Tracking induced pluripotent stem cell differentiation with a fluorescent genetically encoded epigenetic probe.

Epigenetic modifications (methylation, acetylation, etc.) of core histones play a key role in regulation of gene expression. Thus, the epigenome changes strongly during various biological processes such as cell differentiation and dedifferentiation. Classical methods of analysis of epigenetic modifications such as mass-spectrometry and chromatin immuno-precipitation, work with fixed cells only. Here we present a genetically encoded fluorescent probe, MPP8-Green, for detecting H3K9me3, a histone modification associated with inactive chromatin. This probe, based on the chromodomain of MPP8, allows for visualization of H3K9me3 epigenetic landscapes in single living cells. We used this probe to track changes in H3K9me3 landscapes during the differentiation of induced pluripotent stem cells (iPSCs) into induced neurons. Our findings revealed two major waves of global H3K9me3 reorganization during 4-day differentiation, namely on the first and third days, whereas nearly no changes occurred on the second and fourth days. The proposed method LiveMIEL (Live-cell Microscopic Imaging of Epigenetic Landscapes), which combines genetically encoded epigenetic probes and machine learning approaches, enables classification of multiparametric epigenetic signatures of single cells during stem cell differentiation and potentially in other biological models.

Epistemic uncertainty challenges aging clock reliability in predicting rejuvenation effects.

Epigenetic aging clocks have been widely used to validate rejuvenation effects during cellular reprogramming. However, these predictions are unverifiable because the true biological age of reprogrammed cells remains unknown. We present an analytical framework to consider rejuvenation predictions from the uncertainty perspective. Our analysis reveals that the DNA methylation profiles across reprogramming are poorly represented in the aging data used to train clock models, thus introducing high epistemic uncertainty in age estimations. Moreover, predictions of different published clocks are inconsistent, with some even suggesting zero or negative rejuvenation. While not questioning the possibility of age reversal, we show that the high clock uncertainty challenges the reliability of rejuvenation effects observed during in vitro reprogramming before pluripotency and throughout embryogenesis. Conversely, our method reveals a significant age increase after in vivo reprogramming. We recommend including uncertainty estimation in future aging clock models to avoid the risk of misinterpreting the results of biological age prediction.

Deep reinforcement learning with significant multiplications inference.

We propose a sparse computation method for optimizing the inference of neural networks in reinforcement learning (RL) tasks. Motivated by the processing abilities of the brain, this method combines simple neural network pruning with a delta-network algorithm to account for the input data correlations. The former mimics neuroplasticity by eliminating inefficient connections; the latter makes it possible to update neuron states only when their changes exceed a certain threshold. This combination significantly reduces the number of multiplications during the neural network inference for fast neuromorphic computing. We tested the approach in popular deep RL tasks, yielding up to a 100-fold reduction in the number of required multiplications without substantial performance loss (sometimes, the performance even improved).

Optimal MRI undersampling patterns for ultimate benefit of medical vision tasks.

Compressed sensing is commonly concerned with optimizing the image quality after a partial undersampling of the measurable k-space to accelerate MRI. In this article, we propose to change the focus from the quality of the reconstructed image to the quality of the downstream image analysis outcome. Specifically, we propose to optimize the patterns according to how well a sought-after pathology could be detected or localized in the reconstructed images. We find the optimal undersampling patterns in k-space that maximize target value functions of interest in commonplace medical vision problems (reconstruction, segmentation, and classification) and propose a new iterative gradient sampling routine universally suitable for these tasks. We validate the proposed MRI acceleration paradigm on three classical medical datasets, demonstrating a noticeable improvement of the target metrics at the high acceleration factors (for the segmentation problem at ×16 acceleration, we report up to 12% improvement in Dice score over the other undersampling patterns).

Medical image captioning via generative pretrained transformers.

The proposed model for automatic clinical image caption generation combines the analysis of radiological scans with structured patient information from the textual records. It uses two language models, the Show-Attend-Tell and the GPT-3, to generate comprehensive and descriptive radiology records. The generated textual summary contains essential information about pathologies found, their location, along with the 2D heatmaps that localize each pathology on the scans. The model has been tested on two medical datasets, the Open-I, MIMIC-CXR, and the general-purpose MS-COCO, and the results measured with natural language assessment metrics demonstrated its efficient applicability to chest X-ray image captioning.

Evaluation of an Integrated Spectroscopy and Classification Platform for Point-of-Care Core Needle Biopsy Assessment: Performance Characteristics from Ex Vivo Renal Mass Biopsies.

PURPOSE: To evaluate a transmission optical spectroscopy instrument for rapid ex vivo assessment of core needle cancer biopsies (CNBs) at the point of care. MATERIALS AND METHODS: CNBs from surgically resected renal tumors and nontumor regions were scanned on their sampling trays with a custom spectroscopy instrument. After extracting principal spectral components, machine learning was used to train logistic regression, support vector machines, and random decision forest (RF) classifiers on 80% of randomized and stratified data. The algorithms were evaluated on the remaining 20% of the data set held out during training. Binary classification (tumor/nontumor) was performed based on a decision threshold. Multinomial classification was also performed to differentiate between the subtypes of renal cell carcinoma (RCC) and account for potential confounding effects from fat, blood, and necrotic tissue. Classifiers were compared based on sensitivity, specificity, and positive predictive value (PPV) relative to a histopathologic standard. RESULTS: A total of 545 CNBs from 102 patients were analyzed, yielding 5,583 spectra after outlier exclusion. At the individual spectra level, the best performing algorithm was RF with sensitivities of 96% and 92% and specificities of 90% and 89%, for the binary and multiclass analyses, respectively. At the full CNB level, RF algorithm also showed the highest sensitivity and specificity (93% and 91%, respectively). For RCC subtypes, the highest sensitivity and PPV were attained for clear cell (93.5%) and chromophobe (98.2%) subtypes, respectively. CONCLUSIONS: Ex vivo spectroscopy imaging paired with machine learning can accurately characterize renal mass CNB at the time of tissue acquisition.

