Endothelial Research Articles

Uric Acid-HDL Ratio as Predictor of Major Cardiovascular Events in Very High Cardiovascular Risk Patients Treated with High-Intensity Statin Therapy

Introduction: Uric acid levels are positively correlated with cardiovascular disease. High Density Lipoprotein (HDL) cholesterol plays a role in suppressing blood oxidation reactions and protecting vascular endothelial cells. The combination of these two metabolic parameters, the Uric Acid-HDL Ratio (UHR) makes it a very useful predictor marker for metabolic disorders. Statins are known to be the gold standard for raising HDL cholesterol. Some types of statins are also known to reduce serum uric acid levels by involving pleiotropic actions. The aim of this study was to determine baseline UHR as a predictor of MACE after administration of high-intensity statins.Objectives: The Objective of this study was to determine baseline UHR as a predictor of MACE after administration of high-intensity statins.Methods: This study was conducted at the cardiac center of Dr Zainal Abidin Regional General Hospital. This study is an observational study with a prospective cohort design. The target population was patients classified as patients with very high cardiovascular risk and using consecutive sampling techniques.Results: During the study period, a total of 82 patients with ischemic heart disease with very high cardiovascular risk met the inclusion and exclusion criteria. Of these, 49 patients (59.8%) were identified as having major cardiovascular events (MACE) while the remaining 33 patients (40.2%) did not had MACE. The results of this study are that statin therapy may not significantly affect the uric acid-HDL ratio in the context of major cardiovascular events.Conclusions: The uric acid-HDL ratio had no significant predictive ability for major cardiovascular events in patients with very high cardiovascular risk who were treated with high-intensity statin therapy.

The factor XI/XIa antibody abelacimab combined with enoxaparin inhibits filter clotting in hemodialysis circuits ex vivo.

Drugs targeting factor XI may offer an alternative to heparin for preventing blood clotting in extracorporeal circulation. We investigated the effects of abelacimab, a novel monoclonal antibody targeting factor XI. We collected whole blood samples into two bags (each 240ml, control group: enoxaparin 1.2mg, treatment group: enoxaparin 1.2mg plus abelacimab 5mg) and circulated in a hemodialysis device for up to 3h. We performed whole blood aggregation and thromboelastometry at several time points. Time to filter clotting was the primary endpoint. We included 10 volunteers. Each volunteer's blood was split into two bags (containing enoxaparin +/- abelacimab) and used simultaneously on two hemodialysis devices. The treatment group's time to filter clotting was significantly prolonged (treatment: 180min, IQR 180-180 vs. control: 120min, IQR 97-147, p < 0.001), and the transmembrane pressure was significantly lower at the end of the circuit flow (treatment: 13 mmHg vs. control: 65 mmHg, p = 0.001). Fibrinogen levels and median platelet counts were preserved. Platelet aggregation was better preserved in the treatment group for ristocetin (p = 0.015), thrombin receptor activating peptide (p = 0.015), and arachidonic acid (p = 0.001). Thromboelastometry showed prolonged clotting times in the treatment group at the end of the experiment (INTEM, p < 0.001; HEPTEM, p = 0.001). Abelacimab prolonged the time to filter clotting in this ex vivo model of hemodialysis. This is an aggressive model due to the frequent re-circulation of blood and a lack of endothelial cells. These data provide support for testing abelacimab in patients on hemodialysis.

Enduring effects of acute prenatal ischemia in rat soleus muscle, and protective role of erythropoietin.

