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AnatomyNet: Deep 3D Squeeze-and-excitation U-Nets for fast and fully automated whole-volume anatomical segmentation

PurposeRadiation therapy (RT) is a common treatment for head and neck (HaN) cancer where therapists are often required to manually delineate boundaries of the organs-at-risks (OARs). The radiation therapy planning is time-consuming as each computed tomography (CT) volumetric data set typically consists of hundreds to thousands of slices and needs to be individually inspected. Automated head and neck anatomical segmentation provides a way to speed up and improve the reproducibility of radiation therapy planning. Previous work on anatomical segmentation is primarily based on atlas registrations, which takes up to hours for one patient and requires sophisticated atlas creation. In this work, we propose the AnatomyNet, an end-to-end and atlas-free three dimensional squeeze-and-excitation U-Net (3D SE U-Net), for fast and fully automated whole-volume HaN anatomical segmentation.\n\nMethodsThere are two main challenges for fully automated HaN OARs segmentation: 1) challenge in segmenting small anatomies (i.e., optic chiasm and optic nerves) occupying only a few slices, and 2) training model with inconsistent data annotations with missing ground truth for some anatomical structures because of different RT planning. We propose the AnatomyNet that has one down-sampling layer with the trade-off between GPU memory and feature representation capacity, and 3D SE residual blocks for effective feature learning to alleviate these challenges. Moreover, we design a hybrid loss function with the Dice loss and the focal loss. The Dice loss is a class level distribution loss that depends less on the number of voxels in the anatomy, and the focal loss is designed to deal with highly unbalanced segmentation. For missing annotations, we propose masked loss and weighted loss for accurate and balanced weights updating in the learning of the AnatomyNet.\n\nResultsWe collect 261 HaN CT images to train the AnatomyNet, and use MICCAI Head and Neck Auto Segmentation Challenge 2015 as the benchmark dataset to evaluate the performance of the AnatomyNet. The objective is to segment nine anatomies: brain stem, chiasm, mandible, optic nerve left, optic nerve right, parotid gland left, parotid gland right, submandibular gland left, and submandibular gland right. Compared to previous state-of-the-art methods for each anatomy from the MICCAI 2015 competition, the AnatomyNet increases Dice similarity coefficient (DSC) by 3.3% on average. The proposed AnatomyNet takes only 0.12 seconds on average to segment a whole-volume HaN CT image of an average dimension of 178 x 302 x 225. All the data and code will be availablea.\n\nConclusion1We propose an end-to-end, fast and fully automated deep convolutional network, AnatomyNet, for accurate and whole-volume HaN anatomical segmentation. The proposed Anato-myNet outperforms previous state-of-the-art methods on the benchmark dataset. Extensive experiments demonstrate the effectiveness and good generalization ability of the components in the AnatomyNet.

bioengineering

An algorithm for automated, noninvasive detection of cortical spreading depolarizations based on EEG simulations

We present a novel signal processing algorithm for automated, noninvasive detection of Cortical Spreading Depolarizations (CSDs) using electroencephalography (EEG) signals and validate the algorithm on simulated EEG signals. CSDs are waves of neurochemical changes that suppress neuronal activity as they propagate across the brains cortical surface. CSDs are believed to mediate secondary brain damage after brain trauma and cerebrovascular diseases like stroke. We address key challenges in detecting CSDs from EEG signals: (i) decay of high spatial frequencies as they travel from the cortical surface to the scalp surface; and (ii) presence of sulci and gyri, which makes it difficult to track the CSD waves as they travel across the cortex. Our algorithm detects and tracks \"wavefronts\" of the CSD wave, and stitches together data across space and time to decide on the presence of a CSD wave. To test our algorithm, we provide different models and complex patterns of CSD waves, including different widths of CSD suppressions, and use these models to simulate scalp EEG signals using head models of 4 subjects from the OASIS dataset. Our results suggest that the average width of suppression that a low-density EEG grid of 40 electrodes can detect is 1.1 cm, which includes a vast majority of CSD suppressions, but not all. A higher density EEG grid having 340 electrodes can detect complex CSD patterns as thin as 0.43 cm (less than minimum widths reported in prior works), among which single-gyrus propagation is the hardest to detect because of its small suppression area.

