROUND: Expression quantitative trait loci (eQTL) studies provide insights into regulatory mechanisms underlying disease risk. Expanding studies of gene regulation to underexplored populations and to medically relevant tissues offers potential to reveal yet unknown regulatory variants and to better understand disease mechanisms. Here, we performed eQTL mapping in subcutaneous (S) and visceral (V) adipose tissue from 106 Greek individuals (Greek Metabolic study, GM) and compared our findings to those from the Genotype-Tissue Expression (GTEx) resource. RESULTS: We identified 1,930 and 1,515 eGenes in S and V respectively, over 13% of which are not observed in GTEx adipose tissue, and that do not arise due to different ancestry. We report additional context-specific regulatory effects in genes of clinical interest (e.g. oncogene ST7) and in genes regulating responses to environmental stimuli (e.g. MIR21, SNX33). We suggest that a fraction of the reported differences across populations is due to environmental effects on gene expression, driving context-specific eQTLs, and suggest that environmental effects can determine the penetrance of disease variants thus shaping disease risk. We report that over half of GM eQTLs colocalize with GWAS SNPs and of these colocalizations 41% are not detected in GTEx. We also highlight the clinical relevance of S adipose tissue by revealing that inflammatory processes are upregulated in individuals with obesity, not only in V, but also in S tissue. CONCLUSIONS: By focusing on an understudied population, our results provide further candidate genes for investigation regarding their role in adipose tissue biology and their contribution to disease risk and pathogenesis.

Combining Multiple RNA-Seq Data Analysis Algorithms Using Machine Learning Improves Differential Isoform Expression Analysis.

RNA sequencing has become the standard technique for high resolution genome-wide monitoring of gene expression. As such, it often comprises the first step towards understanding complex molecular mechanisms driving various phenotypes, spanning organ development to disease genesis, monitoring and progression. An advantage of RNA sequencing is its ability to capture complex transcriptomic events such as alternative splicing which results in alternate isoform abundance. At the same time, this advantage remains algorithmically and computationally challenging, especially with the emergence of even higher resolution technologies such as single-cell RNA sequencing. Although several algorithms have been proposed for the effective detection of differential isoform expression from RNA-Seq data, no widely accepted golden standards have been established. This fact is further compounded by the significant differences in the output of different algorithms when applied on the same data. In addition, many of the proposed algorithms remain scarce and poorly maintained. Driven by these challenges, we developed a novel integrative approach that effectively combines the most widely used algorithms for differential transcript and isoform analysis using state-of-the-art machine learning techniques. We demonstrate its usability by applying it on simulated data based on several organisms, and using several performance metrics; we conclude that our strategy outperforms the application of the individual algorithms. Finally, our approach is implemented as an R Shiny application, with the underlying data analysis pipelines also available as docker containers.

Ten simple rules for making training materials FAIR.

Everything we do today is becoming more and more reliant on the use of computers. The field of biology is no exception; but most biologists receive little or no formal preparation for the increasingly computational aspects of their discipline. In consequence, informal training courses are often needed to plug the gaps; and the demand for such training is growing worldwide. To meet this demand, some training programs are being expanded, and new ones are being developed. Key to both scenarios is the creation of new course materials. Rather than starting from scratch, however, it's sometimes possible to repurpose materials that already exist. Yet finding suitable materials online can be difficult: They're often widely scattered across the internet or hidden in their home institutions, with no systematic way to find them. This is a common problem for all digital objects. The scientific community has attempted to address this issue by developing a set of rules (which have been called the Findable, Accessible, Interoperable and Reusable [FAIR] principles) to make such objects more findable and reusable. Here, we show how to apply these rules to help make training materials easier to find, (re)use, and adapt, for the benefit of all.

Exome Sequencing in BRCA1- and BRCA2-Negative Greek Families Identifies MDM1 and NBEAL1 as Candidate Risk Genes for Hereditary Breast Cancer.