Motor disorders are considered to originate mainly from brain lesions. Placental dysfunction or maternal exposure to a persistently hypoxic environment is a major cause of further motor disorders such as cerebral palsy. Our main goal was to determine the long-term effects of mild intrauterine acute ischemic stress on rat soleus myofibres and whether erythropoietin treatment could prevent these changes. Rat embryos were subjected to ischemic stress at embryonic day E17. They then received an intraperitoneal erythropoietin injection at postnatal days 1-5. Soleus muscles were collected at postnatal day 28. Prenatal ischemic stress durably affected muscle structure, as indicated by the greater fiber cross-sectional area (+ 18%) and the greater number of mature vessels (i.e. vessels with mature endothelial cells) per myofibres (+ 43%), and muscle biochemistry, as shown by changes in signaling pathways involved in protein synthesis/degradation balance (-81% for 4EBP1; -58% for AKT) and Hif1α expression levels (+ 95%). Erythropoietin injection in ischemic pups had a weak protective effect: it increased muscle mass (+ 25% with respect to ischemic pups) and partially prevented the increase in muscle degradation pathways and mature vascularization, whereas it exacerbated the decrease in synthesis pathways. Hence, erythropoietin treatment after acute ischemic stress contributes to muscle adaptation to ischemic conditions.

Systems-level reconstruction of kinase phosphosignaling networks regulating endothelial barrier integrity using temporal data.

Phosphosignaling networks control cellular processes. We built kinase-mediated regulatory networks elicited by thrombin stimulation of brain endothelial cells using two computational strategies: Temporal Pathway Synthesizer (TPS), which uses phosphoproteomics data as input, and Temporally REsolved KInase Network Generation (TREKING), which uses kinase inhibitor screens. TPS and TREKING predicted overlapping barrier-regulatory kinases connected with unique network topology. Each strategy effectively describes regulatory signaling networks and is broadly applicable across biological systems.

Extracellular vesicles derived from endothelial progenitor cells modified by Houshiheisan promote angiogenesis and attenuate cerebral ischemic injury via miR-126/PIK3R2.

Angiogenesis following cerebral ischemia is crucial for restoring blood supply to the ischemic region. Extracellular vesicles (EVs) derived from endothelial progenitor cells (EPCs) offer potential therapeutic benefits in the treatment of cerebral ischemia. Houshiheisan (HSHS) has been shown to improve clinical outcomes in ischemic stroke patients, reduce cerebral ischemic damage in rats, and protect endothelial cells. However, the potential effects of HSHS-modified EPC-derived EVs (EVsHSHS) for cerebral ischemia remain unexplored. This study investigated the impact of EVsHSHS on angiogenesis using rats with permanent middle cerebral artery occlusion (pMCAO) and brain microvascular endothelial cells (BMECs) subjected to oxygen-glucose deprivation (OGD). Results demonstrated that EVsHSHS promoted the proliferation, migration, and tube formation of BMECs in vitro. In vivo, high doses of EVsHSHS exhibited better performance than equivalent doses of unmodified EPC-derived EVs in reducing cerebral infarction volume, improving cortical blood perfusion, decreasing neurological deficit scores, and increasing cortical microvessel density at day 7 post-modeling. The pro-angiogenic effects of EVsHSHS following cerebral ischemia were associated with the regulation of miR-126 and the PIK3R2/PI3K/AKT pathway.

Tumor-derived EBV-miR-BART2-5p promotes nasopharyngeal carcinoma metastasis by inducing pre-metastatic endothelial cell pyroptosis.

Extravasation is a key step in tumor metastasis. Epstein‒Barr virus (EBV) plays a crucial role in nasopharyngeal carcinoma (NPC) metastasis. However, the functions and molecular mechanisms of EBV during tumor cell extravasation remains unclear. Here, we showed that the expression of pyroptosis-associated proteins is greater in the endothelial cells of metastatic NPC tissues than in those of nontumor tissues Exosomes derived from NPC cells promoted endothelial cell pyroptosis, vascular permeability, and tumor cell extravasation. Moreover, we found that BART2-5p is abundant in serum exosomes from NPC patients with metastasis and NPC cells, and that it regulates endothelial cell pyroptosis in pre-metastatic organs via MRE11A. Exosomes containing a BART2-5p inhibitor and AAV-MRE11A attenuated endothelial cell pyroptosis and tumor metastasis. Moreover, in the endothelial cells of metastatic tissues from NPC patients, the BART2-5p level was positively associated with pyroptosis-related protein expression. Collectively, our findings suggest that exosomal BART2-5p is involved in pre-metastatic niche formation, identifying secreted BART2-5p as a potential therapeutic target for NPC metastasis. Implications: The finding that secreted BART2-5p is involved in pre-metastatic niche formation may aid the development of potential therapeutic target for NPC metastasis.

