Stankevičius, Edgaras

Extracellular vesicles (EVs) released by glioblastoma (GBM) cells circulate systemically and modulate tumour-host interactions, influencing vascular function both within and beyond the tumour microenvironment. The EVs span a size range from tens to hundreds of nanometres; however, the specific effects of distinct GBM-derived EV size subpopulations on endothelial function across different endothelial cell types remain poorly understood.

The transient receptor potential vanilloid 4 channel (TRPV4) is thought to play a pivotal role in pulmonary arterial circulation. The present study hypothesizes that TRPV4 activation increases nitric oxide (NO) release and activates calcium-activated potassium of intermediate conductance (KCa3.1) in pulmonary arteries. Pulmonary arteries were isolated from wild-type mice (wt) and mice deficient in KCa3.1 channels (Kcnn4−/−) and mounted for simultaneous NO concentration and relaxation measurements. Human small pulmonary arteries were isolated and mounted in microvascular myographs for isometric tension recordings. Acetylcholine-induced increases in NO and relaxation of pulmonary arteries were slightly decreased in pulmonary arteries from Kcnn4−/− versus wt mice. An activator of TRPV4 channels, GSK1016790A, increased NO and relaxation to the same degree in pulmonary arteries from wt and Kcnn4−/− mice. A blocker of TRPV4 channels, HC06704, inhibited increases in NO concentration with no effect on acetylcholine (ACh) relaxation in pulmonary arteries from wt mice, but blocked increases in NO concentration and relaxation in pulmonary arteries from Kcnn4−/− mice and responses to GSK1016790A in pulmonary arteries from wt and Kcnn4−/− mice. Concentration-dependent relaxations induced by an inhibitor of sarcoplasmic Ca-ATPase, cyclopiazonic acid, were blocked in the presence of an inhibitor of NO synthase and a blocker of KCa3.1 channels, TRAM-34, in pulmonary arteries from wt mice, but were unaltered in the presence of TRAM-34 in arteries from Kcnn4−/− mice, or the presence of a blocker of TRPV4 channels. In small human pulmonary arteries, ACh and sodium nitroprusside (SNP) induced concentration-dependent relaxations, blocked by endothelial cell removal, in the presence of an inhibitor of NO synthase and the KCa3.1 channel blocker TRAM-34. GSK1016790A induced relaxation of human pulmonary arteries with endothelium, but failed to relax arteries without endothelium. The findings suggest that TRPV4 channels are involved in endothelium-dependent relaxation and likely regulate pulmonary vascular tone by modulating NO release.

Graphical abstract https://www.sciencedirect.com/science/article/pii/S1773224726001000?via%3Dihub

Environmental exposures are increasingly linked to reproductive dysfunctions such as endometriosis, ovarian insufficiency, and polycystic ovary syndrome (PCOS). Through endocrine disruption, oxidative stress, and epigenetic pathways, heavy metals (such as cadmium [Cd], lead [Pb], mercury [Hg], and arsenic [As]) and trace elements (such as zinc [Zn], copper [Cu], and selenium [Se]) may affect female fertility. Nevertheless, there are still few integrated assessments that address their combined consequences. The goal is to perform a critical evaluation and systematic analysis of epidemiological data about the link between reproductive health issues in women of reproductive age and exposure to heavy metals and trace elements.