bioengineering

Tendon Structure Quantified using Ultrasound Imaging Differs Based on Location and Training Type

Achilles tendinopathy is ten-times more common amongst running athletes compared to age-matched peers. Load induced tendon remodeling and its progression in an at-risk population of developing symptomatic tendinopathy is not well understood. The purpose of this study was to prospectively characterize Achilles and patellar tendon structure in competitive collegiate distance runners over different competitive seasons using quantitative ultrasound imaging. Twenty-two collegiate cross country runners and eleven controls were examined for this study. Longitudinal and cross-sectional ultrasound images of bilateral Achilles and patellar tendons were obtained at the one week prior to start of formal collegiate cross country practices, one week after the conclusion of cross country season, and one week prior to outdoor track and field championships. Collagen organization, mean echogenicity, tendon thickness, and neovascularity were determined using well established image processing techniques. We found that Achilles and patellar tendons respond differently to high-volume running and transitions from one sport season to another, suggesting that tendon structure is sensitive to differences in tendon loading biomechanics. Our findings indicate that Achilles tendon structure in trained runners differ structurally to control tendons but is stable throughout training while patellar tendon structure changes in response to the transition in training volume between cross country and track seasons. These findings expand upon prior reports that some degree of tendon remodeling may act as a protective adaptation for sport specific loading.\n\nNews and NoteworthyIn this study we prospectively examined the Achilles and patellar tendon structure of distance runners to determine if continued training through multiple seasons elicits tendon remodeling or pathology. We found that Achilles and patellar tendons respond uniquely to the changing loads required during each season. Achilles tendon collagen alignment is mostly stable throughout the competitive cycle, but the patellar tendon undergoes structural changes following the transition from cross-country to track season.

bioengineering

A retrospective evaluation of automated optimization of deep brain stimulation parameters

ObjectiveWe performed a retrospective analysis of an optimization algorithm for the computation of patient-specific multipolar stimulation configurations employing multiple independent current/voltage sources. We evaluated whether the obtained stimulation configurations align with clinical data and whether the optimized stimulation configurations have the potential to lead to an equal or better stimulation of the target region as manual programming, while reducing the time required for programming sessions.\n\nMethodsFor three patients (five electrodes) diagnosed with essential tremor, we derived optimized multipolar stimulation configurations using an approach that is suitable for the application in clinical practice. To evaluate the automatically derived stimulation settings, we compared them to the results of the monopolar review.\n\nResultsWe observe a good agreement between the findings of the monopolar review and the optimized stimulation configurations, with the algorithm assigning the maximal voltage in the optimized multipolar pattern to the contact that was found to lead to the best therapeutic effect in the clinical monopolar review in all cases. Additionally, our simulation results predict that the optimized stimulation settings lead to the activation of an equal or larger volume fraction of the target compared to the manually determined settings in all cases.\n\nConclusionsOur results demonstrate the feasibility of an automatic determination of optimal DBS configurations and motivate a further evaluation of the applied optimization algorithm.

bioengineering

Total lung capacity without plethysmography

BackgroundAmong the most basic measures of respiratory function is the total lung capacity (TLC). TLC is the pulmonary gas volume at maximal lung inflation, which is the sum of the volume of gas that can be exhaled -the vital capacity (VC)- and the volume of gas that cannot -the residual volume (RV). Determination of VC requires only spirometry whereas determination of RV or TLC requires body plethysmography, gas dilution or washout, or thoracic imaging, each of which is more complex than spirometry, and none of which is suited to routine office practice, population screening, or community medicine. To fill this gap, we describe here a new approach to determine TLC without plethysmography.\n\nMethodsIn a heterogeneous population of 434 volunteers (265 male, 169 female; 201 healthy, 170 with airflow obstruction, and 63 with ventilatory restriction), we determined TLC in the standard fashion using conventional body plethysmography (TLCpleth). In the same individuals, we also determined TLC in a novel fashion using the MiniBox (TLCMB). To obtain TLCMB, population-based data from traditional spirometry together with flow-interruption transients were subjected to data mining and machine-learning to create for each individual subject an unbiased statistical determination of TLC.\n\nResultsFor the combined heterogeneous population, we found TLCpleth = 1.02TLCMB -0.091 L, adjusted r2=0.824. For the heterogeneous population as a whole, and for each subpopulation, TLCMB closely tracked TLCpleth. For 26 healthy subjects measured on different days, the coefficient of variation for repeated measurements in was 3.3% for TLCpleth versus 1.6% for TLCMB.\n\nConclusionsThese results establish the validity and potential utility of a new method for rapid, accurate, and repeatable determination of TLC in a heterogeneous patient population, but without the need of a plethysmograph.