Endothelin-1 receptor blockade impairs invasion patterns in engineered 3D high-grade serous ovarian cancer tumouroids.

High-grade serous ovarian cancer (HG-SOC), accounting for 70-80% of ovarian cancer deaths, is characterized by a widespread and rapid metastatic nature, influenced by diverse cell types, cell-cell interactions, and acellular components of the tumour microenvironment (TME). Within this tumour type, autocrine and paracrine activation of the endothelin-1 receptors (ET-1R), expressed in tumour cells and stromal elements, drives metastatic progression. The lack of three-dimensional models that faithfully recapitulate the unique HG-SOC TME has been the bottleneck in performing drug screening for personalized medicine. Herein, we developed HG-SOC tumouroids by engineering a dense central artificial cancer mass (ACM) containing HG-SOC cells, nested within a compressed hydrogel recapitulating the stromal compartment comprising type I collagen, laminin, fibronectin, and stromal cells (fibroblasts and endothelial cells). ET-1-stimulated HG-SOC cells in the tumouroids showed an altered migration pattern and formed cellular aggregates, mimicking micrometastases that invaded the stroma. Compared with control cells, ET-1-stimulated tumouroids showed a higher number of invasive bodies, which were reduced by treatment with the dual ET-1 receptor (ET-1R) antagonist macitentan. In addition, ET-1 increased the size of the invading aggregates compared with control cells. This study establishes an experimental 3D multicellular model eligible for mechanical research, investigating the impact of matrix stiffness and TME interactions, which will aid drug screening to guide therapeutic decisions in HG-SOC patients.

CORRECTION OF ENDOTHELIAL DYSFUNCTION WITH SULFAPORIN IN HYPERCHOLESTEROLEMIC RABBITS

In animals with experimental hypercholesterolemia, depending on the time of survey, we observed activation of smooth muscle cells and macrophages, causing development of inflammation in the intima, as evidenced by high levels of C-reactive protein (CRP), endothelin-1, and homocysteine. The aim of the study was to investigate the molecular mechanisms of endothelial dysfunction in HCH and the possibility of their correction with sulfaporin. Material and methods of the study Experiments were conducted on 46 Chinchilla rabbits with an average weight of 2.5–3.0 kg, kept on a standard diet. The model of experimental HCH in animals was reproduced by orally administering cholesterol (CS) dissolved in sunflower oil in a ratio of 0.2 g per 1 kg of body weight daily for 3 months. The development of hypercholesterolemia was assessed by the increase in the level of totalcholesterol (TC), low-density lipoproteins (LDL) and high-density lipoproteins (HDL), which were determined on a biochemical analyzer. Results of the study. In this case, a progressive increase in the homocysteine level was established: an increase of 1.72 (P &lt; 0.01); 2.33 (P &lt; 0.001) and 2.89 (P &lt; 0.001) times, respectively, for the periods of cholesterol administration of 1, 2 and 3 months. Considering that hyperhomocysteinemia enhances the capture of LDL by endothelial cells, it was of interest to study the relationship between these indicators. The studies showed that with a cholesterol level in LDL of 2.38 ± 0.27 mmol / l, the homocysteine content is 3.46 ± 0.25 pg / ml. With a cholesterol level in LDL of 4.08 ± 0.10; 5.97 ± 0.09 and 6.48 ± 0.11 mmol/l, the homocysteine content increases to 5.96 ± 0.05; 8.07 ± 0.43 and 9.99 ± 0.17 pg/ml, respectively. Conclusions. Hypercholesterolemia is manifested by activation of smooth muscle cells and macrophages, causing the development of inflammation in the intima, which is confirmed by high levels of CRP, endothelin-1, homocysteine.

