Telehealth of cardiac devices for CVD treatment.

This review covers currently available cardiac implantable electronic devices (CIEDs) as well as updated progress in real-time monitoring techniques for CIEDs. A variety of implantable and wearable devices that can diagnose and monitor patients with cardiovascular diseases are summarized, and various working mechanisms and principles of monitoring techniques for Telehealth and mHealth are discussed. In addition, future research directions are presented based on the rapidly evolving research landscape including Artificial Intelligence (AI).

Effects of volume-matched once-weekly and thrice-weekly high-intensity interval training (HIIT) on body adiposity in adults with central obesity: Study protocol for a randomized controlled trial.

Objective: This study aims to examine the comparative effects of 75 min of volume-matched once-weekly and thrice-weekly high-intensity interval training (HIIT) on body adiposity in adults with central obesity. Methods: This assessor-blinded, three-arm, randomized controlled trial will recruit 315 physically inactive adults with central obesity (aged ≥18 years, body mass index ≥23, waist circumference ≥90 cm for men and ≥80 cm for women). Participants will be randomly allocated to the once-weekly HIIT, thrice-weekly HIIT or usual care control group. Participants in the HIIT groups will receive weekly exercise training sessions for 16 weeks, prescribed either once or three times weekly. Each HIIT session will consist of a supervised program of four 4-min high-intensity intervals at 85%-95% peak heart rate (HRpeak) interspersed with 3-min active recovery intervals at 50%-70% HRpeak. Participants in the once-weekly HIIT group will perform the 25-min HIIT bout three times with a break between each 25-min HIIT bout. The usual care control group will receive bi-weekly health education classes. The outcome assessments will be conducted at baseline, 16 weeks (post-intervention) and 32 weeks (follow-up). The primary outcome will be total body adiposity assessed by dual-energy X-ray absorptiometry (DXA). The secondary outcome measures will include markers of cardiovascular and metabolic health (body composition, cardiorespiratory fitness, blood pressure, and blood lipids), mental health, cognitive performance, health-related quality of life, sleep quality, habitual physical activity, diet, medication, adverse events and adherence to the intervention. Impact of the project: The findings from this study are expected to consolidate the therapeutic efficacy of HIIT for the management of central obesity and inform the comparative compliance, feasibility and suitability of once-weekly and thrice-weekly HIIT as exercise strategies to manage obesity. In particular, the present study is expected to provide a novel perspective on the utility of low-frequency HIIT (i.e., once-weekly) as an effective and sustainable exercise strategy to tackle the obesity pandemic. The anticipated findings will hold substantial translational value by informing public health policies and enhancing exercise compliance in the physically inactive obese population. Trial registration: ClinicalTrials.gov (NCT04887454).

Direct Activation of Endothelial Cells by SARS-CoV-2 Nucleocapsid Protein Is Blocked by Simvastatin.

Emerging evidence suggests that endothelial activation plays a central role in the pathogenesis of acute respiratory distress syndrome (ARDS) and multiorgan failure in patients with coronavirus disease 2019 (COVID-19). However, the molecular mechanisms underlying endothelial activation in COVID-19 patients remain unclear. In this study, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral proteins that potently activate human endothelial cells were screened to elucidate the molecular mechanisms involved in endothelial activation. It was found that nucleocapsid protein (NP) of SARS-CoV-2 significantly activated human endothelial cells through Toll-like receptor 2 (TLR2)/NF-κB and mitogen-activated protein kinase (MAPK) signaling pathways. Moreover, by screening a natural microbial compound library containing 154 natural compounds, simvastatin was identified as a potent inhibitor of NP-induced endothelial activation. Remarkably, though the protein sequences of N proteins from coronaviruses are highly conserved, only NP from SARS-CoV-2 induced endothelial activation. The NPs from other coronaviruses such as SARS-CoV, Middle East respiratory syndrome coronavirus (MERS-CoV), HUB1-CoV, and influenza virus H1N1 did not activate endothelial cells. These findings are consistent with the results from clinical investigations showing broad endotheliitis and organ injury in severe COVID-19 patients. In conclusion, the study provides insights on SARS-CoV-2-induced vasculopathy and coagulopathy and suggests that simvastatin, an FDA-approved lipid-lowering drug, may help prevent the pathogenesis and improve the outcome of COVID-19 patients. IMPORTANCE Coronavirus disease 2019 (COVID-19), caused by the betacoronavirus SARS-CoV-2, is a worldwide challenge for health care systems. The leading cause of mortality in patients with COVID-19 is hypoxic respiratory failure from acute respiratory distress syndrome (ARDS). To date, pulmonary endothelial cells (ECs) have been largely overlooked as a therapeutic target in COVID-19, yet emerging evidence suggests that these cells contribute to the initiation and propagation of ARDS by altering vessel barrier integrity, promoting a procoagulative state, inducing vascular inflammation and mediating inflammatory cell infiltration. Therefore, a better mechanistic understanding of the vasculature is of utmost importance. In this study, we screened the SARS-CoV-2 viral proteins that potently activate human endothelial cells and found that nucleocapsid protein (NP) significantly activated human endothelial cells through TLR2/NF-κB and MAPK signaling pathways. Moreover, by screening a natural microbial compound library containing 154 natural compounds, simvastatin was identified as a potent inhibitor of NP-induced endothelial activation. Our results provide insights on SARS-CoV-2-induced vasculopathy and coagulopathy, and suggests that simvastatin, an FDA-approved lipid-lowering drug, may benefit to prevent the pathogenesis and improve the outcome of COVID-19 patients.

