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A dual-fluorescence assay for gene delivery vehicle screening in macrophages with an inflammation-inducible reporter construct

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BMC methods 2025年第1期2卷 8页

作者： Allie Ivy Shelby N Bess Shilpi Agrawal Varun Kochar Abbey L Stokes Timothy J Muldoon Christopher E Nelson Department of Biomedical Engineering University of Arkansas 120 John A. White Jr. Engineering Hall Fayetteville AR 72701 USA. Cell and Molecular Biology Program University of Arkansas Fayetteville AR USA.

Background:Macrophages are a promising target for therapeutics in various applications such as regenerative medicine and immunotherapy for cancer. Due to their plastic nature, macrophages can switch from a non-activated state to activated with the smallest environmental change. For macrophages to be effective in their respective applications, screening for phenotypic changes is necessary to elucidate the cell response to different delivery vehicles, vaccines, small molecules, and other stimuli. Methods:We created a sensitive and dynamic high-throughput screening method for macrophages based on the activation of NF-κB. For this reporter, we placed an mRFP1 fluorescence gene under the control of an inflammatory promoter, which recruits NF-κB response elements to promote expression during the inflammatory response in macrophages. We characterized the inflammatory reporter based on key markers of an inflammatory response in macrophages including TNF-α cytokine release and immunostaining for inflammatory and non-inflammatory cell surface markers. We compared gene delivery and inflammation of several clinically relevant viral vehicles and commercially available non-viral vehicles. Statistical analysis between groups was performed with a one-way ANOVA with post-hoc Tukey's test. Results:The reporter macrophages demonstrated a dynamic range after LPS stimulation with an EC50 of 0.61 ng/mL that was highly predictive of TNF-α release. Flow cytometry revealed heterogeneity between groups but confirmed population level shifts in pro-inflammatory markers. Finally, we demonstrated utility of the reporter by showing divergent effects with various leading gene delivery vehicles. Discussion:This screening technique developed here provides a dynamic, high-throughput screening technique for determining inflammatory response by mouse macrophages to specific stimuli. The method presented here provides insight into the inflammatory response in mouse macrophages to different viral and non

关键词： Gene delivery Inflammation Macrophage NF-κB Reporter

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Convergent effects of peptides on the initiation of feeding motor programs in the mollusk Aplysia

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Journal of Neurophysiology 2025年第5期133卷 1368-1379页

作者： Evans, Colin G. Barry, Michael A. Reaver, Carrie N. Patel, Paras R. Chestek, Cynthia A. Perkins, Matthew H. Jing, Jian Cropper, Elizabeth C. Department of Neuroscience Friedman Brain Institute Icahn School of Medicine at Mount Sinai New York NY United States Department of Biomedical Engineering Biointerfaces Institute University of Michigan Ann Arbor MI United States Department of Electrical Engineering and Computer Science Neurosciences Program Robotics Program University of Michigan Ann Arbor MI United States State Key Laboratory of Pharmaceutical Biotechnology Institute for Brain Sciences Chemistry and Biomedicine Innovation Center Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnology Advanced Institute for Life Sciences School of Life Sciences Nanjing University Nanjing China

Neuropeptides configure the feeding network of Aplysia. For example, egestive activity is promoted by small cardioactive peptide (SCP), and ingestive activity is promoted by a combination of feeding circuit activating peptide (FCAP) and cerebral peptide 2 (CP-2). In addition, SCP and FCAP/CP-2 have a common network effect that does not contribute to motor program specification. They increase the excitability of an interneuron, B63. In this report, we further characterized the effects of peptides on B63. We performed voltage-clamp experiments and used a step protocol to look at steady-state currents. We found that SCP and FCAP/CP-2 both induced an inward current that was virtually absent in low-sodium saline. Previous work has established that B63 is unusual in the feeding circuit in that subthreshold depolarizations are autonomously generated that can trigger motor programs. Here, we show that this autonomous activity is more frequent in the presence of peptides. Previous studies have also shown that activity of the feeding central pattern generator (CPG) can be initiated by neurons that excite B63, e.g., by cerebral buccal interneuron 2 (CBI-2), a projection neuron that triggers biting-like motor programs. Here, we show that the latency of CBI-2-induced activity is decreased by stimulation of the esophageal nerve (EN) (which releases endogenous SCP). These results, taken together with previous results, indicate that peptides that act divergently to configure network activity additionally act convergently to promote motor program induction. We present data that suggest that this arrangement facilitates brief switches between ingestive and egestive motor activity. © 2025 The Authors.