bioengineering

Cartilaginous and osteochondral tissue formation by human mesenchymal stem cells on three-dimensionally woven scaffolds

The development of mechanically functional cartilage and bone tissue constructs of clinically relevant size, as well as their integration with native tissues, remain important challenges for regenerative medicine. The objective of this study was to assess adult human mesenchymal stem cells (MSC) in large, three dimensionally woven poly({varepsilon}-caprolactone) (PCL) scaffolds in proximity to viable bone, both in a nude rat subcutaneous pouch model and under simulated conditions in vitro. In Study I, various scaffold permutations: PCL alone, PCL-bone, \"point-of- care\" seeded MSC-PCL-bone, and chondrogenically pre-cultured Ch-MSC-PCL-bone constructs were implanted in a dorsal, ectopic pouch in a nude rat. After eight weeks, only cells in the Ch- MSC-PCL constructs exhibited both chondrogenic and osteogenic gene expression profiles. Notably, while both tissue profiles were present, constructs that had been chondrogenically pre- cultured prior to implantation showed a loss of glycosaminoglycan (GAG) as well as the presence of mineralization along with the formation of trabecula-like structures. In Study II of the study, the GAG loss and mineralization observed in Study I in vivo were recapitulated in vitro by the presence of either nearby bone or osteogenic culture medium additives but were prevented by a continued presence of chondrogenic medium additives. These data suggest conditions under which adult human stem cells in combination with polymer scaffolds synthesize functional and phenotypically distinct tissues based on the environmental conditions, and highlight the potential influence that paracrine factors from adjacent bone may have on MSC fate, once implanted in vivo for chondral or osteochondral repair.

bioengineering

Comparative Multi-scale Hierarchical Structure of the Tail, Plantaris, and Achilles Tendons in the Rat

Rodent tendons are widely used to study human pathology, such as tendinopathy and repair, and to address fundamental physiological questions about development, growth, and remodeling. However, how the gross morphology and the multi-scale hierarchical structure of rat tendons, such as the tail, plantaris, and Achillles tendons, compare to that of human tendons are unknown. In addition, there remains disagreement about terminology and definitions. Specifically, the definition of fascicle and fiber are often dependent on the diameter size and not their characteristic features, which impairs the ability to compare across species where the size of the fiber and fascicle might change with animal size and tendon function. Thus, the objective of the study was to select a single species that is widely used for tendon research (rat) and tendons with varying mechanical functions (tail, plantaris, Achilles) to evaluate the hierarchical structure at multiple length scales. This study was designed including, histology, SEM, and confocal imaging. We confirmed that rat tendons do not contain fascicles, and thus the fiber is the largest tendon subunit in the rat. In addition, we provided a structurally-based definition of a fiber as a bundle of collagen fibrils that is surrounded by elongated cells, and this definition was supported by both histologically processed and unprocessed tendons. In all rat tendons studied, the fiber diameters were consistently 10-50 {micro}m, and this diameter appears to be conserved across larger species. Specific recommendations were made for the strengths and limitations of each rat tendon as tendon research models. Understanding the hierarchical structure of tendon can advance the design and interpretation of experiments and development of tissue engineered constructs.

bioengineering

Survival of selected lactic acid bacteria isolated from pineapple waste in refrigerated pineapple juices