Enzyme Responsive Delivery of Anti-Retroviral Peptide via Smart Hydrogel.

In response to an urgent need for advanced formulations for the delivery of anti-retrovirals, a stimuli-sensitive hydrogel formulation that intravaginally delivers HIV-1 entry inhibitor upon being exposed to a specific protease was developed. The hydrogel formulation consists of PEG-azide and PEG-DBCO covalently linked to the entry inhibitor peptide, enfuvirtide, via substrate linker that is designed to undergo proteolysis by prostate specific antigen (PSA) present in seminal fluid and release innate enfuvirtide. Of the tested PSA substrate linkers (HSSKLQYY, GISSFYSSK, AYLMYY, and AYLMGRR), HSSKLQ was found to be an optimal candidate for PEG-based hydrogel with kcat/KM of 2.2 M-1 s-1. The PEG-based hydrogel displayed a pseudoplastic, thixotropic behavior with overall viscosity varying between 1516 and 2.2 Pa.s, within the biologically relevant shear rates of 0.01-100 s-1. It also exhibited viscoelastic properties appropriate for uniform spreading and being retained in vagina. PEG-based hydrogels were loaded with N3-HSSKLQ-enfuvirtide (HF42) that is customarily synthesized enfuvirtide prodrug with its N-terminus connected to HSSKLQ linker. The stimuli-sensitive PEG-based hydrogel formulations upon being exposed to PSA released 36.5 ± 4.8% of enfuvirtide over 24 h in human ejaculate mimic of vaginal simulant fluid and seminal simulant fluid mixed in 1:3 ratio, which is significantly greater than its IC50. The PEG-based hydrogel was non-cytotoxic to both vaginal epithelial cells (VK2/E6E7) and murine macrophages (RAW 264.7) and did not significantly induce the production of nitric oxide, an inflammatory mediator. The PEG-based hydrogel is found to have suitable physicochemical properties for an intravaginal formulation of the PSA substrate-linked anti-retrovirals and is safe towards vaginal epithelium. It is capable of delivering enfuvirtide with effective concentrations to prevent women from HIV-1 infection.

Effects of one-year once-weekly high-intensity interval training on body adiposity and liver fat in adults with central obesity: Study protocol for a randomized controlled trial.

Objective: This study aims to examine the effects of one-year, once-weekly high-intensity interval training (HIIT) on body adiposity and liver fat in adults with central obesity. Methods: One-hundred and twenty adults aged 18-60 years with central obesity (body mass index ≥25, waist circumference ≥90 cm for men and ≥80 cm for women). This is an assessor-blinded randomized controlled trial. Participants will be randomly assigned to the HIIT group or the usual care control group. Each HIIT session will consist of 4 × 4-min bouts at 85%-95% maximal heart rate, interspersed with 3-min bouts at 50%-70% maximal heart rate. The HIIT group will complete one session per week for 12 months, whereas the usual care control group will receive health education. The primary outcomes of this study are total body adiposity and intrahepatic triglyceride content. The secondary outcomes include abdominal visceral adipose tissue, subcutaneous adipose tissue, body mass index, waist circumference, hip circumference, cardiorespiratory fitness, lean body mass, bone mineral density, blood pressure, fasting blood glucose, insulin, triglycerides, glycated hemoglobin, cholesterol profile, liver function enzymes, medications, adherence to exercise, adverse events, quality of life, and mental health. Outcome measure will be conducted at baseline, 12 months (post-intervention), and 24 months (one-year follow-up). Impact of the project: This study will explore the benefits of long-term once-weekly HIIT with a follow-up period to assess its effectiveness, adherence, and sustainability. We expect this intervention will enhance the practical suitability of HIIT in inactive adults with central obesity, and provide insights on low-frequency HIIT as a novel exercise option for the management of patients with central obesity and liver fat. Trial registration: ClinicalTrials.gov (NCT03912272) registered on 11 April 2019.

Parametric Optimization of 3D Printed Hydrogel-Based Cardiovascular Stent.