关键词： central pattern generator invertebrate modulation motor program neuropeptide

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Evaluating Crosslinking Efficiency and Cytocompatibility of Three Commonly Used Photoinitiators Across Different Cell-Compatible Hydrogel Platforms

SSRN

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SSRN 2025年

作者： Gavish Moscovitz, Aya Simaan Yameen, Haneen Bar-Am, Orit Seliktar, Dror The Faculty of Biomedical Engineering Technion-Israel Institute of Technology Haifa Israel The Interdisciplinary Program in Biotechnology Technion–Israel Institute of Technology Haifa Israel

biomedical hydrogels that undergo a rapid liquid-to-solid phase transition often use a photopolymerization strategy to crosslink the polymer network. In applications such as 3D bioprinting or tissue engineering that involve cell-laden hydrogels, or where in-vivo polymerization is desired (i.e., in-situ), cytotoxicity of the hydrogel crosslinking reaction becomes an important issue. There are only a few cytocompatible photoinitiators (PIs) that are commonly used for crosslinking biomedical hydrogels, including Irgacure2959 (I2959), lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) and EosinY (EY). These PIs are illuminated at 365nm, 405nm and 525nm, respectively. The photopolymerization conditions, including PI concentration and illumination intensity may affect both the cytotoxicity of the reaction and the crosslinking of the hydrogel. Herein we tested these three PIs to optimize the maximum crosslinking efficiency while minimizing cell death associated with the photochemical reaction. Testing was performed on three different types of cell-compatible hydrogels used for cell encapsulation and 3D cell growth, including a synthetic material (poly(ethylene glycol)-diacrylate, PEG-DA), a semi-synthetic material (PEG-fibrinogen, PF) and a modified biological material (methacrylated fibrinogen, FibMA). Strain-rate controlled rheology was used to characterize the crosslinking reaction efficiency, both in terms of maximum shear storage modulus, G’, and crosslinking kinetics. Viability was examined on human dermal fibroblasts in 3D culture within photopolymerized PF hydrogels crosslinked using each of the PIs. The results showed that PI concentration and the illumination intensity had significant impact on the crosslinking efficiency, with each material system demonstrating different responses to the PI parameters. Optimal photo-crosslinking conditions, considering both crosslinking efficiency and cell viability were not the same for the modified protein hydrogels, as co

关键词： Cell death

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Using baseline MRI radiomic features to predict the efficacy of repetitive transcranial magnetic stimulation in Alzheimer’s patients

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Medical and Biological engineering and Computing 2025年 1-11页

作者： Saha, Chandan Figley, Chase R. Lithgow, Brian Wang, Xikui Fitzgerald, Paul B. Koski, Lisa Mansouri, Behzad Moussavi, Zahra Biomedical Engineering Program University of Manitoba Winnipeg Canada Department of Radiology and Biomedical Engineering Program University of Manitoba Winnipeg Canada Riverview Health Center Winnipeg Canada Warren Center for Actuarial Studies and Research University of Manitoba Winnipeg Canada School of Medicine and Psychology Australian National University Canberra Australia Department of Neurology & Neurosurgery McGill University Montreal Canada Brain Vision and Concussion Clinic-iScope Winnipeg Canada

The efficacy of repetitive transcranial magnetic stimulation (rTMS) as a treatment for Alzheimer’s disease (AD) is uncertain at baseline. Herein, we aimed to investigate whether radiomic features from the pre-treatment MRI data could predict rTMS efficacy for AD treatment. Out of 110 participants with AD in the active (n = 75) and sham (n = 35) rTMS treatment groups having T1-weighted brain MRI data, we had two groups of responders (active = 55 and sham = 24) and non-responders (active = 20 and sham = 11). We extracted histogram-based radiomic features from MRI data using 3D Slicer software;the most important features were selected utilizing a combination of a two-sample t-test, correlation test, least absolute shrinkage, and selection operator. The support vector machine classified rTMS responders and non-responders with a cross-validated mean accuracy/AUC of 81.9%/90.0% in the active group and 87.4%/95.8% in the sham group. Further, the radiomic features of the active group significantly correlated with participants’ AD assessment scale-cognitive subscale (ADAS-Cog) change after treatment (false discovery rate corrected p © International Federation for Medical and Biological engineering 2025.

