Critical Roles of microRNA-141-3p and CHD8 in Hypoxia/Reoxygenation-Induced Cardiomyocyte Apoptosis

Background: Cardiovascular diseases are currently the leading cause of death in humans. The high mortality of cardiac diseases is associated with myocardial ischemia and reperfusion (I/R). Recent studies have reported that microRNAs (miRNAs) play important roles in cell apoptosis. However, it is not known yet whether miR-141-3p contributes to the regulation of cardiomyocyte apoptosis. It has been well established that in vitro hypoxia/reoxygenation (H/R) model can follow in vivo myocardial I/R injury. This study aimed to investigate the effects of miR-141-3p and CHD8 on cardiomyocyte apoptosis following H/R. Results: We found that H/R remarkably reduces the expression of miR-141-3p but enhances CHD8 expression both in mRNA and protein in H9c2 cardiomyocytes. We also found either overexpression of miR-141-3p by transfection of miR-141-3p mimics or inhibition of CHD8 by transfection of small interfering RNA (siRNA) significantly decrease cardiomyocyte apoptosis induced by H/R. Moreover, miR-141-3p interacts with CHD8. Furthermore, miR-141-3p and CHD8 reduce the expression of p21. Conclusion: MiR-141-3p and CHD8 play critical roles in cardiomyocyte apoptosis induced by H/R. These studies suggest that miR-141-3p and CHD8 mediated cardiomyocyte apoptosis may offer a novel therapeutic strategy against myocardial I/R injury-induced cardiovascular diseases

GPX8 regulates pan-apoptosis in gliomas to promote microglial migration and mediate immunotherapy responses

IntroductionGliomas have emerged as the predominant brain tumor type in recent decades, yet the exploration of non-apoptotic cell death regulated by the pan-optosome complex, known as pan-apoptosis, remains largely unexplored in this context. This study aims to illuminate the molecular properties of pan-apoptosis-related genes in glioma patients, classifying them and developing a signature using machine learning techniques.MethodsThe prognostic significance, mutation features, immunological characteristics, and pharmaceutical prediction performance of this signature were comprehensively investigated. Furthermore, GPX8, a gene of interest, was extensively examined for its prognostic value, immunological characteristics, medication prediction performance, and immunotherapy prediction potential. ResultsExperimental techniques such as CCK-8, Transwell, and EdU investigations revealed that GPX8 acts as a tumor accelerator in gliomas. At the single-cell RNA sequencing level, GPX8 appeared to facilitate cell contact between tumor cells and macrophages, potentially enhancing microglial migration. ConclusionsThe incorporation of pan-apoptosis-related features shows promising potential for clinical applications in predicting tumor progression and advancing immunotherapeutic strategies. However, further in vitro and in vivo investigations are necessary to validate the tumorigenic and immunogenic processes associated with GPX8 in gliomas

Global Acoustic Speech Temporal Characteristics for Mandarin Speakers With Parkinson\u27s Disease During Syllable Repetition and Passage Reading

PURPOSE: Previous research has revealed considerable variation in speech rates among English speakers with Parkinson\u27s disease (PD) with slower, faster, or similar rates than controls. The purpose of this study was to characterize speech rates of Mandarin speakers with PD and the corresponding articulation and pause characteristics explaining the speech rates to enhance rate control therapies. METHOD: Eighteen Mandarin speakers with PD and 18 controls produced syllable repetitions and passage reading using their typical speech style. Speech rates, articulation rates, mean pause durations (≥ 10 ms), and the number of pauses with duration between 10 ms and 300 ms and greater than 300 ms were measured and compared between groups and tasks as well as across the initial, middle, and final periods of the passage. Two-way, mixed-model analyses of variance were employed for data analysis. RESULTS: Compared to controls, individuals with PD spoke with similar speech rates and faster articulation rates during passage reading, whereas during syllable repetitions, they produced slower speech rates and comparable articulation rates. The slower syllable repetitions produced by speakers with PD may be explained by longer pauses and more perceptual pauses, whereas faster articulation rates may explain the trend of faster speech rates during reading. Speech and articulation rates accelerated for both groups during passage reading. CONCLUSIONS: Speech rates of Mandarin speakers with PD were characterized by faster articulation, longer pauses, and more perceptual pauses for passage reading. A descriptive model of speech rate suggested that speakers with PD and dysarthria in this study would benefit from rate reduction therapy decreasing articulation rate. SUPPLEMENTAL MATERIAL: https://doi.org/10.23641/asha.23982282