PURPOSE: This study aimed to develop personalized biodegradable stent (BDS) for the treatment of coronary heart disease. Three-dimensional (3D) printing technique has offered easy and fast fabrication of BDS with enhanced reproducibility and efficacy. METHODS: A variety of BDS were printed with 3 types of hydrogel (~5 ml) resources (10%w/v sodium alginate (SA), 10%w/v cysteine-sodium alginate (SA-CYS), and 10%w/v cysteine-sodium alginate with 0.4%w/v PLA-nanofibers (SA-CYS-NF)) dispersed from an 22G print head nozzle attached to the BD-syringe. The printability of hydrogels into 3D structures was examined based on such variables as hydrogel's viscosity, printing distance, printing speed and the nozzle size. RESULTS: It was demonstrated that alginate composition (10%w/v) offered BDS with sufficient viscosity that defined the thickness and swelling ratio of the stent struts. The thickness of the strut was found to be 338.7 ± 29.3 µm, 262.5 ± 14.7 µm and 237.1 ± 14.7 µm for stents made of SA, SA-CYS and SA-CYS-NF, respectively. SA-CYS-NF stent displayed the highest swelling ratio of 38.8 ± 2.9% at the initial 30 min, whereas stents made of SA and SA-CYS had 23.1 ± 2.4% and 22.0 ± 2.4%, respectively. CONCLUSION: The printed stents had sufficient mechanical strength and were stable against pseudo-physiological wall shear stress. An addition of nanofibers to alginate hydrogel significantly enhanced the biodegradation rates of the stents. In vitro cell culture studies revealed that stents had no cytotoxic effects on human umbilical vein endothelial cells (HUVECs) and Raw 264.7 cells (i.e., Monocyte/macrophage-like cells), supporting that stents are biocompatible and can be explored for future clinical applications.

A Data-Mining Approach for the Quantitative Assessment of Physicochemical Properties of Molecular Compounds in the Skin Flux.

This paper aimed to provide an insight into the mechanism of transdermal penetration of drug molecules with respect to their physicochemical properties, such as solubility (S), the presence of enantiomer (ET) and logarithm of octanol-water partition coefficient (log P), molecular weight (MW), and melting point (MP). Propionic acid derivatives were evaluated for their flux through full-thickness skin excised from hairless mice upon being delivered from silicone-based pressure-sensitive adhesive (PSA) matrices in the presence or absence of various enhancers. The skin fluxes of model compounds were calculated based on the data obtained using the method engaged with the diffusion cell system. The statistical design of experiments (DoE) based on the factorial approach was used to find variables that have a significant impact on the outcomes. For the prediction of skin flux, a quantitative equation was derived using the data-mining approach on the relationship between skin permeation of model compounds (~125 mg/ml) and involved physicochemical variables. The most influential variables for the skin flux of propionic acid derivatives were the melting point (0.97) followed by the presence of enantiomer (0.95), molecular mass (0.93), log P values (0.86), and aqueous solubility (0.80). It was concluded that the skin flux of molecular compounds can be predicted based on the relationship between their physicochemical properties and the interaction with cofactors including additives and enhancers in the vehicles.

Gene expression profiling of lichenoid dermatitis immune-related adverse event from immune checkpoint inhibitors reveals increased CD14+ and CD16+ monocytes driving an innate immune response.

BACKGROUND: Cancer patients receiving antibodies abrogating immune checkpoint pathways may develop a diverse array of immune-related adverse events (irAEs), of which lichenoid dermatitis (LD) is the most common. The mechanism driving the emergence of these irAEs remain understudied, underscoring a critical need to determine the unique gene expression profiles and immune composition in LD-irAE. METHODS: LD-irAE (n = 3) and benign lichenoid keratosis (BLK) control (n = 3) were profiled with NanoString nCounter PanCancer Immune Profiling Panel interrogating the mRNA levels of 770 genes. Immunohistochemical (IHC) studies (n = 14 samples) for CD14, CD16, T-Bet, Gata-3, and FoxP3 were further evaluated using Aperio digital image analysis. RESULTS: The LD-irAE showed downregulation of 93 mRNA transcripts (P < 0.05) and upregulation of 74 mRNA transcripts (P < 0.04) including toll-like receptor (TLR) 2 and TLR4 (P < 0.05). CD14+ and CD16+ monocytes quantified by IHC (H-score) were higher in LD-irAE than in the BLK control (P < 0.05). The immune composition of LD-irAE exhibited higher numbers of T-Bet+ (Th1) cells compared with Gata-3+ (Th2) cells (P = 0.016) and lower numbers of FoxP3 (T regulatory) cells (P = 0.008). CONCLUSIONS: LD-irAE exhibited activation of CD14/TLR innate immune response with increased CD14+ and CD16+ monocytes compared with BLK control. CD14/TLR signaling may drive the development of LD-irAE.