Human Tendon-on-a-Chip for Modeling the Myofibroblast Microenvironment in Peritendinous Fibrosis.

Understanding the myofibroblast microenvironment is critical to developing therapies for fibrotic diseases. Here the development of a novel human tendon-on-a-chip (hToC) is reported to model this crosstalk in peritendinous adhesions, which currently lacks biological therapies. The hToC facilitates cellular and paracrine interactions between a vascular component, which contains endothelial cells and monocytes, and a tissue hydrogel component that houses tendon cells and macrophages. It is found that the hToC replicates some aspects of in vivo inflammatory and fibrotic phenotypes in preclinical and clinical samples, including activated mTOR signaling, vascular inflammation, tissue contraction induced by myofibroblast activation, inflammatory cytokines secretion, and transendothelial migration of monocytes to the tissue hydrogel. Transcriptional analysis demonstrates significant overlap in enriched pathways in the hToC with human tenolysis samples, including the activation of mTOR signaling. Rapamycin suppresses the vascular inflammation and fibrotic phenotype in the hToC, which provides proof-of-concept of its utility as an in vitro tool for investigating multicellular crosstalk in fibrosis and testing therapeutics to mitigate it.

The Hidden Dangers: E-Cigarettes, Heated Tobacco, and Their Impact on Oxidative Stress and Atherosclerosis—A Systematic Review and Narrative Synthesis of the Evidence

Electronic cigarettes and heated tobacco products have seen significant growth in sales and usage in recent years. Initially promoted as potentially less harmful alternatives to traditional tobacco, recent scientific evidence has raised serious concerns about the risks they pose, particularly in relation to atherosclerosis. While atherosclerosis has long been associated with conventional tobacco smoking, emerging research suggests that electronic cigarettes and heated tobacco may also contribute to the development of this condition and related cardiovascular complications. In a narrative review, we examined the potential effects of heated tobacco products and electronic cigarettes on oxidative stress and atherosclerosis. Several studies have shown that e-cigarettes and heated tobacco increase oxidative stress through the activation of enzymes such as NADPH oxidase. One of the primary effects of these products is their pro-thrombotic and pro-atherosclerotic impact on endothelial cells and platelets, which promotes inflammatory processes within the arteries. Furthermore, the chemicals found in electronic cigarette liquids may exacerbate inflammation and cause endothelial dysfunction. Furthermore, through a systematic review, we analyzed the effects of chronic exposure to electronic and heated tobacco cigarettes on endothelial function, as assessed by brachial flow-mediated dilation (FMD). Although electronic cigarettes and heated tobacco cigarettes are often perceived as safer alternatives to traditional smoking, they could still present risks to cardiovascular health. It is essential to raise public awareness about the potential dangers associated with these products and implement protective measures, particularly for young people.

Increased caveolin 1 by human antigen R exacerbates Porphyromonas gingivali-induced atherosclerosis by modulating oxidative stress and inflammatory responses