关键词： Neurodegenerative diseases

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Reconfiguring Surface Acoustic Wave Microfluidics via In Situ Control of Elastic Wave Polarization

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Physical Review Letters 2025年第3期134卷 037002-037002页

作者： Yu Gao (郜煜) Thomas Voglhuber-Brunnmaier Yuekang Li Leyla Akh Nicholas H. Patino Apresio Kefin Fajrial Massimo Ruzzene Bernhard Jakoby Xiaoyun Ding Paul M. Rady Department of Mechanical Engineering University of Colorado Boulder Colorado 80309 USA Institute for Microelectronics and Microsensors Johannes Kepler University Linz Austria Biomedical Engineering Program University of Colorado Boulder Colorado 80309 USA Materials Science and Engineering Program University of Colorado Boulder Colorado 80309 USA BioFrontiers Institute University of Colorado Boulder Colorado 80309 USA

We demonstrate in situ control of the elastic wave polarization in a surface acoustic wave (SAW). It allows us to create highly reconfigurable SAW microfluidics that can be switched on demand between the acoustohydrodynamic (AHD) regime and electrohydrodynamic (EHD) regime for manipulating particles and cells. The control of wave polarization comes from our experimental and theoretical identification of an unexpected shear-horizontal (SH) wave mode in a conventional Rayleigh (R) wave design, which is stereotyped to excite only vertically polarized Rayleigh SAWs. The SH wave mode is predominantly horizontally polarized and can be selectively excited to propagate in the same direction as the Rayleigh SAW. Such a selective wave generation between the SH mode and R mode allows for reconfiguration between AHD and EHD regimes that leads to unprecedented colloidal patterns and assembly dynamics. Such a reconfiguration of the particle manipulation mechanism can be explained by the controllable competition or synergism between the coexisting acoustic and electric fields. Remarkably, in the EHD regime, a virtual zero-boundary electric quadrupole is created, and a novel colloidal diamond-shaped assembly is observed in this piezoelectric-quadrupole trap, which was rarely reported in acoustic or electric microfluidics. The presented in situ control of polarization revolutionizes our understanding of SAW and acoustofluidics, expands its potential by assuming the advantages of AHD and EHD on demand, and inspires new strategies in micro- and nanoscale manufacturing and manipulation, with applications beyond fundamental scientific interest.

关键词： Microfluidics

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Assessing the Impact of Sociotechnical Harms in AI-Based Medical Image Analysis 2nd

Assessing the Impact of Sociotechnical Harms in AI-Based ...

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2nd International Workshop on Fairness of AI in Medical Imaging, FAIMI 2024, and 3rd International Workshop on Ethical and Philosophical Issues in Medical Imaging, EPIMI 2024, Held in Conjunction with the International Conference on Medical Image Computing and Computer Assisted Interventions, MICCAI 2024

作者： Stanley, Emma A. M. Souza, Raissa Winder, Anthony J. Wilms, Matthias Pike, G. Bruce Dagasso, Gabrielle Nielsen, Christopher MacEachern, Sarah J. Forkert, Nils D. Biomedical Engineering Graduate Program University of Calgary Calgary Canada Department of Radiology University of Calgary Calgary Canada Hotchkiss Brain Institute University of Calgary Calgary Canada Alberta Children’s Hospital Research Institute University of Calgary Calgary Canada Department of Pediatrics University of Calgary Calgary Canada

ISBN: (纸本)9783031727863

Clinical decision-making and radiology will inevitably be transformed by artificial intelligence (AI) in the coming years. The rapid adoption of AI in this domain, and in other everyday applications, has brought an increased awareness of the potential impacts and negative consequences that may occur throughout the sociotechnical systems that these technologies are implemented in. In this paper, we review and apply a previously published taxonomy of the sociotechnical harms of AI to investigate how these harms could manifest during the development and clinical implementation of AI-based medical image analysis. Through an illustrative case study example on computer-aided diagnosis using brain magnetic resonance imaging, we demonstrate how performing impact assessments of sociotechnical harms can assist in operationalizing the medical ethics principle of non-maleficence, thereby guiding the ethical development and implementation of AI technologies in healthcare. © The Author(s), under exclusive license to Springer Nature Switzerland AG 2025.

关键词： Medical imaging

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Hyperspectral Image Analysis Using Maximum Abundance Classification

Hyperspectral Image Analysis Using Maximum Abundance Classif...

