A5026097992- Phosphodiesterase function and regulation
- Receptor Mechanisms and Signaling
- Advanced Fluorescence Microscopy Techniques
- Optical Imaging and Spectroscopy Techniques
- Nitric Oxide and Endothelin Effects
- Photoacoustic and Ultrasonic Imaging
- Peptidase Inhibition and Analysis
- Medicinal plant effects and applications
- Advanced Biosensing Techniques and Applications
- Cell Image Analysis Techniques
- Inflammatory mediators and NSAID effects
- Ion channel regulation and function
- Coagulation, Bradykinin, Polyphosphates, and Angioedema
- Neuroscience and Neuropharmacology Research
- Synthesis and biological activity
- Analytical Methods in Pharmaceuticals
- Ion Transport and Channel Regulation
- Extracellular vesicles in disease
- Spectroscopy Techniques in Biomedical and Chemical Research
- 3D Printing in Biomedical Research
- Neuroinflammation and Neurodegeneration Mechanisms
- Angiogenesis and VEGF in Cancer
- Spectroscopy and Chemometric Analyses
- Chemical Synthesis and Analysis
- Connexins and lens biology
University of South Alabama
2016-2025
Abstract Standard fluorescence microscopy approaches rely on measurements at single excitation and emission bands to identify specific fluorophores the setting of thresholds quantify fluorophore intensity. This is often insufficient reliably resolve fluorescent labels in tissues due high autofluorescence. Here we describe use hyperspectral analysis techniques fluorescently labeled cells highly autofluorescent lung tissue. approach allowed accurate detection green protein (GFP) spectra, even...
In the last 20 years there have been remarkable advances in our ability to track movement and activities of proteins within cells. This is largely due improved chemical probes fluorescent proteins, technical microscopy. A remaining challenge real-time multiplexed imaging. Excitation scan-based hyperspectral imaging (HSI) approaches are well suited for However, excitation HSI has not widely adopted, part a lack protocols selection combinations labels determining range wavelengths dichroic...
Hyperspectral imaging and analysis approaches offer accurate detection quantification of fluorescently-labeled proteins cells in highly autofluorescent tissues. However, selecting optimum acquisition settings for hyperspectral is often a daunting task. In this study, we compared two systems-a widefield system with acoustic optical tunable filter (AOTF) charge coupled device (CCD) camera, confocal diffraction gratings photomultiplier tube (PMT) array. We measured the effects parameters on...
Colorectal cancer (CRC) is the second leading cause of cancer-related deaths worldwide. The gold standard for diagnosis tissue biopsy during colonoscopy and subsequent histopathology. Limitations current techniques include turnaround time required histopathology limited ability to detect flat lesions due inadequate contrast provided by traditional white light endoscopy (WLE). focus this work was assess detection accuracy differentiating CRC noncancerous tissues using excitation-scanning...
Herein we describe a pathogenic role for the Pseudomonas aeruginosa type three secretion system (T3SS) needle tip complex protein, PcrV, in causing lung endothelial injury. We first established model which P. wild strain PA103 caused pneumonia-induced sepsis and distal organ dysfunction. Interestingly, derivative lacking its two known secreted effectors, ExoU ExoT [denoted (ΔU/ΔT)], also modest injury whereas an isogenic T3SS assembly protein (ΔPcrV)] did not. (ΔU/ΔT) infection neutrophil...
Abstract Cyclic AMP is a ubiquitous second messenger that orchestrates variety of cellular functions over different timescales. The mechanisms underlying specificity within this signaling pathway are still not well understood. Several lines evidence suggest the existence spatial cAMP gradients cells, and compartmentalization underlies pathway. However, to date, no studies have visualized in three dimensions (3D: x , y z ).This part due limitations FRET‐based sensors, specifically low...
Acid-sensing ion channels (ASICs) are the major proton receptor in brain and a key mediator of acidosis-induced neuronal injuries disease. Most published data on ASIC function came from studies performed mice, relatively little is known about potential differences between human mouse ASICs (hASIC mASIC, respectively). This information critical for us to better interpret functional importance Here, we examined expression acutely resected cortical tissue. Compared with cortex, tissue showed...
Patients who recover from nosocomial pneumonia oftentimes exhibit long-lasting cognitive impairment comparable with what is observed in Alzheimer's disease patients. We previously hypothesized that the lung endothelium contributes to infection-related neurocognitive dysfunction, because bacteria-exposed endothelial cells release a form(s) of cytotoxic tau sufficient impair long-term potentiation hippocampus. However, full-length and isoform(s) have yet be resolved it remains unclear whether...
Abstract Previous studies have reported that phosphodiesterase 10A (PDE10) is overexpressed in colon epithelium during early stages of tumorigenesis and essential for cancer cell growth. Here we describe a novel non-COX inhibitory derivative the anti-inflammatory drug, sulindac, with selective PDE10 activity, ADT 061. 061 potently inhibited growth cells expressing high levels PDE10, but not normal colonocytes do express PDE10. The concentration range by which was identical to concentrations...
Hyperspectral imaging (HSI) technology has been applied in a range of fields for target detection and mixture analysis. While HSI was originally developed remote sensing applications, modern uses include agriculture, historical document authentication, medicine. also shown great utility fluorescence microscopy. However, traditional microscopy systems have suffered from limited signal strength due to the need filter or disperse emitted light across many spectral bands. We previously...