Approximately 10% of breast cancer (BC) cases are hereditary BC (HBC), with HBC most commonly encountered in the context of hereditary breast and ovarian cancer (HBOC) syndrome. Although thousands of loss-of-function (LoF) alleles in over 20 genes have been associated with HBC susceptibility, the genetic etiology of approximately 50% of cases remains unexplained, even when polygenic risk models are considered. We focused on one of the least-studied European populations and applied whole-exome sequencing (WES) to 52 individuals from 17 Greek HBOC families, in which at least one patient was negative for known HBC risk variants. Initial screening revealed pathogenic variants in known cancer genes, including BARD1:p.Trp91* detected in a cancer-free individual, and MEN1:p.Glu260Lys detected in a BC patient. Gene- and variant-based approaches were applied to exome data to identify candidate risk variants outside of known risk genes. Findings were verified in a collection of Canadian HBOC patients of European ancestry (FBRCAX), in an independent group of Canadian BC patients (CHUM-BC) and controls (CARTaGENE), as well as in individuals from The Cancer Genome Atlas (TCGA) and the UK Biobank (UKB). Rare LoF variants were uncovered in MDM1 and NBEAL1 in Greek and Canadian HBOC patients. We also report prioritized missense variants SETBP1:c.4129G > C and C7orf34:c.248C > T. These variants comprise promising candidates whose role in cancer pathogenicity needs to be explored further.

Sociodemographic Indicators of Health Status Using a Machine Learning Approach and Data from the English Longitudinal Study of Aging (ELSA).

BACKGROUND Studies on the effects of sociodemographic factors on health in aging now include the use of statistical models and machine learning. The aim of this study was to evaluate the determinants of health in aging using machine learning methods and to compare the accuracy with traditional methods. MATERIAL AND METHODS The health status of 6,209 adults, age <65 years (n=1,585), 65-79 years (n=3,267), and >80 years (n=1,357) were measured using an established health metric (0-100) that incorporated physical function and activities of daily living (ADL). Data from the English Longitudinal Study of Ageing (ELSA) included socio-economic and sociodemographic characteristics and history of falls. Health-trend and personal-fitted variables were generated as predictors of health metrics using three machine learning methods, random forest (RF), deep learning (DL) and the linear model (LM), with calculation of the percentage increase in mean square error (%IncMSE) as a measure of the importance of a given predictive variable, when the variable was removed from the model. RESULTS Health-trend, physical activity, and personal-fitted variables were the main predictors of health, with the%incMSE of 85.76%, 63.40%, and 46.71%, respectively. Age, employment status, alcohol consumption, and household income had the%incMSE of 20.40%, 20.10%, 16.94%, and 13.61%, respectively. Performance of the RF method was similar to the traditional LM (p=0.7), but RF significantly outperformed DL (p=0.006). CONCLUSIONS Machine learning methods can be used to evaluate multidimensional longitudinal health data and may provide accurate results with fewer requirements when compared with traditional statistical modeling.

Machine learning methodologies versus cardiovascular risk scores, in predicting disease risk.

BACKGROUND: The use of Cardiovascular Disease (CVD) risk estimation scores in primary prevention has long been established. However, their performance still remains a matter of concern. The aim of this study was to explore the potential of using ML methodologies on CVD prediction, especially compared to established risk tool, the HellenicSCORE. METHODS: Data from the ATTICA prospective study (n = 2020 adults), enrolled during 2001-02 and followed-up in 2011-12 were used. Three different machine-learning classifiers (k-NN, random forest, and decision tree) were trained and evaluated against 10-year CVD incidence, in comparison with the HellenicSCORE tool (a calibration of the ESC SCORE). Training datasets, consisting from 16 variables to only 5 variables, were chosen, with or without bootstrapping, in an attempt to achieve the best overall performance for the machine learning classifiers. RESULTS: Depending on the classifier and the training dataset the outcome varied in efficiency but was comparable between the two methodological approaches. In particular, the HellenicSCORE showed accuracy 85%, specificity 20%, sensitivity 97%, positive predictive value 87%, and negative predictive value 58%, whereas for the machine learning methodologies, accuracy ranged from 65 to 84%, specificity from 46 to 56%, sensitivity from 67 to 89%, positive predictive value from 89 to 91%, and negative predictive value from 24 to 45%; random forest gave the best results, while the k-NN gave the poorest results. CONCLUSIONS: The alternative approach of machine learning classification produced results comparable to that of risk prediction scores and, thus, it can be used as a method of CVD prediction, taking into consideration the advantages that machine learning methodologies may offer.