Critical role of Tim-3 mediated autophagy in chronic stress induced immunosuppression

Abstract Background Psychological and physical stress can either enhance or suppress immune functions depending on a variety of factors such as duration and severity of stressful situation. Chronic stress exerts a significantly suppressive effect on immune functions. However, the mechanisms responsible for this phenomenon remain to be elucidated. Autophagy plays an essential role in modulating cellular homeostasis and immune responses. However, it is not known yet whether autophagy contributes to chronic stress-induced immunosuppression. T cell immunoglobulin and mucin domain 3 (Tim-3) has shown immune-suppressive effects and obviously positive regulation on cell apoptosis. Tim-3 combines with Tim-3 ligand galectin-9 to modulate apoptosis. However, its impact on autophagy and chronic stress-induced immunosuppression is not yet identified. Results We found remarkably higher autophagy level in the spleens of mice that were subjected to chronic restraint stress compared with the control group. We also found that inhibition of autophagy by the autophagy inhibitor 3-methyladenine (3-MA) significantly attenuated chronic stress-induced alterations of pro-inflammatory and anti-inflammatory cytokine levels. We further elucidated that 3-MA dramatically inhibited the reduction of lymphocyte numbers. Moreover, chronic stress dramatically enhanced the expression of Tim-3 and galectin-9. Inhibition of Tim-3 by small interfering RNA against Tim-3 significantly decreased the level of autophagy and immune suppression in isolated primary splenocytes from stressed mice. In addition, α-lactose, a blocker for the interaction of Tim-3 and galectin-9, also decreased the autophagy level and immune suppression. Conclusion Chronic stress induces autophagy, resulting with suppression of immune system. Tim-3 and galectin-9 play a crucial regulatory role in chronic stress-induced autophagy. These studies suggest that Tim-3 mediated autophagy may offer a novel therapeutic strategy against the deleterious effects of chronic stress on the immune system

Thermosensitive porphyrin-incorporated hydrogel with four-arm PEG-PCL copolymer (II): doxorubicin loaded hydrogel as a dual fluorescent drug delivery system for simultaneous imaging tracking in vivo

Visualization of a drug delivery system could reveal the pharmacokinetic properties, which is essential for the design of a novel drug delivery system. In vivo optical imaging offers an advanced tool to monitor the drug release process and the therapeutic effect by the combination of fluorescence imaging and bioluminescence imaging. Multispectral fluorescence imaging can separate the drug and the carrier without interference. Herein, a dual fluorescent anti-tumor drug delivery system was monitored with the doxorubicin-loaded hydrogel to further explore the application of the porphyrin-incorporated hydrogel with four-arm PEG-PCL copolymer as a drug carrier, based on the beneficial fluorescence and good biocompatibility of the porphyrin incorporated hydrogel. Using nude mice bearing luciferase expressed hepatic tumor as models, the whole process from the drug delivery to the tumor therapeutic effects were real time visualized simultaneously after administration at interval from 0 to 18 d. The imaging results suggest that the fluorescence signals of the drug and the carrier can be separated and unmixed from the drug-loaded hydrogel successfully, avoiding the interference of the fluorescence signals. The tumor growth or inhibition can be real time tracked and analyzed quantitatively by bioluminescence imaging. Noninvasive continuous tracking the in vivo drug delivery process simultaneously is a potential trend for the precise drug delivery and treatment

Dual actions of Netrin-1 on islet insulin secretion and immune modulation

Abstract Netrin-1 is typically known as a neural guidance cue, which has been implicated in pancreas development. Since regenerative, angiogenic and anti-inflammatory properties of Netrin-1 have been reported in multiple tissues, we have investigated the potential role of Netrin-1 in the endocrine islet and its implication in mice with high-fat diet (HFD)/streptozotocin (STZ)-induced diabetes. Effects of exogenous Netrin-1 on β-cell [Ca 2+ ] i , cyclic AMP (cAMP) and insulin production were assessed in vitro. The long-term impact of Netrin-1 treatment was then evaluated in HFD/STZ-induced diabetic mice by subcutaneous implantation of osmotic minipumps which release Netrin-1 in a sustained manner for 4 weeks. Immunostaining of pancreases of Netrin-1-treated and control animals were employed to examine islet morphology, vascularization and macrophage infiltration. Plasma insulin, glucagon and pro-inflammatory cytokine concentrations were quantified by ELISA. Expression of endogenous Netrin-1 was also assessed by PCR and immunohistochemistry. We observed a stimulatory effect of Netrin-1 on in vitro insulin secretion by promoting β-cell Ca 2+ influx and cAMP production. After 4-week continuous exposure, a hypoglycaemic property of Netrin-1 was demonstrated, which is probably attributable to improved β-cell function, shown as increased insulin content and preproinsulin mRNA expression. Enhanced islet vascularization, reduced islet macrophage infiltration and ameliorated systemic inflammation were detected from HFD/STZ-induced diabetic mice after Netrin-1 administration. We propose a dual action of Netrin-1 in islets during pathophysiological hyperglycaemia: by maintaining insulin secretion while attenuating inflammation

