Label-free cell classification in holographic flow cytometry through an unbiased learning strategy.

Nowadays, label-free imaging flow cytometry at the single-cell level is considered the stepforward lab-on-a-chip technology to address challenges in clinical diagnostics, biology, life sciences and healthcare. In this framework, digital holography in microscopy promises to be a powerful imaging modality thanks to its multi-refocusing and label-free quantitative phase imaging capabilities, along with the encoding of the highest information content within the imaged samples. Moreover, the recent achievements of new data analysis tools for cell classification based on deep/machine learning, combined with holographic imaging, are urging these systems toward the effective implementation of point of care devices. However, the generalization capabilities of learning-based models may be limited from biases caused by data obtained from other holographic imaging settings and/or different processing approaches. In this paper, we propose a combination of a Mask R-CNN to detect the cells, a convolutional auto-encoder, used to the image feature extraction and operating on unlabelled data, thus overcoming the bias due to data coming from different experimental settings, and a feedforward neural network for single cell classification, that operates on the above extracted features. We demonstrate the proposed approach in the challenging classification task related to the identification of drug-resistant endometrial cancer cells.

An automated pipeline integrating AlphaFold 2 and MODELLER for protein structure prediction.

The ability to predict a protein's three-dimensional conformation represents a crucial starting point for investigating evolutionary connections with other members of the corresponding protein family, examining interactions with other proteins, and potentially utilizing this knowledge for the purpose of rational drug design. In this work, we evaluated the feasibility of improving AlphaFold2's three-dimensional protein predictions by developing a novel pipeline (AlphaMod) that incorporates AlphaFold2 with MODELLER, a template-based modeling program. Additionally, our tool can drive a comprehensive quality assessment of the tertiary protein structure by incorporating and comparing a set of different quality assessment tools. The outcomes of selected tools are combined into a composite score (BORDASCORE) that exhibits a meaningful correlation with GDT_TS and facilitates the selection of optimal models in the absence of a reference structure. To validate AlphaMod's results, we conducted evaluations using two distinct datasets summing up to 72 targets, previously used to independently assess AlphaFold2's performance. The generated models underwent evaluation through two methods: i) averaging the GDT_TS scores across all produced structures for a single target sequence, and ii) a pairwise comparison of the best structures generated by AlphaFold2 and AlphaMod. The latter, within the unsupervised setups, shows a rising accuracy of approximately 34% over AlphaFold2. While, when considering the supervised setup, AlphaMod surpasses AlphaFold2 in 18% of the instances. Finally, there is an 11% correspondence in outcomes between the diverse methodologies. Consequently, AlphaMod's best-predicted tertiary structures in several cases exhibited a significant improvement in the accuracy of the predictions with respect to the best models obtained by AlphaFold2. This pipeline paves the way for the integration of additional data and AI-based algorithms to further improve the reliability of the predictions.

Machine Learning as a Support for the Diagnosis of Type 2 Diabetes.

Diabetes is a chronic, metabolic disease characterized by high blood sugar levels. Among the main types of diabetes, type 2 is the most common. Early diagnosis and treatment can prevent or delay the onset of complications. Previous studies examined the application of machine learning techniques for prediction of the pathology, and here an artificial neural network shows very promising results as a possible valuable aid in the management and prevention of diabetes. Additionally, its superior ability for long-term predictions makes it an ideal choice for this field of study. We utilized machine learning methods to uncover previously undiscovered associations between an individual's health status and the development of type 2 diabetes, with the goal of accurately predicting its onset or determining the individual's risk level. Our study employed a binary classifier, trained on scratch, to identify potential nonlinear relationships between the onset of type 2 diabetes and a set of parameters obtained from patient measurements. Three datasets were utilized, i.e., the National Center for Health Statistics' (NHANES) biennial survey, MIMIC-III and MIMIC-IV. These datasets were then combined to create a single dataset with the same number of individuals with and without type 2 diabetes. Since the dataset was balanced, the primary evaluation metric for the model was accuracy. The outcomes of this study were encouraging, with the model achieving accuracy levels of up to 86% and a ROC AUC value of 0.934. Further investigation is needed to improve the reliability of the model by considering multiple measurements from the same patient over time.

StaSiS-Net: A stacked and siamese disparity estimation network for depth reconstruction in modern 3D laparoscopy.