Infrared Laser-Based Single Cell Permeabilization by Plasma Membrane Temperature Gradients.

Single cell microinjection provides precise tuning of the volume and timing of delivery into the treated cells; however, it also introduces workflow complexity that requires highly skilled operators and specialized equipment. Laser-based microinjection provides an alternative method for targeting a single cell using a common laser and a workflow that may be readily standardized. This paper presents experiments using a 1550 nm, 100 fs pulse duration laser with a repetition rate of 20 ns for laser-based microinjection and calculations of the hypothesized physical mechanism responsible for the experimentally observed permeabilization. Chinese Hamster Ovarian (CHO) cells exposed to this laser underwent propidium iodide uptake, demonstrating the potential for selective cell permeabilization. The agreement between the experimental conditions and the electropermeabilization threshold based on estimated changes in the transmembrane potential induced by a laser-induced plasma membrane temperature gradient, even without accounting for enhancement due to traditional electroporation, strengthens the hypothesis of this mechanism for the experimental observations. Compared to standard 800 nm lasers, 1550 nm fs lasers may ultimately provide a lower cost microinjection method that readily interfaces with a microscope and is agnostic to operator skill, while inducing fewer deleterious effects (e.g., temperature rise, shockwaves, and cavitation bubbles).

MOOD 2020: A Public Benchmark for Out-of-Distribution Detection and Localization on Medical Images.

Detecting Out-of-Distribution (OoD) data is one of the greatest challenges in safe and robust deployment of machine learning algorithms in medicine. When the algorithms encounter cases that deviate from the distribution of the training data, they often produce incorrect and over-confident predictions. OoD detection algorithms aim to catch erroneous predictions in advance by analysing the data distribution and detecting potential instances of failure. Moreover, flagging OoD cases may support human readers in identifying incidental findings. Due to the increased interest in OoD algorithms, benchmarks for different domains have recently been established. In the medical imaging domain, for which reliable predictions are often essential, an open benchmark has been missing. We introduce the Medical-Out-Of-Distribution-Analysis-Challenge (MOOD) as an open, fair, and unbiased benchmark for OoD methods in the medical imaging domain. The analysis of the submitted algorithms shows that performance has a strong positive correlation with the perceived difficulty, and that all algorithms show a high variance for different anomalies, making it yet hard to recommend them for clinical practice. We also see a strong correlation between challenge ranking and performance on a simple toy test set, indicating that this might be a valuable addition as a proxy dataset during anomaly detection algorithm development.

Tubular shape aware data generation for segmentation in medical imaging.

PURPOSE: Chest X-ray is one of the most widespread examinations of the human body. In interventional radiology, its use is frequently associated with the need to visualize various tube-like objects, such as puncture needles, guiding sheaths, wires, and catheters. Detection and precise localization of these tube-like objects in the X-ray images are, therefore, of utmost value, catalyzing the development of accurate target-specific segmentation algorithms. Similar to the other medical imaging tasks, the manual pixel-wise annotation of the tubes is a resource-consuming process. METHODS: In this work, we aim to alleviate the lack of annotated images by using artificial data. Specifically, we present an approach for synthetic generation of the tube-shaped objects, with a generative adversarial network being regularized with a prior-shape constraint. Namely, our model uses Frangi-based regularization to draw synthetic tubes in the predefined fake mask regions and, then, uses the adversarial component to preserve the global realistic appearance of the synthesized image. RESULTS: Our method eliminates the need for the paired image-mask data and requires only a weakly labeled dataset, with fine-tuning on a small paired sample (10-20 images) proving sufficient to reach the accuracy of the fully supervised models. CONCLUSION: We report the applicability of the approach for the task of segmenting tubes and catheters in the X-ray images, whereas the results should also hold for the other acquisition modalities and image computing applications that contain tubular objects.

3D denoised completion network for deep single-pixel reconstruction of hyperspectral images.

Single-pixel imaging acquires an image by measuring its coefficients in a transform domain, thanks to a spatial light modulator. However, as measurements are sequential, only a few coefficients can be measured in the real-time applications. Therefore, single-pixel reconstruction is usually an underdetermined inverse problem that requires regularization to obtain an appropriate solution. Combined with a spectral detector, the concept of single-pixel imaging allows for hyperspectral imaging. While each channel can be reconstructed independently, we propose to exploit the spectral redundancy between channels to regularize the reconstruction problem. In particular, we introduce a denoised completion network that includes 3D convolution filters. Contrary to black-box approaches, our network combines the classical Tikhonov theory with the deep learning methodology, leading to an explainable network. Considering both simulated and experimental data, we demonstrate that the proposed approach yields hyperspectral images with higher quantitative metrics than the approaches developed for grayscale images.