Bnip3 Mediates Doxorubicin-Induced Cardiomyocyte Pyroptosis via Caspase-3/GSDME

Aims: This study was aimed to investigate the role of GSDME-mediated pyroptosis in cardiac injury induced by Doxorubicin (DOX), and to evaluate the role of BH3-only protein Bcl-2/adenovirus E1B 19-kDa-interacting protein 3 (Bnip3) in regulation of DOX-induced pyroptosis. Main methods: HL-1 cardiomyocytes and C57BL/6J mice were treated by DOX to establish DOX-induced cardiotoxicity in vitro and in vivo models, respectively. Cell transfection was applied to regulate the expression of caspase-3, GSDME and Bnip3. Western blot was used for measuring expression of protein level. LDH-cytotoxicity assay was used to detect the LDH release. The Flow cytometry analysis was used to detect the cell death. Echocardiography was used to determine the cardiac function. HE staining was used for observing pathological feature of heart tissues. Key findings: Our results showed that GSDME-mediated pyroptosis was involved in DOX-induced cardiotoxicity in vivo. We showed that HL-1 cardiomyocytes exposed to DOX exhibited morphological features of pyroptosis in vitro. We also showed that DOX induced activation of caspase-3 and eventually triggered GSDME-dependent pyroptosis, which was reduced by the silence or inhibitor of caspase-3. We further showed that knockdown of GSDME inhibited DOX-induced cardiomyocyte pyroptosis in vitro. Finally, DOX increased the expression of Bnip3, whereas silencing of Bnip3 blunted cardiomyocyte pyroptosis induced by DOX, which was regulated through caspase-3 activation and GSDME cleavage. Significance: Our findings revealed a novel pathway that cardiomyocyte pyroptosis is regulated through Bnip3-caspase-3-GSDME pathway following DOX treatment, suggesting that Bnip3-dependent pyroptosis may offer a novel therapeutic strategy to reduce cardiotoxicity induced by DOX

ACCEPTED MANUSCRIPT Clinical Science Dual Actions of Netrin-1 on Islet Insulin Secretion and Immune Modulation