The aim of this research was to isolate and identify lactic acid bacteria (LAB) from pineapple waste. The survival in refrigerated pineapple juices, of a selected isolate with potential probiotic properties, was also studied. The 16S rRNA and pheS partial genes were used to identify LAB, multilocus sequence typing (MLST) genes were used in order to separate strains grouping with Lactobacillus casei and L. paracasei phylogenetically. Tests for survival at pH 2.0, resistance to lysozyme and tolerance to bile salts were used to screen the strains for potential probiotic properties. A L. fermentum isolate was used for the survival study. Three types of pineapple juice made from pulp, a blend of pulp and peel (80:20), and peel extract were inoculated to approximately 106 CFU/mL with L. fermentum and stored at 4 {degrees}C for up to six weeks. The physicochemical composition of juices, including concentration of fermentable sugars and organic acids, total solids content, soluble solids content, titratable acidity and pH, was determined during the survival study. Two genera and five species were identified. Pineapple juices supported the survival of L. fermentum during refrigerated storage but the population of the bacteria decreased over time regardless of the juice type. Juice made from pulp was a more suitable vehicle for the survival of the selected LAB. Some of the juice physicochemical properties, including sugars and organic acids content, pH and titratable acidity, varied significantly (P<0.05) during storage. Further sensory studies are necessary to evaluate consumer acceptance of juices containing the selected isolate.\n\nIMPORTANCELactic acid bacteria (LAB), isolated from pineapple waste, were phylogenetically analyzed and characterized in regards to their tolerance to pH 2.0, lysozyme and bile salts; showing their potential as probiotic strains, if health benefits associated to their ingestion are eventually confirmed. Moreover, pineapple juice supported the survival of Lactobacillus fermentum, isolated from the same food matrix, during refrigerated storage at 4 {degrees}C. Among the three pineapple juices tested (pulp, pulp + peel and peel), L. fermentum survived better in juice made from pulp. However, significant variations were observed overtime in some of the physicochemical properties of the juices including sugars and organic acids content, pH and total titratable acidity.

bioengineering

Gene Correction for SCID-X1 in Long-Term Hematopoietic Stem Cells

Gene correction in human long-term hematopoietic stem cells (LT-HSCs) could be an effective therapy for monogenic diseases of the blood and immune system. High frequencies of reproducible targeted integration of a wild-type cDNA into the endogenous start codon of a gene in LT-HSCs could provide a robust genome editing approach to cure genetic diseases in which patients have different mutations throughout the gene. We describe a clinically relevant method for correcting X-linked severe combined immunodeficiency (SCID-X1). By using a highly specific and active CRISPR/Cas9-AAV6 based strategy and selection-free approach, we achieve up to 20% genome integration frequencies in LT-HSCs of a full-length IL2RG cDNA at the endogenous start site as demonstrated by serial transplantation and analysis of genome edited human cells eight months following initial transplantation. In addition to high frequencies of functional gene correction in LT-HSCs we observed no evidence of abnormal hematopoiesis following transplantation, a functional measure of the lack of genotoxicity. Deep analysis of potential off-target activity detected two sites with low frequency (<0.3%) of off-target mutations. The level of off-target mutations was reduced to below the limit of detection using a high fidelity Cas9. Moreover, karyotype evaluation identified no genomic instability events. We achieved high levels of genome targeting frequencies (median 45%) in CD34+ HSPCs from six SCID-X1 patients and demonstrate rescue of lymphopoietic defect of patient derived cells both in vitro and in vivo. In sum, our study provides specificity, toxicity and efficacy data supportive of clinical development of genome editing to treat SCID-Xl.