Computational Analysis on Down-Regulated Images of Macrophage Scavenger Receptor.

BACKGROUND: Thiolated-graphene quantum dots (SH-GQDs) were developed and assessed for an efficient preventive means against atherosclerosis and potential toxicity through computational image analysis and animal model studies. EXPERIMENTS: Zebrafish (wild-type, wt) were used for evaluation of toxicity through the assessment of embryonic mortality, malformation and ROS generation. The amounts of SH-GQDs uptaken by mouse macrophage cells (Raw264.7) were analyzed using a flow cytometer. For the time-dependent cellular uptake study, Raw264.7 cells were treated with SH-GQDs (200 µg/ml) at specific time intervals (0.5, 1, 2, 5, 10 and 24 h). The efficacy of SH-GQDs on DiO-oxLDL efflux by Raw264.7 cells was evaluated (DiO, 3,3'-dioctadecyl-oxacarbocyanine) based on the percentage of positive cells containing DiO-oxLDL. TEER of human primary umbilical vein endothelial cells (hUVECs) were examined to assess the barrier function of the cell layers upon being treated with oxLDL. RESULTS: SH-GQDs significantly enhanced the efflux of oxLDL and down-regulated macrophage scavenger receptor (MSR) in Raw264.7. The ROS levels stimulated by oxidative stress were alleviated by SH-GQDs. oxLDL (10 µg/ml) significantly impaired the barrier function (TEER) of adherence junctions, which was recovered by SH-GQDs (10 µg/ml) (oxLDL: 67.2 ± 2.2 Ω-cm2 for 24 h; SH-GQDs: 114.6 ± 8.5 Ω-cm2 for 24 h). The mortality rate (46% for 1 mg/ml) of the zebra fish increased, as the concentrations and exposure duration of SH-GQDs increased. SH-GQDs exerted negligible side effects. CONCLUSION: SH-GQDs have target specificity to macrophage scavenger receptor (MSR) and efficiently recovered the ROS levels and TEER. SH-GQDs did not induce endothelial cell layer disruption nor affected zebrafish larvae survival.

Development of Thiolated-Graphene Quantum Dots for Regulation of ROS in macrophages.

PURPOSE: The current study was designed to develop thiolated-graphene quantum dots (SH-GQDs) as a theranostic nanocarrier and evaluate its potential for the optimal scavenging of reactive oxygen species (ROS) in macrophages. METHODS: SH-GQDs were prepared by hydrothermal pyrolysis of carbon source (citric acid) in the presence of reduced-glutathione (GSH). Raw264.7 cells were treated with varying concentrations of oxLDL (0.5, 1 and 2 µg/ml) in the presence or absence of SH-GQDs and cells were stained with peroxide-sensitive fluorescent dye (DCFDA). Flow cytometry analysis was performed to investigate the expression of MSR and ATP-binding cassette transporter (ABCA1) after such treatments as the negative control, oxLDL treatment and oxLDL treatment in the presence of either GQDs or SH-GQDs. RESULTS: SH-GQDs had a size ranging from 10 to 30 nm with an average size of 21.3 ± 5.2 nm. The elemental analysis indicated that SH-GQDs are mainly composed of carbon, nitrogen, oxygen and sulfur. The expression levels of ABCA1 in macrophages treated with either LDL or oxLDL were lower than those treated with the media control (the negative control: 100 ± 7.6%; LDL: 82.7 ± 1.2%; and oxLDL: 79.2 ± 1.7%). The level of ABCA1 expression increased as cells were incubated with SH-GQDs (SH-GQDs: 101.5 ± 3.1%). The level of MSR on the surface of macrophages upon being treated with SH-GQDs was lower than those with oxLDL (oxLDL: 112.1 ± 8.8% and SH-GQDs: 91.5 ± 4.2%). CONCLUSION: The enhancement of lipid efflux and down-regulation of MSR in macrophages by SH-GQDs supported its promising usage as a theranostic nanocarrier to prevent foam cell formation and plaque development.

A cascade computer model for mocrobicide diffusivity from mucoadhesive formulations.

