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Biology subjects

Yang, H.-y.

Publications and source records attributed to Yang, H.-y..

7 recordsLinked to original sources

Glucose is a ligand-like trigger for secretion of postprandial GLP-1

Glucagon-like peptide 1 (GLP-1) is secreted from the intestinal L cells in a glucose-dependent manner. However, the mechanism underlying how glucose stimulates the secretion remains unclear. Here, we found that glucose at postprandial concentrations near the Km of GK for glucose ([~]10 mM), as well as nonmetabolizable glucose analogs, triggers GLP-1 secretion by intestinal L cells through directly binding to glucokinase (GK), without increasing intracellular ATP levels. We also show that in postprandially high glucose, glucose-bound GK interacts and inhibits KATP, as assessed by immunoprecipitation and FRET-FLIM analyses, as well as patch clamping. In addition, we have utilized GK mutants identified in diabetics, demonstrating that those mutants defective in binding to glucose impair GLP-1 secretion in however high glucose, whereas mutants constitutively bound to glucose with defects in catalysis, promote GLP-1 secretion even in low glucose. Furthermore, the KATP mutant unable to bind to GK blocks GLP-1 secretion in high glucose. We have also provided evidence showing that sodium-glucose cotransporter 1 (SGLT1) plays an important role in the basal secretion of GLP-1, but not in high glucose. We have thus revealed that glucose the molecule itself and GK act as a ligand and the receptor, respectively, to induce GLP-1 secretion in response to high glucose, providing new therapeutic avenues targeting metabolic diseases.

biochemistry↗

Scutellarin inhibits ferroptosis by promoting cellular antioxidant capacity through regulating the Nrf2 signaling

Ferroptosis is a lytic form of regulated cell death that is driven by iron-dependent lipid peroxidation, and has been implicated in various diseases including acute kidney injury (AKI). Scutellarin is a flavonoid isolated from Erigeron breviscapus (Vant.) Hand.-Mazz. and possesses various pharmacological activities including anti-inflammatory and antioxidative properties. Yet it is unclear whether scutellarin can inhibit ferroptosis and mitigate related diseases. In this study, we found that scutellarin was able to inhibit ferroptosis in both human HK-2 cells and mouse bone marrow-derived macrophages stimulated with RSL3 or erastin. Mitochondrial dysfunction and reactive oxygen species generation were counteracted by scutellarin treatment, suggesting involvement of its antioxidative activity. Furthermore, scutellarin increased the nuclear levels of Nrf2 and the expression of its target genes including HO-1 and GPX4. Scutellarin-mediated inhibition of ferroptosis and an increase of these proteins was abrogated by the co-treatment with brusatol, an Nrf2 inhibitor, indicating an essential role for Nrf2 in this process. In a mouse model of folic acid-induced AKI, scutellarin mitigated acute renal damage as revealed by histopathological analysis and serum blood urea nitrogen and creatinine assays. Folic acid-induced acute renal injury was associated with increased ferroptosis as revealed by elevated levels of 4-hydroxynonenal (4-HNE), a surrogate marker of ferroptosis, which were diminished by scutellarin cotreatment. Specifically, the elevated 4-HNE levels in macrophages (MAC-2 positive) and other renal cells were suppressed by scutellarin. Collectively, scutellarin can inhibit ferroptosis both in cultured cells and in a mouse model of AKI by regulating Nrf2 signaling.

pharmacology and toxicology↗

Design and Validation of a Wearable Imaging System for Automated Wound Monitoring in Porcine Model