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International Conference on Intelligent Control and Information Processing (ICICIP)

作者： Nian Zhang Sougre-Nonma Stephanie Rouamba Wagdy H. Mahmoud Lara A. Thompson Department of Electrical and Computer Engineering University of the District of Columbia Washington D.C. USA Department of Mechanical Engineering Biomedical Engineering Program University of the District of Columbia Washington D.C. USA

ISBN: (数字)9798331516147

ISBN: (纸本)9798331516154

This paper presents a satellite hyperspectral image processing method that utilizes a maximum abundance classifier to categorize different regions of hyperspectral images into ground truth classes. First, the class names for each endmember and their corresponding columns in the signature matrix are identified, followed by the visualization of their spectral profiles. Abundance maps for the endmembers are then generated using the fully constrained least squares (FCLS) method. Afterward, the maximum abundance classifier is applied, and the resulting classified image is displayed with color-coded pixels. The abundance maps illustrate the spatial distribution of endmembers across the hyperspectral image, where the abundance values of each pixel represent the proportion of each endmember present. By determining the highest abundance value for each pixel and assigning it to the corresponding endmember class, the pixels within the hyperspectral images are classified. Experimental results demonstrate that the proposed MAC method effectively handles mixed pixels. In addition, it can effectively deal with the mixed pixel problem in hyperspectral images because it identifies components by calculating the abundance values for each pixel rather than relying solely on single spectral features.

关键词： Visualization Image segmentation Satellites Graphical models Information processing Hyperspectral imaging Intelligent control Image classification Distribution functions

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Intelligent reflux and suction detection system for ventricular assist devices: in silico study

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Research on biomedical engineering 2025年第1期41卷 1-23页

作者： Santos, Bruno J. Cestari, Idágene A. Biomedical Engineering Graduate Program Polythecnic School of Engineering University of Sao Paulo Av. Prof. Luciano Gualberto Travessa 3 nº380 Sao Paulo05508-010 Brazil Department of Information Technology of Universidade São Judas Tadeu Rua Taquari nº546 Sao Paulo03166-000 Brazil Faculdade de Medicina Bioengineering Division - Heart Institute Hospital das Clinicas HCFMUSP Universidade de Sao Paulo LIM Bioengenharia Sao Paulo Sao Paulo Brazil

Introduction: Left ventricular assist devices (LVADs) serve both as a bridge to transplantation and as destination therapy for the treatment of congestive heart failure (CHF). However, the inability of the existing control strategies to automatically adapt LVAD flow according to hemodynamic changes can significantly impact patient survival and quality of life. Physiological control strategies for LVAD show promising results, with reflux and suction detection (RSD) increasing device safety. Methods: This study presents in silico results of an RSD system based on measurements of inlet and outlet pressures in continuous-flow LVAD. Two strategies were used to investigate control feasibility, safety, and adjustments to nonlinear variations and comprehensively assess the system’s state considering a structured algorithm (SA) and ensembles of AI models (eAIm): K-nearest neighbors (KNN), support vector machine (SVM), and artificial neural network (ANN). Results: The SA submodule achieved an accuracy of 99.66% in suction detection but showed limitations in reflux events, with 80.04% accuracy and an F1-Score of 70.4%. The KNN and SVM models demonstrated performance exceeding 96% for both events, exhibiting more excellent stability than the SA submodule. The ANN excelled with low variability and an RMSE of 0.07 in R1, though its suction accuracy (96.7%) was slightly lower than for reflux (99.48%). The KNN was the most effective model, achieving 99.66% accuracy in suction and 98.44% in reflux. The SVM also produced competitive results but with variability across evaluations. The eAIm model showed satisfactory precision (97.78% for suction and 97.14% for reflux), with variations depending on the scenario. The eAIm is recommended for optimization in precision-critical situations. Discussion: These strategies are designed to fulfill the proposal’s feasibility, flexibility, and safety requirements. They address the challenges of achieving consistent reproduction using an SA and the

关键词： Left ventricular assist devices

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Retinal oxygen extraction during systemic hypoxia assessed with laser-speckle flowgraphy and retinal oximetry

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ACTA OPHTHALMOLOGICA 2025年第S284期103卷

作者： Pai, Viktoria Lindner, Theresa Janku, Patrick Schmetterer, Leopold Schmidl, Doreen Garhofer, Gerhard Department of Clinical Pharmacology Medical University of Vienna Vienna Austria Singapore National Eye Centre Singapore Eye Research Institute Singapore Singapore Duke-NUS Medical School Ophthalmology and Visual Sciences Academic Clinical Program Singapore Singapore SERI-NTU Advanced Ocular Engineering (STANCE) Nanyang Technological University Singapore Singapore Center for Medical Physics and Biomedical Engineering Medical University of Vienna Vienna Austria Institute of Molecular and Clinical Ophthalmology Basel Switzerland School of Chemistry Chemical Engineering and Biotechnology Nanyang Technological University Singapore Singapore

