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Nature Biotechnology Aug 2020Chimeric antigen receptor (CAR) T cell therapy has shown promise in hematologic malignancies, but its application to solid tumors has been challenging. Given the unique...
Chimeric antigen receptor (CAR) T cell therapy has shown promise in hematologic malignancies, but its application to solid tumors has been challenging. Given the unique effector functions of macrophages and their capacity to penetrate tumors, we genetically engineered human macrophages with CARs to direct their phagocytic activity against tumors. We found that a chimeric adenoviral vector overcame the inherent resistance of primary human macrophages to genetic manipulation and imparted a sustained pro-inflammatory (M1) phenotype. CAR macrophages (CAR-Ms) demonstrated antigen-specific phagocytosis and tumor clearance in vitro. In two solid tumor xenograft mouse models, a single infusion of human CAR-Ms decreased tumor burden and prolonged overall survival. Characterization of CAR-M activity showed that CAR-Ms expressed pro-inflammatory cytokines and chemokines, converted bystander M2 macrophages to M1, upregulated antigen presentation machinery, recruited and presented antigen to T cells and resisted the effects of immunosuppressive cytokines. In humanized mouse models, CAR-Ms were further shown to induce a pro-inflammatory tumor microenvironment and boost anti-tumor T cell activity.
Topics: Animals; Cell Line, Tumor; Cell Survival; Humans; Immunotherapy; Immunotherapy, Adoptive; Lung Neoplasms; Macrophages; Mice; Microscopy, Video; Neoplasms; Neoplasms, Experimental
PubMed: 32361713
DOI: 10.1038/s41587-020-0462-y -
Progress in Biophysics and Molecular... Jan 2022Compressed sensing (CS) is a signal processing approach that solves ill-posed inverse problems, from under-sampled data with respect to the Nyquist criterium. CS... (Review)
Review
Compressed sensing (CS) is a signal processing approach that solves ill-posed inverse problems, from under-sampled data with respect to the Nyquist criterium. CS exploits sparsity constraints based on the knowledge of prior information, relative to the structure of the object in the spatial or other domains. It is commonly used in image and video compression as well as in scientific and medical applications, including computed tomography and magnetic resonance imaging. In the field of fluorescence microscopy, it has been demonstrated to be valuable for fast and high-resolution imaging, from single-molecule localization, super-resolution to light-sheet microscopy. Furthermore, CS has found remarkable applications in the field of mesoscopic imaging, facilitating the study of small animals' organs and entire organisms. This review article illustrates the working principles of CS, its implementations in optical imaging and discusses several relevant uses of CS in the field of fluorescence imaging from super-resolution microscopy to mesoscopy.
Topics: Algorithms; Animals; Magnetic Resonance Imaging; Microscopy, Fluorescence; Optical Imaging; Signal Processing, Computer-Assisted
PubMed: 34153330
DOI: 10.1016/j.pbiomolbio.2021.06.004 -
Journal of Visualized Experiments : JoVE Mar 2023Aishwarya, R. et al. Visualizing subcellular localization of a protein in the heart using quantum dots-mediated immuno-labeling followed by transmission electron...
Aishwarya, R. et al. Visualizing subcellular localization of a protein in the heart using quantum dots-mediated immuno-labeling followed by transmission electron microscopy. Journal of Visualized Experiments. (187), e64085 (2022). Liu, B. et al. Visualizing mitophagy with fluorescent dyes for mitochondria and lysosome. Journal of Visualized Experiments. (189), e64647 (2022). Liu, C. Y., Zhu, J., Xie, Z. Visualizing yeast organelles with fluorescent protein markers. Journal of Visualized Experiments. (182), e63846 (2022). Zhao, X., Guo, S. Antibody uptake assay for tracking Notch/Delta endocytosis during the asymmetric division of zebrafish radial glia progenitors. Journal of Visualized Experiments. 65030 (2022). Park, S., Zhuang, Y., Shi, X. Label-retention expansion microscopy (LR-ExM) enables super-resolution imaging and high-efficiency labeling. Journal of Visualized Experiments. (188), e63793 (2022).
Topics: Animals; Zebrafish; Lysosomes; Mitochondria; Fluorescent Dyes; Microscopy, Electron, Transmission
PubMed: 37602874
DOI: 10.3791/64966 -
Indian Journal of Ophthalmology Jun 2023UBM is a high-resolution ultrasound technique which allows non-invasive, in vivo imaging of the anterior segment and iridocorneal angle.
BACKGROUND
UBM is a high-resolution ultrasound technique which allows non-invasive, in vivo imaging of the anterior segment and iridocorneal angle.