A comparison of statistical and machine-learning techniques in evaluating the association between dietary patterns and 10-year cardiometabolic risk (2002-2012): the ATTICA study.

Statistical methods are usually applied in examining diet-disease associations, whereas factor analysis is commonly used for dietary pattern recognition. Recently, machine learning (ML) has been also proposed as an alternative technique in health classification. In this work, the predictive accuracy of statistical v. ML methodologies as regards the association of dietary patterns on CVD risk was tested. During 2001-2002, 3042 men and women (45 (sd 14) years) were enrolled in the ATTICA study. In 2011-2012, the 10-year CVD follow-up was performed among 2020 participants. Item Response Theory was applied to create a metric of combined 10-year cardiometabolic risk, the 'Cardiometabolic Health Score', that incorporated incidence of CVD, diabetes, hypertension and hypercholesterolaemia. Factor analysis was performed to extract dietary patterns, on the basis of either foods or nutrients consumed; linear regression analysis was used to assess their association with the cardiometabolic score. Two ML techniques (k-nearest-neighbor's algorithm and random-forests decision tree) were applied to evaluate participants' health based on dietary information. Factor analysis revealed five and three factors from foods and nutrients, respectively, explaining 54 and 65 % of the total variation in intake. Nutrient and food pattern regression models showed similar accuracy in correctly classifying an individual according to the cardiometabolic risk (R 2=9·6 % and R 2=8·3 %, respectively). ML techniques were superior compared with linear regression in correct classification of the individuals according to the Health Score (accuracy approximately 38 v. 6 %, respectively), whereas the two ML methods showed equal classification ability. Conclusively, ML methods could be a valuable tool in the field of nutritional epidemiology, leading to more accurate disease-risk evaluation.

Comparison of block matching and differential methods for motion analysis of the carotid artery wall from ultrasound images.

Motion of the carotid artery wall is important for the quantification of arterial elasticity and contractility and can be estimated with a number of techniques. In this paper, a framework for quantitative evaluation of motion analysis techniques from B-mode ultrasound images is introduced. Six synthetic sequences were produced using 1) a real image corrupted by Gaussian and speckle noise of 25 and 15 dB, and 2) the ultrasound simulation package Field II. In both cases, a mathematical model was used, which simulated the motion of the arterial wall layers and the surrounding tissue, in the radial and longitudinal directions. The performance of four techniques, namely optical flow (OF (HS)), weighted least-squares optical flow (OF (LK(WLS))), block matching (BM), and affine block motion model (ABMM), was investigated in the context of this framework. The average warping indices were lowest for OF (LK(WLS)) (1.75 pixels), slightly higher for ABMM (2.01 pixels), and highest for BM (6.57 pixels) and OF (HS) (11.57 pixels). Due to its superior performance, OF (LK(WLS)) was used to quantify motion of selected regions of the arterial wall in real ultrasound image sequences of the carotid artery. Preliminary results indicate that OF (LK(WLS)) is promising, because it efficiently quantified radial, longitudinal, and shear strains in healthy adults and diseased subjects.

Detection and isolation of antiatherogenic and antioxidant substances present in olive mill wastes by a novel filtration system.

Olive mill waste water (OMWW) is a major environmental issue in the Mediterranean. We address this problem by investigating the wastes for the presence of biologically active compounds already detected in both olive oil and pomace. Two initial OMWW samples were filtered using two microporous filtering media: (a) clayey diatomite and (b) zeolitic volcanic tuffs, obtaining three filtered samples from each. All initial and filtrated samples were tested for their activity on platelet activating factor (PAF)-induced aggregation. The results showed that the initial samples contain biologically active compounds (PAF inhibitors) and that in their respective last-eluted filtered samples these compounds are purified. These eluted samples, along with their corresponding initial OMWW, were further separated with HPLC and the purified fractions responsible for the aforementioned biological activity, were further studied using chemical determinations and MS analysis. It was confirmed that the PAF inhibitor present in these fractions resembles the one isolated from olive oil. These results offer a new approach on the OMWW handling by offering an alternative use of this waste as starting material for nutritional and/or pharmaceutical purposes in the future.