BACKGROUND: The cascade computer model (CCM) was designed as a machine-learning feature platform for prediction of drug diffusivity from the mucoadhesive formulations. Three basic models (the statistical regression model, the K nearest neighbor model and the modified version of the back propagation neural network) in CCM operate sequentially in close collaboration with each other, employing the estimated value obtained from the afore-positioned base model as an input value to the next-positioned base model in the cascade. The effects of various parameters on the pharmacological efficacy of a female controlled drug delivery system (FcDDS) intended for prevention of women from HIV-1 infection were evaluated using an in vitro apparatus "Simulant Vaginal System" (SVS). We used computer simulations to explicitly examine the changes in drug diffusivity from FcDDS and determine the prognostic potency of each variable for in vivo prediction of formulation efficacy. The results obtained using the CCM approach were compared with those from individual multiple regression model. RESULTS: CCM significantly lowered the percentage mean error (PME) and enhanced r(2) values as compared with those from the multiple regression models. It was noted that CCM generated the PME value of 21.82 at 48169 epoch iterations, which is significantly improved from the PME value of 29.91% at 118344 epochs by the back propagation network model. The results of this study indicated that the sequential ensemble of the classifiers allowed for an accurate prediction of the domain with significantly lowered variance and considerably reduces the time required for training phase. CONCLUSION: CCM is accurate, easy to operate, time and cost-effective, and thus, can serve as a valuable tool for prediction of drug diffusivity from mucoadhesive formulations. CCM may yield new insights into understanding how drugs are diffused from the carrier systems and exert their efficacies under various clinical conditions.

Development of Man-rGO for Targeted Eradication of Macrophage Ablation.

This study was aimed to develop and evaluate a smart nanosystem that targeted photothermal ablation of inflammatory macrophages in atherosclerotic plaque. Mannosylated-reduced graphene oxide (Man-rGO) was synthesized using three step procedures: (1) preparation of ox-GOs, (2) microwave-assisted synthesis of PEI-rGOs, and (3) mannosylation of PEI-rGO using reductive amination reaction (Man-rGOs). The ζ-potential of Man-rGO that signifies electrophoretic mobility of the charged surface was examined using Zetasizer Nano ZS. The effects of Man-rGO on the cell viability was evaluated using LDH assay and AlamarBlue assay. The targeting efficacy of Man-rGO was assessed using the cellular uptake rate by M2-polarized (i.e., which is induced by IL-4) macrophage. The effects of NOMela loaded in Man-rGO on the enhancement of phagocytosis were evaluated by examining the phagocytic clearance rate of zymosan-FITC particles. The microwave-assisted reduction of GOs was adapted for a facile synthesis of polyethylenimine-reduced GO (PEI-rGO). The mannose functionalization (Man-rGO) of PEI-rGO produced a greater number of amide linkages formed by reductive amination reaction between PEI-rGO and mannose. The ζ-potential of PEI-rGO was +30.6 ± 3.3 mV, whereas that of Man-rGO was down to +13.1 ± 3.8 mV upon interaction with mannose mainly due to the conjugation of mannose on the PEI-rGO surface. Near-infrared (NIR) irradiation increased the temperature of Man-rGO solution to around 45 °C, suggesting that Man-rGO is more potent than ox-GO or rGO in photothermal ablation activity triggered by NIR laser irradiation (808 nm). All testing formulations at the concentrations up to 10 µg/mL exerted less than 10% of membrane disintegration. For AlamarBlue study, more than 90% of cell viability were maintained at the concentrations (up to 10 µg/mL) of all tested formulations. The fluorescent microscopy images of cells after 1 h incubation demonstrated that Man-rGO were mainly accumulated at the subcellular level where the mannose receptors were overexpressed. The cell viability of macrophages significantly decreased upon exposure to Man-rGO irradiated with NIR, but no changes were observed from that of mast cells (for mast cells, 98.3 ± 0.3%; for macrophages, 67.8 ± 1.3%, p < 0.01), indicating that Man-rGO achieved enhanced targetability toward mannose receptor mediated cellular uptake. N-Nitrosomelatonin (NOMela) loaded in macrophage exerted enhanced phagocytic activity. It was concluded that the enhanced photothermal ablation activity of Man-rGO triggered by NIR laser irradiation was mediated through their targetability toward overexpressed mannose receptor, a marker of M2-phenotype of macrophage. The results of this study supported that Man-rGO can serve as an efficient platform for the targeted therapy against atherosclerosis.

Biomimicking Robust Hydrogel for the Mesenchymal Stem Cell Carrier.

PURPOSE: This study was aimed to develop a hydrogel-nanofiber as an advanced carrier for adipose derived human mesenchymal stem cells (AD-MSCs) and evaluate its potential for immunomodulatory therapies applicable to surface coating of drug eluting stent (DES) against coronary artery diseases (CAD). METHODS: A mixture of dispersing-nanofibers (dNFs) and poly (ethylene glycol)-diacrylate (PEGDA) were blended with sodium alginate to achieve robust mechanical strength. The effects of stem cell niche on cell viability and proliferation rates were evaluated using LDH assay and alamar blue assay, respectively. The amount of Nile-red microparticles (NR-MPs) remained in the hydrogel scaffolds was examined as an index for the physical strength of hydrogels. To evaluate the immunomodulatory activity of AD-MSCs as well as their influence by ROS, the level of L-Kynurenine was determined as tryptophan replacement compounds in parallel with IDO secreted from AD-MSCs using a colorimetric assay of L-amino acid. RESULTS: Both SA-cys-PEG and SA-cys-dNF-PEG upon being coated on stents using electrophoretic deposition technique displayed superior mechanical properties against the perfused flow. d-NFs had a significant impact on the stability of SA-cys-dNF-PEG, as evidenced by the substantial amount of NR-MPs remained in them. An enhanced subcellular level of ROS by spheroidal cluster yielded the high concentrations of L-Kynurenine (1.67 ± 0.6 µM without H2O2, 5.2 ± 1.14 µM with 50 µM of H2O2 and 8.8 ± 0.51 µM with 100 µM of H2O2), supporting the IDO-mediated tryptophan replacement process. CONCLUSION: The "mud-and-straw" hydrogels are robust in mechanical property and can serve as an ideal niche for AD-MSCs with immunomodulatory effects.