PURPOSE
This video is compilation of short video clips and images which gives description on identification of angle closure due to pupillary block, peripheral anterior synechiae, iris bombe, plateau iris, supra ciliary effusion, and malignant glaucoma. It also shows video demonstrating partially and fully patent iridotomy and features of trabeculectomy bleb. Synopsis: This video summarizes importance of UBM application in angle closure glaucoma to understand its pathophysiology by showing the relationship between the peripheral iris, trabecular meshwork and ciliary processes.
HIGHLIGHTS
UBM provides two-dimensional, grayscale images of the angle structures and allows identification of non-pupillary block mechanism in angle closure glaucoma, which can be recorded for qualitative and quantitative analyses.
VIDEO LINK
https://youtu.be/prsmGnR8jYc.
Topics: Humans; Ciliary Body; Eye Abnormalities; Glaucoma; Glaucoma, Angle-Closure; Iris; Iris Diseases; Microscopy, Acoustic; Pupil Disorders
PubMed: 37322722
DOI: 10.4103/IJO.IJO_153_23 -
Biomedical Optics Express Jul 2022Optical-resolution photoacoustic microscopy (OR-PAM) is widely utilized in biomedical applications because of its ability to noninvasively image biological tissues...
Optical-resolution photoacoustic microscopy (OR-PAM) is widely utilized in biomedical applications because of its ability to noninvasively image biological tissues while providing high-resolution morphological and functional information. However, one drawback of conventional OR-PAM is its imaging speed, which is restricted by the scanning technique employed. To achieve a higher imaging frame rate, we present video-rate high-resolution single-pixel nonscanning photoacoustic microscopy (SPN-PAM), which utilizes Fourier orthogonal basis structured planar illumination to overcome the above-mentioned limitations. A 473 × 473 µm imaging field of view (FOV) with 3.73 µm lateral resolution and video-rate imaging of 30 Hz were achieved. In addition, in both cell and mouse vascular hemodynamic imaging experiments, high-quality images were obtained at ultralow sampling rates. Thus, the proposed high-resolution SPN-PAM with video-rate imaging speed provides new insights into high-speed PA imaging and could be a powerful tool for rapid biological imaging.
PubMed: 35991922
DOI: 10.1364/BOE.459363 -
Biophysical Reports Jun 2024We present a method for tracking densely clustered, high-velocity, indistinguishable objects being spawned at a high rate and moving in a directed force field using only...
We present a method for tracking densely clustered, high-velocity, indistinguishable objects being spawned at a high rate and moving in a directed force field using only object centroids as inputs and no other image information. The algorithm places minimal restrictions on the velocities or accelerations of the objects being tracked and uses a methodology based on a scoring function and a backtracking refinement process. This combination leads to successful tracking of hundreds of particles in challenging environments even when the displacement of the individual objects at successive times approaches the separation between neighboring objects in any one frame. We note that these cases can be particularly difficult to handle by existing methods. The performance of the algorithm is methodically examined by comparison to simulated trajectories, which vary the temporal and spatial densities, velocities, and accelerations of the objects in motion, as well as the signal/noise ratio. Also, we demonstrate its capability by analyzing data from experiments with superparamagnetic microspheres moving in an inhomogeneous magnetic field in aqueous buffer at room temperature. Our method should be widely applicable since trajectory determination problems are ubiquitous in video microscopy applications in biology, materials science, physics, and engineering.
PubMed: 38505834
DOI: 10.1016/j.bpr.2024.100148 -
Microcirculation (New York, N.Y. : 1994) Oct 2022The aim of this study was to describe possible remodeling (i.e., dilatation and elongation) of papillary capillaries induced by increased oxygen demand for the repair...
OBJECTIVE
The aim of this study was to describe possible remodeling (i.e., dilatation and elongation) of papillary capillaries induced by increased oxygen demand for the repair process following a skin wound.
METHODS
Computer-assisted video microscopy was used to examine 10 healthy volunteers before (baseline) and after (≈1 h and ≈24 h) an incision (5 mm long and 1 mm deep) on the forearm, 0-1 mm and 30 mm (control site) from the incision. We defined categories from 0 (low) to 3 (high) to grade dilatation and elongation of the nutritive papillary capillaries, as well as the visibility of the superficial vascular plexus. Approximately 10 000 capillaries from 200 films were scored.