Developing accurate and real-time algorithms for a non-invasive three-dimensional representation and reconstruction of internal patient structures is one of the main research fields in computer-assisted surgery and endoscopy. Mono and stereo endoscopic images of soft tissues are converted into a three-dimensional representation by the estimation of depth maps. However, automatic, detailed, accurate and robust depth map estimation is a challenging problem that, in the stereo setting, is strictly dependent on a robust estimate of the disparity map. Many traditional algorithms are often inefficient or not accurate. In this work, novel self-supervised stacked and Siamese encoder/decoder neural networks are proposed to compute accurate disparity maps for 3D laparoscopy depth estimation. These networks run in real-time on standard GPU-equipped desktop computers and the outputs may be used for depth map estimation using the a known camera calibration. We compare performance on three different public datasets and on a new challenging simulated dataset and our solutions outperform state-of-the-art mono and stereo depth estimation methods. Extensive robustness and sensitivity analyses on more than 30000 frames has been performed. This work leads to important improvements in mono and stereo real-time depth map estimation of soft tissues and organs with a very low average mean absolute disparity reconstruction error with respect to ground truth.

Blind microscopy image denoising with a deep residual and multiscale encoder/decoder network.

In computer-aided diagnosis (CAD) focused on microscopy, denoising improves the quality of image analysis. In general, the accuracy of this process may depend both on the experience of the microscopist and on the equipment sensitivity and specificity. A medical image could be corrupted by several perturbations during image acquisition. Nowadays, CAD deep learning applications pre-process images with image denoising models to reinforce learning and prediction. In this work, an innovative and lightweight deep multiscale convolutional encoder-decoder neural network is proposed. Specifically, the encoder uses deterministic mapping to map features into a hidden representation. Then, the latent representation is rebuilt to generate the reconstructed denoised image. Residual learning strategies are used to improve and accelerate the training process using skip connections in bridging across convolutional and deconvolutional layers. The proposed model reaches on average 38.38 of PSNR and 0.98 of SSIM on a test set of 57458 images overcoming state-of-the-art models in the same application domain.Clinical relevance - Encoder-decoder based denoiser enables industry experts to provide more accurate and reliable medical interpretation and diagnosis in a variety of fields, from microscopy to surgery, with the benefit of real-time processing.

Signal metrics analysis of oscillatory patterns in bacterial multi-omic networks.

MOTIVATION: One of the branches of Systems Biology is focused on a deep understanding of underlying regulatory networks through the analysis of the biomolecules oscillations and their interplay. Synthetic Biology exploits gene or/and protein regulatory networks towards the design of oscillatory networks for producing useful compounds. Therefore, at different levels of application and for different purposes, the study of biomolecular oscillations can lead to different clues about the mechanisms underlying living cells. It is known that network-level interactions involve more than one type of biomolecule as well as biological processes operating at multiple omic levels. Combining network/pathway-level information with genetic information it is possible to describe well-understood or unknown bacterial mechanisms and organism-specific dynamics. RESULTS: Following the methodologies used in signal processing and communication engineering, a methodology is introduced to identify and quantify the extent of multi-omic oscillations. These are due to the process of multi-omic integration and depend on the gene positions on the chromosome. Ad hoc signal metrics are designed to allow further biotechnological explanations and provide important clues about the oscillatory nature of the pathways and their regulatory circuits. Our algorithms designed for the analysis of multi-omic signals are tested and validated on 11 different bacteria for thousands of multi-omic signals perturbed at the network level by different experimental conditions. Information on the order of genes, codon usage, gene expression and protein molecular weight is integrated at three different functional levels. Oscillations show interesting evidence that network-level multi-omic signals present a synchronized response to perturbations and evolutionary relations along taxa. AVAILABILITY AND IMPLEMENTATION: The algorithms, the code (in language R), the tool, the pipeline and the whole dataset of multi-omic signal metrics are available at: https://github.com/lodeguns/Multi-omicSignals. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

A study on multi-omic oscillations in Escherichia coli metabolic networks.

BACKGROUND: Two important challenges in the analysis of molecular biology information are data (multi-omic information) integration and the detection of patterns across large scale molecular networks and sequences. They are are actually coupled beause the integration of omic information may provide better means to detect multi-omic patterns that could reveal multi-scale or emerging properties at the phenotype levels. RESULTS: Here we address the problem of integrating various types of molecular information (a large collection of gene expression and sequence data, codon usage and protein abundances) to analyse the E.coli metabolic response to treatments at the whole network level. Our algorithm, MORA (Multi-omic relations adjacency) is able to detect patterns which may represent metabolic network motifs at pathway and supra pathway levels which could hint at some functional role. We provide a description and insights on the algorithm by testing it on a large database of responses to antibiotics. Along with the algorithm MORA, a novel model for the analysis of oscillating multi-omics has been proposed. Interestingly, the resulting analysis suggests that some motifs reveal recurring oscillating or position variation patterns on multi-omics metabolic networks. Our framework, implemented in R, provides effective and friendly means to design intervention scenarios on real data. By analysing how multi-omics data build up multi-scale phenotypes, the software allows to compare and test metabolic models, design new pathways or redesign existing metabolic pathways and validate in silico metabolic models using nearby species. CONCLUSIONS: The integration of multi-omic data reveals that E.coli multi-omic metabolic networks contain position dependent and recurring patterns which could provide clues of long range correlations in the bacterial genome.