Nanofiber-coated drug eluting stent for the stabilization of mast cells.

PURPOSE: The nanofiber-hydrogel blend containing nitric oxide (NO) donors and reactive oxygen species (ROS) scavengers (Edaravone: EDV) was explored as an advanced strategy for stabilization of Mast cells (MCs) to achieve efficient immune-suppressive effects. METHODS: Three types of nanofiber hydrogel composites (Bare-Nanofibers (BNF), Nanofiber-Hydrogels (NF-Gel) and Cross-linked Nanofiber-Hydrogels (NF-Gel-X)), were evaluated. The degranulation rates of MCs were determined by measurement of the extracellular levels of hydrogen peroxide and the released amounts of ß-hexosaminidase from the activated-MCs (a-MCs). In addition, the effects of EDV on the selective scavenging of the oxygen radicals and prevention of peroxynitrite formation were evaluated. The roles of a-MCs in re-endothelialization and viability of coronary artery endothelial cells (hPCAECs) were defined using alamar blue and LDH assay, respectively. RESULTS: Each polymer matrix has unique morphological characteristics. The effects of EDV (~1.0 mM) on the production of NO were greatly influenced by the presence of superoxide or hydroxyl radicals. NF-G-X containing a mixture of EDV and S-Nitroglutathione (GSNO) produced the highest level of NO under the oxidative stress conditions. GSNO alone or a mixture of GSNO and EDV significantly lowered the degranulation rate of a-MCs (GSNO only: 55.8 ± 5.4%; GSNO with EDV: 50.6 ± 0.6%), indicating that NO plays an integral role in degranulation of a-MCs. There were no significant biochemical evidences of cytotoxic effects of GSNO and EDV on the hPCAECs. CONCLUSIONS: Nanofibers containing a mixture of nitric oxide donors and ROS scavengers could be used as a promising strategy to stabilize MCs from the ROS-mediated immune responses.

Calcium and phosphorus concentrations in native and decellularized semilunar valve tissues.

BACKGROUND AND AIM OF THE STUDY: Native, allograft, xenograft and bioprosthetic semilunar valves are all susceptible to calcific degeneration. However, intrinsic differences in baseline calcium and phosphorus tissue concentrations within mammalian normal valve structural components (e.g., cusps, sinus, vessel wall) additionally subdivided by tripartite regions (e.g., right-, left- and non-coronary leaflets) have never been systematically measured and reported. It was originally hypothesized that variations in normative tissue concentrations of calcium and phosphorus may correspond to subsequent clinical patterns of acquired dystrophic calcification; decellularization was also expected to reduce the tissue concentrations of these elements. METHODS: Native semilunar valves were freshly harvested from 12 juvenile sheep. Half of the valves were decellularized (six aortic and six pulmonary), while the other valves were flash-frozen at -80 degrees C within minutes of euthanasia as native valves. Elemental calcium and phosphorus concentrations were measured in the great vessels, sinus walls and cusps using inductively coupled plasma optical emission spectrometry (ICP-OES), and analyzed with non-parametric statistical tests. RESULTS: Calcium concentrations (microg/mg tissue; median (range) were similar in aortic native cusps (0.37 (0.21)), sinus walls (0.37 (0.09)) and aorta (0.37 (0.08)) (p = 0.8298). Pulmonary calcium concentrations were similar in cusps, but 10-25% higher in the native sinus (p = 0.0018) and pulmonary artery (p < 0.0001) compared to analogous aortic structures. All cusps had higher phosphorus concentrations than their respective conduit tissues. No tripartite regional variations were observed. Decellularization did not reduce the calcium content of cusps, but removed 50-55% of vessel and sinus wall calcium. However, up to 85% of phosphorus was removed from all valve tissues (p < 0.001). CONCLUSION: There were no significant differences in normal tissue concentrations of calcium between aortic valve functional structures, and no semilunar tripartite regional differences in either semilunar valve complex. Thus, the distribution of baseline tissue calcium content of healthy young valves is not inherently predictive of selective or asymmetric anatomical patterns of valve degenerative calcification. Native semilunar cusps contain the highest phosphorus concentrations. Decellularization reduces all elemental concentrations except for cuspal calcium.