Objective: Many different types of infectious oral diseases have been identified clinically, including chronic periodontitis. Porphyromonas gingivalis is the main pathogen causing chronic periodontitis, which is closely related to atherosclerosis (AS) and can promote the expression levels of caveolin 1 (Cav-1) and induced ribonucleic acid (RNA)-binding protein human antigen R (HuR). However, the roles of Cav-1 and its relationship with HuR in P. gingivalis-mediated AS progression remain largely unknown. Here, we aimed to detect the role and molecular mechanisms of Cav-1 in P. gingivalis-mediated AS. Material and Methods: To investigate the role of Cav-1 in P. gingivalis-mediated AS, we infected human umbilical vein endothelial cells (HUVECs) with P. gingivalis at a multiplicity of infection of 100:1 for 6, 12, and 24 h to simulate P. gingivalis-induced AS models in vitro and then transfected them with Cav-1 small interfering RNA to silence Cav-1. Combining molecular biology experimental techniques such as cell counting kit-8 assay, enzyme-linked immunosorbent assay, immunofluorescence staining, flow cytometry, Western blotting, and Oil Red O staining, and apolipoprotein E-deficient AS model mice, the impacts of Cav-1 on cell viability, inflammation, oxidative stress, apoptosis, Cav-1 and intercellular cell adhesion molecule-1 (ICAM-1) levels, and atherosclerotic plaque formation were investigated. Then, the relationship between Cav-1 and HuR was investigated through biotin pull-down and RNA immunoprecipitation assays, reverse transcription quantitative polymerase chain reaction, and Western blot. Results: P. gingivalis can induce Cav-1 expression in a time- and dose-dependent manner (P &lt; 0.05). This effect can inhibit the proliferation of HUVECs (P &lt; 0.05). Cav-1 interference repressed inflammatory response, reactive oxygen species (ROS) and ICAM-1 levels, and apoptosis in the HUVECs (P &lt; 0.05). Cav-1 messenger RNA was stabilized by HuR, which can bind to the 3’ untranslated region of Cav-1. Increase in HuR level reversed the effects of Cav-1 silencing on ROS and ICAM-1 levels and apoptosis in the HUVECs (P &lt; 0.05). In addition, the levels of inflammatory response, oxidative stress, and atherosclerotic plaque formation induced by P. gingivalis in the mouse model were significantly reduced after Cav-1 expression was inhibited (P &lt; 0.05). Conclusion: HuR-activated Cav-1 may promote atherosclerotic plaque formation by modulating inflammatory response and oxidative stress, leading to AS.

Single cell-spatial transcriptomics and bulk multi-omics analysis of heterogeneity and ecosystems in hepatocellular carcinoma.

This study profiled global single cell-spatial-bulk transcriptome landscapes of hepatocellular carcinoma (HCC) ecosystem from six HCC cases and a non-carcinoma liver control donor. We discovered that intratumoral heterogeneity mainly derived from HCC cells diversity and pervaded the genome-transcriptome-proteome-metabolome network. HCC cells are the core driving force of taming tumor-associated macrophages (TAMs) with pro-tumorigenic phenotypes for favor its dominant growth. Remarkably, M1-types TAMs had been characterized by disturbance of metabolism, poor antigen-presentation and immune-killing abilities. Besides, we found simultaneous cirrhotic and HCC lesions in an individual patient shared common origin and displayed parallel clone evolution via driving disparate immune reprograms for better environmental adaptation. Moreover, endothelial cells exhibited phenotypically conserved but executed differential functions in a space-dependent manner. Further, the spatiotemporal traits of rapid recurrence niche genes were identified and validated by immunohistochemistry. Our data unravels the great significance of HCC cells in shaping vibrant tumor ecosystems corresponding to clinical scenarios.

Factors Influencing Corneal Endothelial Cell Density after Descemet Stripping Automated Endothelial Keratoplasty

Purpose: To identify determinants of corneal endothelial cell density at 12 months after Descemet stripping automated endothelial keratoplasty (DSAEK).Methods: This study included patients with bullous keratopathy who underwent DSAEK with internationally sourced donor corneas from March 2021 to March 2022. We assessed various preoperative factors in recipients (age, cause of bullous keratopathy, intraocular pressure, visual acuity, anterior chamber volume, depth and angle, pupil size, white-to-white corneal diameter, and preoperative corneal endothelial cell density) and donors (age, death to preservation time, preservation to transplantation time, death to transplantation time, corneal endothelial cell density, trephine size, and precut tissue thickness). We analyzed factors influencing corneal endothelial cell density up to 12 months after keratoplasty.Results: This study involved 53 patients, with an average recipient age of 65.9 ± 14.6 years prior to surgery. Donor corneas had an average endothelial cell density of 2,871.6 ± 271.6 cells/mm&lt;sup&gt;2&lt;/sup&gt;. At 6 and 12 months postoperatively, endothelial cell densities had decreased by 36.9% and 47.2%, respectively, compared with preoperative levels. Multivariate linear regression analysis indicated that endothelial cell densities at 3 and 12 months postoperatively were linked to recipient age and anterior chamber depth.Conclusions: Corneal endothelial cell density after DSAEK typically stabilized after initial decline; higher densities were found in older recipients and individuals with shallower anterior chambers.