RESULTS
The nutritive papillary capillaries were dilated and elongated (p < 0.01) after ≈24 h; that is, elongation (score 1.9 ± 0.9) vs baseline (score 0.9 ± 0.6), p < 0.01 and dilatation (score 2.2 ± 0.7) vs baseline (score 0.3 ± 0.3), p < 0.01. Superficial plexus visibility increased (p < 0.01) after ≈1 h (score 2.0 ± 0.7) and ≈24 h (score 2.7 ± 0.3) vs baseline (score 0.8 ± 0.4).
CONCLUSION
The superficial vascular skin plexus showed enhanced visibility already ≈1 h after the skin trauma. Morphological remodeling in the nutritive papillary capillaries-dilatation and elongation after ≈24 h-facilitate increased O supply.
Topics: Humans; Microcirculation; Capillaries; Skin; Microscopy, Video; Forearm
PubMed: 35231135
DOI: 10.1111/micc.12755 -
International Journal of Molecular... Jan 2023The radial spoke head protein 4 homolog A () gene is one of more than 50 genes that cause Primary ciliary dyskinesia (PCD), a rare genetic ciliopathy. Genetic mutations... (Review)
Review
The radial spoke head protein 4 homolog A () gene is one of more than 50 genes that cause Primary ciliary dyskinesia (PCD), a rare genetic ciliopathy. Genetic mutations in the gene alter an important protein structure involved in ciliary pathogenesis. Radial spoke proteins, such as RSPH4A, have been conserved across multiple species. In humans, ciliary function deficiency caused by pathogenic variants results in a clinical phenotype characterized by recurrent oto-sino-pulmonary infections. More than 30 pathogenic genetic variants have been associated with PCD. In Puerto Rican Hispanics, a founder mutation ( (c.921+3_921+6delAAGT (intronic)) has been described. The spectrum of the PCD phenotype does not include laterality defects, which results in a challenging diagnosis. PCD diagnostic tools can combine transmission electron microscopy (TEM), nasal nitric oxide (nNO), High-Speed Video microscopy Analysis (HSVA), and immunofluorescence. The purpose of this review article is to provide a comprehensive overview of current knowledge about the gene in PCD, ranging from basic science to human clinical phenotype.
Topics: Humans; Kartagener Syndrome; Cilia; Proteins; Mutation; Axoneme; Cytoskeletal Proteins
PubMed: 36768259
DOI: 10.3390/ijms24031936 -
Indian Journal of Ophthalmology May 2023Ultrasound biomicroscopy (UBM) is a high-resolution ultrasound technique, which allows noninvasive, in vivo imaging of the ocular anterior segment structures.
BACKGROUND
Ultrasound biomicroscopy (UBM) is a high-resolution ultrasound technique, which allows noninvasive, in vivo imaging of the ocular anterior segment structures.
PURPOSE
This video gives a description of the identification of the iridocorneal angle structures in the cross-sectional view in a radial scan through a typical ciliary process and a guide toward measuring the angle parameters.
SYNOPSIS
UBM provides two-dimensional, grayscale images of the iridocorneal angle. The real-time image is displayed on a video monitor and can be recorded for qualitative and quantitative analysis. The angle parameters can be measured by in-built calipers in the machine software and manipulated by the examiner. Highlights: This video demonstrates UBM caliper positions as displayed on the monitor and marked by the examiner for the measurement of various anterior segment parameters of the eye.
VIDEO LINK
https://youtu.be/WTXMupYoyww.
Topics: Humans; Anterior Chamber; Anterior Eye Segment; Ciliary Body; Cross-Sectional Studies; Microscopy, Acoustic
PubMed: 37203007
DOI: 10.4103/IJO.IJO_3369_22 -
BioTechniques Feb 2022The use of magnetic tweezers for single-molecule micromanipulation has evolved rapidly since its introduction approximately 30 years ago. Magnetic tweezers have... (Review)
Review
The use of magnetic tweezers for single-molecule micromanipulation has evolved rapidly since its introduction approximately 30 years ago. Magnetic tweezers have provided important insights into the dynamic activity of DNA-processing enzymes, as well as detailed, high-resolution information on the mechanical properties of DNA. These successes have been enabled by major advancements in the hardware and software components of these devices. These developments now allow for a much richer mechanistic understanding of the functions and mechanisms of DNA-binding enzymes. In this review, the authors briefly discuss the fundamental principles of magnetic tweezers and describe the advancements that have made it a superlative tool for investigating, at the single-molecule level, DNA and its interactions with DNA-binding proteins.
Topics: DNA; Magnetic Phenomena; Magnetics; Micromanipulation; Nanotechnology; Optical Tweezers
PubMed: 35037472
DOI: 10.2144/btn-2021-0104