Effective wound monitoring has the potential to guide treatment regiments and improve healing outcomes, yet current clinical assessment methods remain largely subjective and labor-intensive. To address this challenge, we present a high-resolution wearable imaging system designed for continuous wound monitoring. The system integrates a 64 MP camera with a plano-convex lens housed in an enclosure measuring 73 mm in diameter and 36.1 mm in height, and features a custom printed circuit board (PCB) for programmable LED illumination. The 3D-printed device enclosure is designed to accommodate a silicone bioelectronic device and can be securely attached using a commercially available ostomy skin barrier. In porcine model validation studies, the system successfully captured daily wound progression over periods up to 7 days. The captured images were wirelessly transmitted to a processing unit where DeepMapper, a machine learning algorithm, processed z-stacked images and performed multi-level feature extraction to predict wound healing stages and indicate potential complications such as infection. This imaging system enables automated analysis of wound progression and supports the development of smart wound care platforms for personalized treatment strategies. The integrated design approach demonstrates the feasibility of creating compact, high-resolution imaging systems suitable for clinical wound monitoring applications.

bioengineering↗

Wireless bioelectronic device for wound healing

Wireless bioelectronic actuators have been developed to deliver targeted treatments over multiple days while continuously monitoring delivery, thereby improving wound healing. Specifically, these devices can deliver charged biomolecules such as fluoxetine cations (Flx+) and electric field (EF) in freely moving pigs. Treatments can be controlled and monitored in real time via WiFi, with options for both user-specified delivery rates and durations, as well as automated closed-loop (CL) control. The devices are engineered to handle various failure scenarios that may arise in dynamic, real-world experiments--such as communication or power interruptions--ensuring that valuable experimental data is collected with minimal disruption. The ion pump features eight drug reservoirs and channels arranged around a 20 mm diameter-wound, with a central ground electrode (0 V). When voltages above 0 V are applied to the outer channels, currents flow from the reservoirs and channels into the wound, delivering Flx+ and/or EF depending on the reservoir solution. The device records applied voltages and currents locally to a microSD card at a high sampling rate, while simultaneously transmitting real-time measurements via a local WiFi network to a wound healing algorithm running on a nearby laptop. CL control of current/delivery rate is performed by an onboard microcontroller unit (MCU) and current-source microchips, based on instructions received from the wound healing algorithm. A graphical user interface (GUI) provides intuitive user control and real-time data visualization, with support for multiple devices. In vivo studies over seven days showed that Flx+-treated wounds had a 20% lower M1/M2 macrophage ratio and 41.67% greater re-epithelialization compared to controls (standard-of-care), demonstrating the actuators potential to enhance wound healing.

biochemistry↗

Programmable delivery of fluoxetine via wearable bioelectronics for wound healing in vivo

The ability to deliver drugs with precise dosages at specific time points can significantly improve disease treatment while reducing side effects. Drug encapsulation for gradual delivery has opened up the doors for superior treatment regimen. To expand on this ability, programming bioelectronic devices to deliver small molecules enables ad-hoc personalized therapeutic profiles that are more complex than simple gradual release. Here, we introduce a wearable bioelectronic bandage with an integrated electrophoretic ion pump that affords on-demand drug delivery with precise dose control. Delivery of fluoxetine to wounds in mice resulted in a 27.2% decrease in the macrophage ratio (M1/M2) and a 39.9% increase in re-epithelialization, indicating a shorter inflammatory phase and faster overall healing. Programmable drug delivery using wearable bioelectronics in wounds introduces a broadly applicable strategy for the long-term delivery of a prescribed treatment regimen with minimal external intervention.

bioengineering↗

Establishing cell motility patterns as predictors of macrophage subtypes and their relation to cell morphology