Effects of Cumulative Dissipated Energy on Corneal Endothelial Cells in Phacoemulsification Based on Nucleus Sclerosis

Purpose: To investigate changes in corneal endothelial cell (CEC) by cumulative dissipated energy (CDE) during phacoemulsification.Methods: Based on the degree of nucleus sclerosis (NS), changes in CECs were compared preoperatively and at 1 and 3 months postoperatively in 67 eyes that underwent phacoemulsification.Results: The mean CDE used during surgery was 4.30 ± 2.31. A comparison of the surgical measurements before and 1 month after surgery revealed significant differences in the cell density (CD) and coefficient of variation (CV) (&lt;i&gt;p&lt;/i&gt; &lt; 0.001, 0.011, respectively). The CD showed significant differences among NS grades 2–6 (&lt;i&gt;p&lt;/i&gt; &lt; 0.001). The CDE increased significantly with higher NS grades (r = 0.809, &lt;i&gt;p&lt;/i&gt; &lt; 0.001). Within the same NS grade, there was a positive correlation between higher CDE and greater CEC loss (r = 0.559, &lt;i&gt;p&lt;/i&gt; = 0.001). CD changes were significantly associated with increasing CDE (r = 0.612, &lt;i&gt;p&lt;/i&gt; &lt; 0.001). The CD loss also increased significantly from NS2 to NS6 (&lt;i&gt;p&lt;/i&gt; &lt; 0.001). At 3 months postoperatively, surgical measurements revealed a significant decrease in CD with increasing NS grade (&lt;i&gt;p&lt;/i&gt; = 0.010).Conclusions: CDE increased with higher NS grades and there was a positive correlation between CDE and CEC loss. Therefore, surgeons should plan surgical techniques preoperatively to minimize CDE as NS increases. Additionally, it is important to assess CEC damage postoperatively based on surgical measurements in patients with high intraoperative CDE.

Possible Role of Endothelial-Derived Cellular and Exosomal-miRNAs in Lipid-Mediated Diabetic Retinopathy: Microarray Studies

Diabetic retinopathy (DR) is a salient cause of blindness worldwide. There is still an immense need to understand the pathophysiology of DR to discover better diagnostic and therapeutic modalities. Human retinal endothelial cells (HRECs) were treated with 15-HETE or D-glucose, then miRNAs were isolated, and a microarray was performed. MirWALK 2 and Ingenuity Pathway Analysis (IPA) were used to analyze the microarray results. Exosomal miRNAs from 15-HETE-treated HRECs were isolated, microarrayed, and then imported into IPA for further analysis. The microarray results showed that 15-HETE downregulated 343 miRNAs and upregulated 297 miRNAs in HRECs. High glucose treatment induced a differential expression of HREC-miRNAs where 185 miRNAs were downregulated and 244 were upregulated. Comparing the impact of 15-HETE versus DG or diabetic mouse retina elaborated commonly changing miRNAs. Pathway and target analysis for miRNAs changed in 15-HETE-treated HRECs revealed multiple targets and pathways that may be involved in 15-HETE-induced retinal endothelial dysfunction. The HREC-exosomal miRNAs were differentially expressed after 15-HETE treatment, with 34 miRNAs downregulated and 45 miRNAs upregulated, impacting different cellular pathways. Here, we show that 15-HETE induces various changes in the cellular and exosomal miRNA profile of HRECs, highlighting the importance of targeting the 12/15 lipoxygenase pathway in DR.

Characterization of exosome-mediated propagation of systemic inflammatory responses into the central nervous system.