Aims/Purpose: Retinal oxygen extraction has been found to be altered in several ocular and systemic diseases, such as diabetic retinopathy, glaucoma or neurodegenerative diseases. However, measurement techniques for the assessment of retinal oxygen extraction are limited because most of them are based on time-consuming and custom-built devices. In the present pilot study, we assessed retinal oxygen extraction during systemic hypoxia with a combination of two commercially available devices: Laser-Speckle Flowgraphy (LSFG) and retinal oximetry. Methods: Retinal blood flow and retinal oxygen saturation were measured in each vessel around the optic nerve head at the same position. Arterial and venous oxygen content was estimated from the measured retinal oxygen saturation parameters and the difference between these two parameters was calculated (cO 2 , DIFF ). By using mean vessel flow rate (MV) for retinal blood flow and cO 2 , DIFF , retinal oxygen extraction was determined. For each healthy study participant, baseline measurement and measurement during inhalation of 88% nitrogen in 12% oxygen were performed. Results: Ten (10) healthy subjects (4 females, 6 males) with a mean age of 27 ± 3 years were included in this study. During systemic hypoxia, a significant decrease in arterial oxygen content occurred ( p = 0.046) while venous oxygen content did not change, leading to a decrease in cO 2 , DIFF . As expected, this was accompanied by a significant increase in MV ( p = 0.014). No change in retinal oxygen extraction was observed (46 ± 8 a.u. at baseline vs. 46 ± 5 a.u. during hypoxia, p = 0.997). Conclusions: Breathing of 88% nitrogen in 12% oxygen caused a significant decrease in retinal arterial oxygen content. This was compensated by an increase in retinal blood flow in order to keep retinal oxygen extraction constant. This was physiologically expected, and these findings are consistent with the literature. Therefore, the presented approach for measurement of reti

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Mammographic Breast Positioning Assessment via Deep Learning 1st

Mammographic Breast Positioning Assessment via Deep Learnin...

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1st Deep Breast Workshop on AI and Imaging for Diagnostic and Treatment Challenges in Breast Care, Deep-Breath 2024

作者： Tanyel, Toygar Denizoglu, Nurper Seker, Mustafa Ege Alis, Deniz Cerekci, Esma Karaarslan, Ercan Aribal, Erkin Oksuz, Ilkay Biomedical Engineering Graduate Program Istanbul Technical University Istanbul Turkey Department of Radiology Acibadem Healthcare Group Istanbul Turkey Department of Radiology University of Wisconsin-Madison MadisonWI United States Department of Radiology School of Medicine Acibadem University Istanbul Turkey Sisli Hamidiye Etfal Training and Research Hospital Istanbul Turkey Department of Computer Engineering Istanbul Technical University Istanbul Turkey

ISBN: (纸本)9783031777882

Breast cancer remains a leading cause of cancer-related deaths among women worldwide, with mammography screening as the most effective method for the early detection. Ensuring proper positioning in mammography is critical, as poor positioning can lead to diagnostic errors, increased patient stress, and higher costs due to recalls. Despite advancements in deep learning (DL) for breast cancer diagnostics, limited focus has been given to evaluating mammography positioning. This paper introduces a novel DL methodology to quantitatively assess mammogram positioning quality, specifically in mediolateral oblique (MLO) views using attention and coordinate convolution modules. Our method identifies key anatomical landmarks, such as the nipple and pectoralis muscle, and automatically draws a posterior nipple line (PNL), offering robust and inherently explainable alternative to well-known classification and regression-based approaches. We compare the performance of proposed methodology with various regression and classification-based models. The CoordAtt UNet model achieved the highest accuracy of 88.63% ± 2.84 and specificity of 90.25% ± 4.04, along with a noteworthy sensitivity of 86.04% ± 3.41. In landmark detection, the same model also recorded the lowest mean errors in key anatomical points and the smallest angular error of 2.42∘. Our results indicate that models incorporating attention mechanisms and CoordConv module increase the accuracy in classifying breast positioning quality and detecting anatomical landmarks. Furthermore, we make the labels and source codes available to the community to initiate an open research area for mammography, accessible at https://***/tanyelai/deep-breast-positioning. © The Author(s), under exclusive license to Springer Nature Switzerland AG 2025.

关键词： Diseases

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