Abstract Aims: Netrin-1 is typically known as a neural guidance cue, which has been implicated in pancreas development. Since regenerative, angiogenic and anti-inflammatory properties of Netrin-1 have been reported in multiple tissues, we have investigated the potential role of Netrin-1 in the endocrine islet and its implication in mice with high-fat diet (HFD)/streptozotocin (STZ)-induced diabetes. Methods: Effects of exogenous Netrin-1 on -cell [Ca 2+ ] i , cAMP and insulin production were assessed in vitro. Long-term impact of Netrin-1 treatment was then evaluated in HFD/STZ-induced diabetic mice by subcutaneous implantation of osmotic minipumps which release Netrin-1 in a sustained manner for 4 weeks. Immunostaining of pancreases of Netrin-1-treated and control animals were employed to examine islet morphology, vascularisation and macrophage infiltration. Plasma insulin, glucagon and pro-inflammatory cytokine concentrations were quantified by ELISA. Expression of endogenous Netrin-1 was also assessed by PCR and immunohistochemistry. Results: We observed a stimulatory effect of Netrin-1 on in vitro insulin secretion by promoting -cell Ca 2+ influx and cAMP production. After 4-week continuous exposure, a hypoglycaemic property of Netrin-1 was demonstrated, which is likely attributable to improved -cell function, shown as increased insulin content and preproinsulin mRNA expression. Enhanced islet vascularisation, reduced islet macrophage infiltration and ameliorated systemic inflammation were detected from HFD/STZ-induced diabetic mice after Netrin-1 administration. Conclusions: We propose here a dual action of Netrin-1 in islets during pathophysiological hyperglycaemia: by maintaining insulin secretion while attenuating inflammation. Keywords: insulin secretion, islets, Netrin-1, diabetes, macrophage infiltration Summary statement Netrin-1 is known as a guidance molecule for neural and vascular growth. Its anti-inflammatory and regenerative properties were later discovered. We demonstrate a preventive role of Netrin-1 against experimental diabetes progression by stimulating insulin secretion while ameliorating islet inflammation. Clinical Perspectives Diabetes mellitus is a chronic disorder currently afflicting 415 million adults world-wide. Type 2 diabetes (T2D) is the most prevalent form of diabetes, accounting for approximately 90% of all cases of diabetes. The chronic hyperglycaemia associated with T2D can often lead to the development of devastating secondary complications causing reduced quality of life, increased morbidity, premature mortality and substantial health-care expenditure. Netrins, especially Netrin-1, have been identified as neurite attractants responsible for axon guidance during development. They are a family of proteins that are structurally similar to the extracellular matrix protein laminin. Recent studies reported additional roles of Netrin-1 in tissue regeneration following injuries in multiple organs. Given the pro-angiogenic and anti-inflammatory effects of Netrin-1, we investigated its potential role in diabetes since the disorder is exacerbated by islet deterioration and chronic inflammation. Considering the heterogeneous nature of diabetes, in order to mimic clinical manifestation of the disease, instead of using the inbred obese db/db leptin receptor-deficient obese mice, we used the outbred ICR mice with high-fat-diet (HFD)-induced insulin insensitivity and glucose intolerance followed by gradual islet -cell damage caused by repetitive administration of low dose streptozotocin (STZ, a specific -cell toxin; please see main text for references). Subsequent observations demonstrated that 4-week administration of Netrin-1 had a hypoglycaemic impact accompanied by improved -cell function, better islet vascularisation and reduced islet macrophage infiltration. In summary, we report a preventive role of Netrin-1 against diabetes progression via dual actions on islet insulin secretion and inflammation. Since diabetes and its co-morbidities are associated with over-active inflammation, Netrin-1 could be a promising therapeutic candidate that reduces hypoglycaemia while also ameliorates inflammation for diabetic individuals. Introduction Netrin-1 belongs to a family of laminin-related proteins that were originally identified as extracellular guidance cues for axon migration Indeed, Netrin-1 has been shown to promote developmental and therapeutic angiogenesis via the DCC receptors Additional roles of Netrin-1 have also been implicated beyond neuronal development and angiogenesis. Thus, Netrin-1 administration protected mice against acute kidney injury [6], pulmonary damage Type 2 diabetes (T2D) is primarily caused by -cell deterioration and insulin resistance. Dysfunction and loss of -cells eventually lead to inadequate insulin secretion to counteract impaired peripheral insulin sensitivity, resulting in sustained hyperglycaemia. As a result, current anti-diabetic agents consist largely of insulin secretagogues and insulin sensitizers that were designed to increase endogenous insulin production and insulin sensitivity. Recently, it has been established that inflammation also plays a significant part in the pathogenesis of T2D, which is a major risk factor of multiple diseases including cardiovascular disorders and cancer. Since diabetes and its co-morbidities are associated with over-active inflammation, an ideal treatment for diabetic individuals should not only address hyperglycaemia, but also aim to normalise pathologically elevated inflammatory tone. Given the regenerative and anti-inflammatory properties of Netrin-1, we have examined the therapeutic potential of Netrin-1 against high-fat diet (HFD)/streptozotocin (STZ)-induced diabetes

Precise and Long-Term Tracking of Adipose-Derived Stem Cells and Their Regenerative Capacity via Superb Bright and Stable Organic Nanodots

Monitoring and understanding long-term fate and regenerative therapy of administrated stem cells in vivo is of great importance. Herein we report organic nanodots with aggregation-induced emission characteristics (AIE dots) for long-term tracking of adipose-derived stem cells (ADSCs) and their regenerative capacity in living mice. The AIE dots possess high fluorescence (with a high quantum yield of 25 +/- 1%), excellent biological and photophysical stabilities, low in vivo toxicity, and superb retention in living ADSCs with negligible interference on their pluripotency and secretome. These AIE dots also exhibit superior in vitro cell tracking capability compared to the most popular commercial cell trackers, PKH26 and Qtracker 655. In vivo quantitative studies with bioluminescence and GFP labeling as the controls reveal that the AIE dots can precisely and quantitatively report the fate of ADSCs and their regenerative capacity for 42 days in an ischemic hind limb bearing mouse model