bioengineering

Learning 2-in-1: towards integrated EEG-fMRI-neurofeedback

Neurofeedback (NF) allows to exert self-regulation over specific aspects of ones own brain activity by returning information extracted in real-time from brain activity measures. These measures are usually acquired from a single modality, most commonly electroencephalography (EEG) or functional magnetic resonance imaging (fMRI). EEG-fMRI-neurofeedback (EEG-fMRI-NF) is a new approach that consists in providing a NF based simultaneously on EEG and fMRI signals. By exploiting the complementarity of these two modalities, EEG-fMRI-NF opens a new spectrum of possibilities for defining bimodal NF targets that could be more robust, flexible and effective than unimodal ones. Since EEG-fMRI-NF allows for a richer amount of information to be fed back, the question arises of how to represent the EEG and fMRI features simultaneously in order to allow the subject to achieve better self-regulation. In this work, we propose to represent EEG and fMRI features in a single bimodal feedback (integrated feedback). We introduce two integrated feedback strategies for EEG-fMRI-NF and compare their early effects on a motor imagery task with a between-group design. The BiDim group (n=10) was shown a two-dimensional (2D) feedback in which each dimension depicted the information from one modality. The UniDim group (n=10) was shown a one-dimensional (1D) feedback that integrated both types of information even further by merging them into one. Online fMRI activations were significantly higher in the UniDim group than in the BiDim group, which suggests that the 1D feedback is easier to control than the 2D feedback. However subjects from the BiDim group produced more specific BOLD activations with a notably stronger activation in the right superior parietal lobe (BiDim > UniDim, p < 0.001, uncorrected). These results suggest that the 2D feedback encourages subjects to explore their strategies to recruit more specific brain patterns. To summarize, our study shows that 1D and 2D integrated feedbacks are effective but also appear to be complementary and could therefore be used in a bimodal NF training program. Altogether, our study paves the way to novel integrated feedback strategies for the development of flexible and effective bimodal NF paradigms that fully exploits bimodal information and are adapted to clinical applications.

bioengineering

Assessment of abduction movement in older people with painful shoulder: an analysis based on inertial sensors

Reduced range of motion in the shoulder can be a source of functional limitation. Current quantitative evaluation systems are limited to assessing the functionality or the maximum articular amplitudes in each of the planes of movement, both in isolation. These separate clinical evaluation systems may not allow the identification of the underlying impairments contributing to the functional limitation. The use of inertial sensors to quantify movement in addition to more common clinical assessments of the shoulder may allow clinicians to understand that are potentially unnoticed by the human eye. The main objective of this cross-sectional study was to generate an explanatory model for shoulder abduction based on data from inertial sensors. Shoulder abduction of thirteen older adults suffering from shoulder dysfunction was evaluated using two inertial sensors placed on the humerus and scapula. Movement variables (maximum angular mobility, angular peak of velocity, peak of acceleration) were used to explain the functionality of the upper limb (assessed using the Upper Limb Functional Index). Abduction movement of the shoulder was explained by six variables related to the mobility of the shoulder joint complex. A manova analysis was used to explain the results obtained on the functionality of the upper limb. The MANOVA model based on angular mobility explained 69% of the variance of the ULFI value (r-squared=0.69). The most relevant variables were the abduction-adduction of the humerus and the medial and lateral rotation of the scapula. However, given the limited sample size, none of these individual variables were statistically significant in the decomposition model on their own. The method used in the present study reveals the potential importance of the analysis of the scapular and humeral movements for comprehensive evaluation of the upper limb. Further research should include a wider sample and may seek to use this assessment technique in a range of potential clinical applications.

bioengineering

Which osteoarthritic gait features recover following Total Knee Replacement surgery?

BackgroundGait analysis can be used to measure variations in joint function in patients with knee osteoarthritis (OA), and is useful when observing longitudinal biomechanical changes following Total Knee Replacement (TKR) surgery. The Cardiff Classifier is an objective classification tool applied previously to examine the extent of biomechanical recovery following TKR. In this study, it is further developed to reveal the salient features that contribute to recovery towards healthy function.\n\nMethodsGait analysis was performed on 30 patients before and after TKR surgery, and 30 healthy controls. Median TKR follow-up time was 13 months. The combined application of principal component analysis (PCA) and the Cardiff Classifier defined 18 biomechanical features that discriminated OA from healthy gait. Statistical analysis tested whether these features were affected by TKR surgery and, if so, whether they recovered to values found for the controls.\n\nResultsThe Cardiff Classifier successfully discriminated between OA and healthy gait in all 60 cases. Of the 18 discriminatory features, only six (33%) were significantly affected by surgery, including features in all three planes of the ground reaction force (p<0.001), ankle dorsiflexion moment (p<0.001), hip adduction moment (p=0.003), and transverse hip angle (p=0.007). All but two (89%) of these features remained significantly different to those of the control group after surgery.\n\nConclusionsThis approach was able to discriminate gait biomechanics associated with knee OA. The ground reaction force provided the strongest discriminatory features. Despite increased gait velocity and improvements in self-reported pain and function, which would normally be clinical indicators of recovery, the majority of features were not affected by TKR surgery. This TKR cohort retained pre-operative gait patterns; reduced sagittal hip and knee moments, decreased knee flexion, increased hip flexion, and reduced hip adduction. The changes that were associated with surgery were predominantly found at the ankle and hip, rather than at the knee.