Advanced cardiovascular stent coated with nanofiber.

Nanofiber was explored as a stent surface coating substance for the treatment of coronary artery diseases (CAD). Nanofibers loaded with nanoparticles containing ß-estradiol were developed and exploited to prevent stent-induced restenosis through regulation of the reactive oxygen species (ROS). Eudragit S-100 (ES), a versatile polymer, was used as a nanoparticle (NP) base, and the mixtures of hexafluoro-2-propanol (HFIP), PLGA and PLA at varying ratios were used as a nanofiber base. ß-Estradiol was used as a primary compound to alleviate the ROS activity at the subcellular level. Nile-Red was used as a visual marker. Stent was coated with nanofibers produced by electrospinning technique comprising the two-step process. Eudragit nanoparticles (ES-NP) as well as 4 modified types of NP-W (ES-NP were dispersed in H2O, which was mixed with HFIP (1:1 (v/v) and then subsequently added with 15% PLGA), NP-HW (ES-NP were dispersed in H2O, which was mixed with HFIP (1:1 (v/v)) already containing 15% PLGA), NP-CHA (ES-NP with a chitosan layer were added in H2O, which was mixed with HFIP (1:1 (v/v)) containing 15% PLGA), and NP-CHB (ES-NP with a chitosan layer were added in H2O, which was mixed with HFIP (1:1 (v/v)) containing the mixture of PLGA and PLA at a ratio of 4:1) were developed, and their properties, such as the loading capacity of ß-estradiol, the release profiles of ß-estradiol, cell cytotoxicity and antioxidant responses to ROS, were characterized and compared. Among composite nanofibers loaded with nanoparticles, NP-CHB had the maximal yield and drug-loading amount of 66.5 ± 3.7% and 147.9 ± 10.1 µg, respectively. The nanofibers of NP-CHB coated on metallic mandrel offered the most sustained release profile of ß-estradiol. In the confocal microscopy study, NP-W exhibited a low fluorescent intensity of Nile-Red as compared with NP-HW, indicating that the stability of nanoparticles decreased, as the percentage volume of the organic solvent increased. Nanofibers incorporated with ß-estradiol yielded a high endothelial proliferation rate, which was about 3-fold greater than the control (without ß-estradiol). The cells treated with the enhanced level of H2O2 (>1 mM: as ROS source) were mostly nonviable (81.1 ± 12.4%, p < 0.01), indicating that ROS induce cell apoptosis and trigger the rupture of atheroma thin layer in a concentration dependent manner. Nanofibers containing ß-estradiol (0.5 mM) lowered cellular cytotoxicity from 25.2 ± 4.9% to 8.1 ± 1.4% in the presence of 600 µM H2O2, and from 86.8 ± 8.4% to 59.4 ± 8.7% in the presence of 1.0 mM H2O2, suggesting that ß-estradiol efficiently protected hPCECs from ROS induced cytotoxicity. The level of NO production in hPCECs in the presence of ß-estradiol after 6 days of incubation was much greater than that of the control without ß-estradiol. In summary, nanofibers loaded with nanoparticles containing ß-estradiol could be used as a suitable platform for the surface coating of a cardiovascular stent, achieving enhanced endothelialization at the implanted sites of blood vessels.

Non-alcoholic fatty liver disease and type 2 diabetes: An update.

The global prevalence of non-alcoholic fatty liver disease (NAFLD) is rising, along with the epidemic of diabesity. NAFLD is present in >70% of individuals with type 2 diabetes. Although the mutually detrimental relationship between NAFLD and type 2 diabetes has been well established, a multitude of recent studies have further shown that type 2 diabetes is closely linked to the development of cirrhosis, hepatocellular carcinoma, liver-related morbidity and mortality. In contrast, NAFLD also negatively impacts type 2 diabetes both in terms of its incidence and related adverse clinical outcomes, including cardiovascular and chronic kidney diseases. In response to these global health threats, clinical care pathways for NAFLD and guidelines for metabolic dysfunction-associated fatty liver disease have been developed. Several antidiabetic agents have been evaluated for their potential hepatic benefits with promising results. Furthermore, type 2 diabetes patients are increasingly represented in clinical trials of novel therapeutics for NAFLD. However, despite the wealth of knowledge in NAFLD and type 2 diabetes, lack of awareness of the disease and the potential weight of this problem remains a major challenge, especially among clinicians who are outside the field of hepatology and gastroenterology. This review therefore aimed to provide all diabetes care providers with a summary of the latest evidence that supports NAFLD as an emerging diabetic complication of increasing importance, and to present the current recommendations, focusing on the assessment and therapeutic strategies, on the management of NAFLD among type 2 diabetes patients.