Physiological function is regulated through cellular communication that facilitated by multiple signaling molecules such as second messengers. Analysis of signal dynamics obtained from cell and tissue imaging difficult because intricate spatially temporally distinct signals. Signal analysis tools based on static region interest may under- or overestimate signals in relation to size location. Therefore, we developed an algorithm for biological detection dynamic regions interest, where...
Activation of the inflammasome-caspase-1 axis in lung endothelial cells is emerging as a novel arm innate immune response to pneumonia and sepsis caused by Pseudomonas aeruginosa. Increased levels circulating autacoids are hallmarks induce physiological responses via cAMP signaling targeted cells. However, it unknown whether affects other functions, such P. aeruginosa-induced caspase-1 activation. Herein, we describe effects on activation using single cell flow cytometry-based assay....
Cyclic AMP is a second messenger that involved in wide range of cellular and physiological activities. Several studies suggest cAMP signals are compartmentalized, compartmentalization contributes to signaling specificity within the pathway. The development Fӧrster resonance energy transfer (FRET) based biosensors has furthered ability measure visualize cells. However, these measurements often confined two spatial dimensions, which may result misinterpretation data. To date, there have been...
Cyclic AMP (cAMP) is a ubiquitous second messenger known to differentially regulate many cellular functions over wide range of timescales. Several lines evidence have suggested that the distribution cAMP within cells not uniform, and compartmentalization largely responsible for signaling specificity pathway. However, date, no studies experimentally measured three dimensional (3D) distributions cells. Here we use both 2D 3D hyperspectral microscopy visualize gradients in endothelial from...
Here, we report the generation of a Cre-recombinase (iCre) transgenic rat, where iCre is driven using vascular endothelial-cadherin (CDH5) promoter. The CDH5 promoter was cloned from rat pulmonary microvascular endothelial cells and demonstrated ~60% similarity to murine counterpart. 2,508 bp, it extended 79 bp beyond transcription start site, 22,923 upstream translation site. novel codon-optimized subcloned into Sleeping Beauty transposon vector for transpositional transgenesis in...
Second messenger signaling is required for cellular processes. We previously reported that extracellular vesicles (EVs) from stimulated cultured endothelial cells contain the biochemical second messenger, cAMP. In current study, we sought to determine whether cAMP-enriched EVs induce pathways in naïve recipient cells. Our results indicate increase cAMP content sufficient stimulate PKA activity. The implications of our work are represent a novel intercellular mechanism specifically cAMP, signaling.
Over the past 2 decades, hyperspectral imaging technologies have been adapted to address need for molecule-specific identification in biomedical field. Applications ranged from single-cell microscopy whole-animal vivo and basic research clinical systems. Enabling this growth has availability of faster, more effective filtering sensitive detectors. Hence, potential is high, many options are already commercially available. However, despite imaging, little work done aid users instruments...
Hyperspectral imaging (HSI) technologies have enabled a range of experimental techniques and studies in the fluorescence microscopy field. Unfortunately, drawback many HSI microscope platforms is increased acquisition time required to collect images across spectral bands, as well signal loss due need filter or disperse emitted into discrete bands. We previously demonstrated that an alternative approach scanning excitation spectrum can greatly improve system efficiency by decreasing light...
SignificanceHyperspectral imaging (HSI) technologies offer great potential in fluorescence microscopy for multiplexed imaging, autofluorescence removal, and analysis of autofluorescent molecules. However, there are also associated trade-offs when implementing HSI systems, such as decreased acquisition speed, resolution, or field-of-view due to the need acquire spectral information addition spatial information. The vast majority systems provide discrimination by filtering dispersing emission,...
Förster resonance energy transfer (FRET) is a valuable tool for measuring molecular distances and the effects of biological processes such as cyclic nucleotide messenger signaling protein localization. Most FRET techniques require two fluorescent proteins with overlapping excitation/emission spectral pairing to maximize detection sensitivity efficiency. microscopy often utilizes differing peak intensities selected fluorophores measured through different optical filter sets estimate index or...
A ubiquitous second messenger molecule, cAMP is responsible for orchestrating many different cellular functions through a variety of pathways. Fӧrster resonance energy transfer (FRET) probes have been used to visualize spatial gradients in pulmonary microvascular endothelial cells (PMVECs). However, FRET inherently low signal-to-noise ratios; multiple sources noise can obscure accurate visualization using hyperspectral imaging system. also measure 3D; however, it be difficult differentiate...
Studies of the cAMP signaling pathway have led to hypothesis that localized signals regulate distinct cellular responses. Much this work focused on measurement using sensors based upon Fӧrster resonance energy transfer (FRET). FRET-based probes are comprised a binding domain sandwiched between donor and acceptor fluorophores. Binding triggers conformational change which alters FRET efficiency. In order study signals, investigators targeted subcellular domains. This approach allows detection...
In the last 30 years data from a variety of studies have suggested that localized cAMP signals underlie signal specificity in cells and tissues. For example, pulmonary microvascular endothelial (PMVECs) produced by plasma membrane‐localized adenylyl cyclase is barrier protective, whereas both endogenous heterologously‐expressed AC disruptive. However, subcellular gradients been difficult to measure living cells. Most single cell approaches for measuring levels rely upon protein‐based probes....
Förster Resonance energy transfer (FRET) imaging has become a standard method for assessing protein‐protein interactions as well changes in the concentration of signaling molecules, such cAMP. However, many FRET probes or pairs have low signal‐to‐noise ratios and limited dynamic range. Our previous work suggested that hyperspectral analysis allows quantitative measurement efficiency 3D (x, y, t) within single cell. Here, we present data demonstrating approaches allow 4D z, assessment living...