bioengineering

Mechanical vibration does not systematically reduce the tremor in Essential Tremor patients

Essential tremor (ET) is a major cause of disability and is not effectively managed in half of the patients. We investigated whether mechanical vibration could reduce tremor in ET by selectively recruiting afferent pathways. We used piezoelectric actuators to deliver vibratory stimuli to the hand and forearm during long trials (4 min), while we monitored the tremor using inertial sensors. We analyzed the effect of four stimulation strategies, including different constant and variable vibration frequencies, in 18 ET patients. Although there was not a clear homogeneous response to vibration across patients and strategies, in most cases (50-72%) mechanical vibration was associated with an increase in the amplitude of their tremor. In contrast, the tremor was reduced in 5-22% of the patients, depending on the strategy. However, these results are hard to interpret given the intrinsic variability of the tremor: during equally long trials without vibration, the tremor changed significantly in 67% of the patients (increased in 45%; decreased in 22%). We conclude that mechanical vibration of the limb does not have a systematic effect on tremor in ET. Moreover, the observed intrinsic variability of the tremor should be taken into account when designing future experiments to assess tremor in ET and how it responds to any intervention.

bioengineering

Restoration of visual function by transplantation of optogenetically engineered photoreceptors

A major challenge in the treatment of retinal degenerative diseases, with the transplantation of replacement photoreceptors, is the difficulty in inducing the grafted cells to grow and maintain light sensitive outer segments (OS) in the host retina, which depends on proper interaction with the underlying retinal pigment epithelium (RPE). For a RPE-independent treatment approach, we introduced a hyperpolarizing microbial opsin into photoreceptor precursors from new-born mice, and transplanted them into blind mice lacking the photoreceptor layer. These optogenetically transformed photoreceptors were light responsive and their transplantation lead to the recovery of visual function, as shown by ganglion cell recordings and behavioral tests. Subsequently, we generated cone photoreceptors from human induced pluripotent stem cells (hiPSCs), expressing the chloride pump Jaws. After transplantation into blind mice, we observed light-driven responses at the photoreceptor and ganglion cell level. These results demonstrate that structural and functional retinal repair is possible by combining stem cell therapy and optogenetics.

bioengineering

ENRICH: a fast method to improve the quality of flexible macromolecular reconstructions

Cryo-electron microscopy using single particle analysis requires the computational averaging of thousands of projection images captured from identical macromolecules. However, macromolecules usually present some degree of flexibility showing different conformations. Computational approaches are then required to classify heterogeneous single particle images into homogeneous sets corresponding to different structural states. Nonetheless, sometimes the attainable resolution of reconstructions obtained from these smaller homogeneous sets is compromised because of reduced number of particles or lack of images at certain macromolecular orientations. In these situations, the current solution to improve map resolution is returning to the electron microscope and collect more data. In this work, we present a fast approach to partially overcome this limitation for heterogeneous data sets. Our method is based on deforming and then moving particles between different conformations using an optical flow approach. Particles are then merged into a unique conformation obtaining reconstructions with improved resolution, contrast and signal-to-noise ratio, then, partially circumventing many issues that impact obtaining high quality reconstructions from small data sets. We present experimental results that show clear improvements in the quality of obtained 3D maps, however, there are also limits to this approach, which we discuss in the manuscript.

bioengineering

Rapid prototyping of a multilayer microphysiological system for primary human intestinal epithelial culture