Direct activation of endothelial cells by SARS-CoV-2 nucleocapsid protein is blocked by Simvastatin.

Emerging evidence suggests that endothelial activation plays a central role in the pathogenesis of acute respiratory distress syndrome (ARDS) and multi-organ failure in patients with COVID-19. However, the molecular mechanisms underlying endothelial activation in COVID-19 patients remain unclear. In this study, the SARS-CoV-2 viral proteins that potently activate human endothelial cells were screened to elucidate the molecular mechanisms involved with endothelial activation. It was found that nucleocapsid protein (NP) of SARS-CoV-2 significantly activated human endothelial cells through TLR2/NF-κB and MAPK signaling pathways. Moreover, by screening a natural microbial compound library containing 154 natural compounds, simvastatin was identified as a potent inhibitor of NP-induced endothelial activation. Remarkablely, though the protein sequences of N proteins from coronaviruses are highly conserved, only NP from SARS-CoV-2 induced endothelial activation. The NPs from other coronaviruses such as SARS-CoV, MERS-CoV, HUB1-CoV and influenza virus H1N1 did not affect endothelial activation. These findings are well consistent with the results from clinical investigations showing broad endotheliitis and organ injury in severe COVID-19 patients. In conclusion, the study provides insights on SARS-CoV-2-induced vasculopathy and coagulopathy, and suggests that simvastatin, an FDA-approved lipid-lowering drug, may benefit to prevent the pathogenesis and improve the outcome of COVID-19 patients.

The involvement of intracellular calcium in the MCT-mediated uptake of lactic acid by HeLa cells.

The main object of this study was to evaluate the role of intracellular free calcium ion [Ca2+](in) in monocarboxylate transporter (MCT) mediated drug uptake by HeLa cells. It was hypothesized that alterations in the [Ca2+](in) levels affect Na+-H+ exchanger (NHE) regulated pH(in) and thereby produce the proton-motivated driving force for monocarboxylate mediated substrate transport. The changes in intracellular pH (pH(in)) and MCT mediated uptake rates of L-lactic acid by HeLa cells, a human cervical adenocarcinoma cell line, were evaluated under the conditions, whose [Ca2+](in) concentrations were altered by various calcium modulators, such as EGTA-AM (a chelator), nifedipine (a Ca2+ channel antagonist) and A23187 (an ionophore). For the purpose of comparison, the L-lactic acid uptake by HeLa cells was also evaluated under various pH(in) conditions induced by dexamethasone. The effects of the extracellular sodium concentration on the L-lactic acid uptake by HeLa cells were evaluated to determine the involvement of NHE-regulated pH changes in the MCT mediated drug uptake process. The [Ca2+](in) concentrations and pH(in) in HeLa were assessed using fluorescent probes fura-2 and 2',7'-bis[2-carboxyethyl-5-carboxyfluorescein] (BCECF), respectively. The treatment of HeLa cells with A23187 at concentrations of 50 and 100 microM enhanced [Ca2+](in) by 100% and 200% of the control, respectively. EGTA/AM (50 microM) or nifedipine (100 microM) did not cause any significant changes in the [Ca2+](in) levels, whereas EGTA/AM (100 microM) and nifedipine (200 microM) reduced the [Ca2+](in) levels by 30% and 25%, respectively, as compared with the control. A23187 at a concentration of 100 microM in the incubation medium lowered pH(in) (pH 5) and subsequently the uptake rate of lactic acid by 50% (0.47 +/- 0.03 micromol/mg protein/min) of the control. In contrast, nifedipine (200 microM) and EGTA-AM (100 microM), the calcium modulators that lowered the [Ca2+](in) levels and maintained the higher pH(in) (pH > 6) of HeLa cells, enhanced the uptake rate of lactic acid by 60% and 130% of the control, respectively. The results of this study demonstrated that there was a close correlation between the [Ca2+](in) level and pH(in) and that NHEs were involved with the MCT mediated uptake process in HeLa cells. An understanding of the role of [Ca2+](in) in the MCT mediated transport process could provide an efficient strategy to improve the systemic delivery of monocarboxylate substrates through the cervical mucosa.

Deep Learning in Personalization of Cardiovascular Stents.

Deep learning (DL) application has demonstrated its enormous potential in accomplishing biomedical tasks, such as vessel segmentation, brain visualization, and speech recognition. This review article has mainly covered recent advances in the principles of DL algorithms, existing DL software, and designing strategies of DL models. Latest progresses in cardiovascular devices, especially DL-based cardiovascular stent used for angioplasty, differential and advanced diagnostic means, and the treatment outcomes involved with coronary artery disease (CAD), are discussed. Also presented is DL-based discovery of new materials and future medical technologies that will facilitate the development of tailored and personalized treatment strategies by identifying and forecasting individual impending risks of cardiovascular diseases.