The mechanisms through which systemic inflammation exerts its effect on the central nervous system (CNS) are still not completely understood. Exosomes are small (30 to 100nm) membrane-bound extracellular vesicles released by most of the mammalian cells. Exosomes play a vital role in cell-to-cell communication. This includes regulation of inflammatory responses by shuttling mRNAs, miRNAs, and cytokines, both locally and systemically to the neighboring as well as distant cells to further modulate the transcriptional and/or translational states and affect the functional phenotype of those cells that have taken up these exosomes. The role of circulating blood exosomes leading to neuroinflammation during systemic inflammatory conditions was hereby characterized. Serum-derived exosomes from LPS-challenged mice (SDEL) were freshly isolated from the sera of the mice that were earlier treated with LPS and used to study the effects on neuroinflammation. Exosomes isolated from the sera of the mice injected with saline were used as a control. In-vitro studies showed that the SDEL upregulate pro-inflammatory cytokine gene expression in the murine cell lines of microglia (BV-2), astrocytes (C8-D1A), and cerebral microvascular endothelial cells (bEnd.3). To further study their effects in-vivo, SDEL were intravenously injected into normal adult mice. Elevated mRNA expression of pro-inflammatory cytokines was observed in the brains of SDEL recipient mice. Proteomic analysis of the SDEL confirmed the increased expression of inflammatory cytokines in them. Together, these results demonstrate and strengthen the novel role of peripheral circulating exosomes in causing neuroinflammation during systemic inflammatory conditions.

Blood CD45+/CD3+ lymphocyte-released extracellular vesicles and mortality in hospitalized patients with coronavirus disease 2019.

The global pandemic of coronavirus disease 2019 (COVID-19) represented a major public health concern. Growing evidence shows that plasma of COVID-19 patients contains large numbers of circulating extracellular vesicles (cEVs) that correlate with disease severity and recovery. In this study, we sought to characterize the longitudinal cEV signature in critically ill COVID-19 patients during hospitalization and its relation to mortality risk. cEVs were quantitatively and phenotypically analysed in hospitalized non-surviving COVID-19 patients at baseline (n = 42) and before exitus (n = 40) and in 40 healthy volunteers as a reference group by high sensitivity nano flow cytometry using specific markers for parental cell sources and activation. Levels of cEV subtypes differed between patients with severe COVID-19 and healthy subjects, specifically those from platelets and endothelial, inflammatory and viral infected cells, which associate to high mortality risk. In the longitudinal analysis from baseline to the time point immediately preceding death, no changes were found for platelet, pan-leukocyte, and lung epithelial cell-shed cEVs, while endothelial cell releases of EVs (eEVs) significantly differed. Vascular endothelial growth factor receptor 2-positive eEVs were significantly increased before death compared to admission whereas endoglin and E-selectin-containing eEVs did not change. Conversely, lymphocyte (ℓEV), monocyte, macrophage, pericyte and progenitor cell-derived cEVs displayed significant reductions before exitus. Noteworthy, levels of CD45+/CD3+-ℓEVs were significantly associated to the patient's survival time. An evolving cEV profile able to discriminate prompt risk of death during hospitalization has been defined suggesting a role for circulating and vascular cell-derived EVs in COVID-19 pathogenesis.

Research on the TSPAN6 regulating the secretion of ADSCs-Exos through syntenin-1 and promoting wound healing.