Here we report benchtop fabrication of multilayer thermoplastic organs-on-chips via laser cut and assembly of double sided adhesives. Biocompatibility was evaluated with Caco-2 cells and primary human intestinal organoids. Chips with Luer fluidic interfaces were economical ($2 per chip) and were fabricated in just hours without use of specialized bonding techniques. Compared with control static Transwell cultures, Caco-2 and organoids cultured on chips formed confluent monolayers expressing tight junctions with low permeability. Caco-2 cells on chip differentiated [~]4 times faster compared to controls and produced mucus. To demonstrate the robustness of laser cut and assembly, we fabricated a dual membrane, tri-layer gut chip integrating 2D monolayers, 3D cell culture, and a basal flow chamber. As proof of concept, we co-cultured a human, differentiated monolayer and intact organoids in a chip with multi-layered contacting compartments. The epithelium exhibited 3D tissue structure and organoids formed in close proximity to the adjacent monolayer. The favorable features of thermoplastics, such as low gas and water vapor permeability, in addition to rapid, facile, and economical fabrication of multilayered devices, make laser cut and assembly an ideal fabrication technique for developing organs-on-chips and studying multicellular tissues.

bioengineering

Ex vivo dynamics of human glioblastoma cells in a microvasculature-on-a-chip system correlates with tumor heterogeneity and subtypes

The perivascular niche (PVN) plays an essential role in brain tumor stem-like cell (BTSC) fate control, tumor invasion, and therapeutic resistance. Herein we report on the use of a microvasculature-on-a-chip system as a PVN model to evaluate the dynamics of BTSCs ex vivo from 10 glioblastoma patients. We observed that BTSCs preferentially localize in the perivascular zone. Live cell tracking revealed that the cells residing in the vicinity of microvessels had the lowest motility, while a fraction of cells on the microvessels unexpectedly possessed the highest motility and migrated over the longest distance. These results indicate that the perivascular zone is a niche for BTSCs, while the microvascular tracks are also a path for long-distance tumor cell migration and invasion. Additionally, the degree of co-localization between tumor cells and microvessels varied significantly across patients. To validate the results from our microvasculature-on-a-chip system, we used single-cell transcriptome sequencing (10 patients and 21,750 single cells in total) to identify the subtype of each tumor cell. The co-localization coefficient was found to correlate positively with proneural (stem-like) or mesenchymal (invasive) but not classical (proliferative) tumor cells. Furthermore, we found that a gene signature profile including PDGFRA correlated strongly with the \"homing\" of brain tumor cells to the PVN. Our findings demonstrated that ex vivo dynamics of human brain tumor cells in a microvasculature-on-a-chip model can recapitulate in vivo tumor cell dynamics, heterogeneity, and subtypes, representing a new route to the study of human tumor cell biology and uncover patient-specific tumor cell functions.

bioengineering

Rapid energy expenditure estimation for assisted and inclined loaded walking

BackgroundEstimating energy expenditure with indirect calorimetry requires expensive equipment and provides slow and noisy measurements. Rapid estimates using wearable sensors would enable techniques like optimizing assistive devices outside a lab. Existing methods correlate data from wearable sensors to measured energy expenditure without evaluating the accuracy of the estimated energy expenditure for activity conditions or subjects not included in the correlation process. Our goal is to assess data-driven models that are capable of rapidly estimating energy expenditure for new conditions and subjects.\n\nMethodsWe developed models that estimated energy expenditure from two datasets during walking conditions with (1) ankle exoskeleton assistance and (2) various loads and inclines. The estimation was portable and rapid, using input features that are possible to measure with wearable sensors and restricting the input data length to a single gait cycle or four second interval. The performance of the models was evaluated for three use cases. The first case estimated energy expenditure during walking conditions for subjects with some subject specific training data available. The second case estimated all conditions in the dataset for a new subject not included in the training data. The third case estimated new conditions for a new subject. The models also ordered the magnitude of energy expenditure across all conditions for a new subject.\n\nResultsThe average errors in energy expenditure estimation during assisted walking conditions were 4.4%, 8.0%, and 8.1% for the three use cases, respectively. The average errors in energy expenditure estimation during inclined and loaded walking conditions were 6.1%, 9.7%, and 11.7% for the three use cases. The models ordered the magnitude of energy expenditure with a maximum and average percentage of correctly ordered conditions of 56% and 43% for assisted walking and 85% and 55% for incline and loaded walking.\n\nConclusionsOur data-driven models determined the accuracy of energy expenditure estimation for three use cases. For experiments where the accuracy of a data-driven model is sufficient, standard indirect calorimetry can be replaced. The energy expenditure ordering could aid in selecting optimal assistance conditions. The models, code, and datasets are provided for reproduction and extension of our results.

bioengineering