The motility of macrophages in response to microenvironment stimuli is a hallmark of innate immunity, where macrophages play pro-inflammatory or pro-reparatory roles depending on their activation status during wound healing. Cell size and shape have been informative in defining macrophage subtypes. Studies show pro and anti-inflammatory macrophages exhibit distinct migratory behaviors, in vitro, in 3D and in vivo but this link has not been rigorously studied. We apply both morphology and motility-based image processing approaches to analyze live cell images consisting of macrophage phenotypes. Macrophage subtypes are differentiated from primary murine bone marrow derived macrophages using a potent lipopolysaccharide (LPS) or cytokine interleukin-4 (IL-4). We show that morphology is tightly linked to motility, which leads to our hypothesis that motility analysis could be used alone or in conjunction with morphological features for improved prediction of macrophage subtypes. We train a support vector machine (SVM) classifier to predict macrophage subtypes based on morphology alone, motility alone, and both morphology and motility combined. We show that motility has comparable predictive capabilities as morphology. However, using both measures can enhance predictive capabilities. While motility and morphological features can be individually ambiguous identifiers, together they provide significantly improved prediction accuracies (75%) from a training dataset of 1000 cells tracked over time using only phase contrast time-lapse microscopy. Thus, the approach combining cell motility and cell morphology information can lead to methods that accurately assess functionally diverse macrophage phenotypes quickly and efficiently. This can support the development of cost efficient and high through-put methods for screening biochemicals targeting macrophage polarization. Author summaryPrevious work has shown that macrophage phenotypes can be distinguished by their morphological characteristics. We extend this work to show that distinct motility patterns are linked to macrophage morphology. Thus, motility patterns can be used to differentiate phenotypes. This can enable high-throughput classification of cell phenotypes without regard for the high-resolution images needed to quantify morphological characteristics. Furthermore, combining motility-based features with morphological information improves prediction of macrophage subtypes by a machine learning based classification model.

cell biology↗

Automatic wound detection and size estimation using deep learning algorithms

Evaluating and tracking wound size is a fundamental metric for the wound assessment process. Good location and size estimates can enable proper diagnosis and effective treatment. Traditionally, laboratory wound healing studies include a collection of images at uniform time intervals exhibiting the wounded area and the healing process in the test animal, often a mouse. These images are then manually observed to determine key metrics --such as wound size progress- relevant to the study. However, this task is a time-consuming and laborious process. In addition, defining the wound edge could be subjective and can vary from one individual to another even among experts. Furthermore, as our understanding of the healing process grows, so does our need to efficiently and accurately track these key factors for high throughput (e.g., over large-scale and long-term experiments). Thus, in this study, we develop a deep learning-based image analysis pipeline that aims to intake non-uniform wound images and extract relevant information such as the location of interest, wound only image crops, and wound periphery size over-time metrics. In particular, our work focuses on images of wounded laboratory mice that are used widely for translationally relevant wound studies and leverages a commonly used ring-shaped splint present in most images to predict wound size. We apply the method to a dataset that was never meant to be quantified and, thus, presents many visual challenges. Additionally, the data set was not meant for training deep learning models and so is relatively small in size with only 256 images. We compare results to that of expert measurements and demonstrate preservation of information relevant to predicting wound closure despite variability from machine-to-expert and even expert-to-expert. The proposed system resulted in high fidelity results on unseen data with minimal human intervention. Furthermore, the pipeline estimates acceptable wound sizes when less than 50% of the images are missing reference objects. Author summaryKnowledge of the wound size changes over-time allows us to observe important insights such as rate of closure, time to closure, and expansion events, which are key indicators for predicting healing status. To better perform wound measurements it is essential to utilize a technique that returns accurate and consistent results every time. Over the last years, collecting wound images is becoming easier and more popular as digital cameras and smartphones are more accessible. Commonly, scientists/clinicians trace the wound in these images manually to observe changes in the wound, which is normally a slow and labor-intensive process and also requires a trained eye. The clinical goal is to more efficiently and effectively treat wounds by employing easy to use and precise wound measurement techniques. Therefore, the objective should be devising automatic and precise wound measurement tools to be used for wound assessment. To this end, we leveraged a combination of various state-of-the-art computer vision and machine learning-based methods for developing a versatile and automatic wound assessment tool. We applied this tool to analyze the images of wound inflicted lab mice and showed that our developed tool automated the overall wound measurement process, therefore, resulting in high fidelity results without significant human intervention. Furthermore, we compared results to two expert measurements. We found variability in measurement even across experts further validating the need for a consistent approach. However, qualitative behavior, which is most important for predicting wound closure, is preserved.

bioinformatics↗