Exosomes (Exos) from adipose-derived stem cells (ADSCs) have a high inclusion content and low immunogenicity, which helps to control inflammation and accelerate the healing of wounds. Unfortunately, the yield of exosomes is poor, which raises the expense and lengthens the treatment period in addition to impairing exosomes' therapeutic impact. Thus, one of the key problems that needs to be resolved in the current exosome study is increasing the exosome yield. Tetraspanin-6 (TSPAN6) overexpression and knockdown models of ADSCs were constructed to determine the number of exosomes secreted by each group of cells as well as the number of multivesicular bodies (MVBs) and intraluminal vesicles (ILVs) within the cells. Subsequently, the binding region of the interaction between TSPAN6 and syntenin-1 was identified using the yeast two-hybrid assay, and the interaction itself was identified by immunoprecipitation. Finally, cellular and animal studies were conducted to investigate the role of each class of exosomes. When compared to the control group, the number of intracellular MVBs and ILVs was significantly larger, and the number of ADSCsTSPAN6+-Exos was more than three times higher. However, TSPAN6's ability to stimulate exosome secretion was reduced as a result of syntenin-1 knockdown. Additional yeast two-hybrid assay demonstrated that the critical structures for their interaction were the N-terminal, Postsynaptic density protein 95/Discs large protein/Zonula occludens 1 (PDZ1), and PDZ2 domains of syntenin-1, and the C-terminal of TSPAN6. In animal trials, the wound healing rate was best in the ADSCsTSPAN6+-Exos group, while cellular experiments demonstrated that ADSCsTSPAN6+-Exos better enhanced the proliferation and migration of human skin fibroblasts (HSFs) and human umbilical vein endothelial cells (HUVECs). TSPAN6 stimulates exosome secretion and formation, as well as the creation of MVBs and ILVs in ADSCs. Syntenin-1 is essential for TSPAN6's stimulation of ADSCs-Exos secretion. Furthermore, ADSCsTSPAN6+-Exos has a greater ability to support wound healing, angiogenesis, and the proliferation and migration of a variety of cells.

Large Scale Ultrafast Manufacturing of Wireless Soft Bioelectronics Enabled by Autonomous Robot Arm Printing Assisted by a Computer Vision-Enabled Guidance System for Personalized Wound Healing.

A Customized wound patch for Advanced tissue Regeneration with Electric field (CARE), featuring an autonomous robot arm printing system guided by a computer vision-enabled guidance system for fast image recognition is introduced. CARE addresses the growing demand for flexible, stretchable, and wireless adhesive bioelectronics tailored for electrotherapy, which is suitable for rapid adaptation to individual patients and practical implementation in a comfortable design. The visual guidance system integrating a 6-axis robot arm enables scans from multiple angles to provide a 3D map of complex and curved wounds. The size of electrodes and the geometries of power-receiving coil are essential components of the CARE and are determined by a MATLAB simulation, ensuring efficient wireless power transfer. Three heterogeneous inks possessing different rheological behaviors can be extruded and printed sequentially on the flexible substrates, supporting fast manufacturing of large customized bioelectronic patches. CARE can stimulate wounds up to 10 mm in depth with an electric field strength of 88.8 mVmm-1. In vitro studies reveal the ability to accelerate cell migration by a factor of 1.6 and 1.9 for human dermal fibroblasts and human umbilical vein endothelial cells, respectively. This study highlights the potential of CARE as a clinical wound therapy method to accelerate healing.

Vascular hamartoma in horse limbs: presentation, diagnosis, and treatment of two case studies.

Vascular hamartoma is an abnormal development of endothelial tissue that rarely affects horses. Histopathological examination remains the gold standard for definitive diagnosis, yet the presence of abnormal vascular structures must be carefully differentiated from other vascular anomalies and neoplasms. This report describes two cases of vascular proliferation affecting the skin on the lateral side over the left tarsus and the dorsolateral region over the fetlock joint of two fillies. The clinical history, presentation, and radiographic and sonographic findings in both patients suggested a neoplasm of unknown origin. Surgical excision of the mass was successfully performed in both patients. In one patient, a second surgery was performed six months after the first surgery because of the recurrence of the mass. Histopathological examination revealed a disorganized proliferation of well-differentiated, tortuous, and varied-sized blood vessels hemmed in myxoid tissue with abundant fibrocollagenous bands. Immunohistochemical examination of the endothelium markers factor VIII and ERG revealed positive staining of the mass samples in both cases, confirming their vascular origin. The combination of these markers was effective, providing a reliable option for differentiating vascular proliferation in horses. To the best of the author's knowledge, this is the first report applying the ERG endothelial marker to horse vascular malformations.