Recent biobank studies have identified a link between peripheral viral infections and neurodegeneration, yet the mechanisms underlying this connection remain poorly defined. This study examines whether extracellular vesicles (EVs) released from virally stimulated airway cells contribute to vascular and blood-brain barrier (BBB) dysfunction, thereby bridging a critical gap in our understanding of how peripheral viral infections drive neuroinflammation. Specifically, we evaluate the effects of EVs derived from airway epithelial cells stimulated with either polyinosinic-polycytidylic acid (poly I:C) or the SARS-CoV-2 receptor-binding domain (RBD) on hCMEC/D3, a human cerebral microvascular endothelial cell line that serves as an in vitro model of the BBB. Human bronchial epithelial cells (HBEC3-KT) were cultured in AlphaMEM medium, supplemented with 10% FBS, grown to ~90% confluency, and treated with either poly IC (12€ ŵg/mL, 2 h) or the RBD (100 ng/mL, 48 h), while untreated cells were used as controls EVs were isolated from conditioned cell culture medium supplemented with exosome-depleted serum using the prExo (Exolitus) precipitation-based isolation kit and characterised by nanoparticle tracking analysis (NTA) for size distribution and concentration, and cryogenic transmission electron microscopy (cryoTEM) for vesicle morphology. hCMEC/D3 were seeded and allowed to form confluent monolayers (4 days post-seeding) before treatment with EVs for 48 h. Endothelial cells metabolic activity was assessed using the PrestoBlue cell viability assay, mitochondrial function via Seahorse XF Mito Stress Test, BBB integrity by transendothelial electrical resistance (TEER), and endothelial migration by scratch wound healing assay. Statistical analyses were performed in GraphPad Prism. All EV isolation and characterisation procedures were conducted in accordance with the latest MISEV 2023 guidelines. Isolated EVs, mostly, were 100-150 nm diameter and had lipid bilayer. EVs from poly I:C- and RBD-stimulated airway cells significantly reduced hCMEC/D3 metabolic activity at 5A-10à particles/20000 cells, compared to treatment of EVs from healthy untreated airway cells. Seahorse analysis revealed decreased maximal respiration and spare respiratory capacity, with increased non-mitochondrial oxygen consumption in both treatment groups in endothelial cells. TEER measurements showed that both poly I:C-EVs and RBD-EVs impaired BBB integrity; however, only RBD-EVs significantly reduced endothelial migration in the scratch assay. EVs released from virally stimulated airway epithelial cells impair BBB endothelial metabolism, mitochondrial function, and barrier integrity, with RBD-EVs exerting additional inhibitory effects on endothelial migration. These findings highlight a potential mechanism by which peripheral viral infections may promote neurovascular dysfunction and contribute to neuroinflammation.

Introduction: Glioblastoma (GBM) is an aggressive brain tumour with rapid progression, therapy resistance, and a complex microenvironment. GBM-derived extracellular vesicles (EVs) circulate systemically, exert size-dependent effects, and mediate vascular abnormalities. Given the heart's high vascularisation, EVs may also drive cardiovascular dysfunction. While cardio-oncology focuses on therapy-related effects, the role of GBM EVs remains unclear. This study investigates how the GBM secretome contributes to cardiotoxicity, emphasising EV size.

Materials & Methods: EVs from the human GBM HROG36 cell line were isolated by centrifugation/ultracentrifugation and characterised by NTA and cryo-EM. Uptake by HUVECs and rat cardiomyocytes was confirmed by immunocytochemistry. Functional effects were assessed in HUVECs (migration, tube formation, TEER, tight junctions), cardiomyocytes (contractility), and both cell types using Seahorse XFp MitoStress analysis.

Results: Both EV populations entered HUVECs and cardiomyocytes within 24 h. After 72 h, large EVs impaired HUVEC tube formation but increased motility, while small EVs reduced TEER, altered ZO-1/claudin-5, and shifted metabolism toward glycolysis. Large EVs slowed contraction and relaxation in cardiomyocytes, whereas small EVs enhanced mitochondrial activity and glycolysis.

Conclusion: GBM-derived EVs exert size-dependent effects, with large EVs disrupting vascular structure and cardiac contractility, and small EVs impairing endothelial barrier function and metabolism.

The use of cardioplegia not only achieves cardiac arrest but also minimizes ischemic/reperfusion (I/R) injury, potentially improving short- or long-term outcomes. The aim of this study was to evaluate the impact of different cardioplegic solutions - del Nido, Custodiol HTK and St. Thomas on genes expression and cytokines response in an immature rat heart model of I/R using the Langendorff preparation. Expression of genes which are involved in cell cycle, proliferation, apoptosis resistance and response to hypoxia were determined in cardiac tissue, as well as levels pro/anti-inflammatory cytokines were measured.

This study evaluates the primary hypothesis of red cell mass (RCM) dependent amplitude of homeostatically acceptable limits of fluctuation in plasma dilution by exploring the correlation between RCM-specific equilibrated hematocrit (EQ_Hct) and amplitude of plasma dilution during perioperative mini Volume Loading Test (mVLT).

This prospective observational clinical study investigated discrepancy between noninvasive continuous haemoglobin (SpHb, Masimo Corp., Irvine, CA) measurements with sensors placed on three fingertips during elective cardiac surgery.

Background and Objectives: Previous studies have shown that a major part of adverse drug reactions (ADRs) are preventable, and they contribute to increased morbidity, mortality, and costs. To our knowledge, no study investigating preventable ADRs has been carried out in Lithuania. Therefore, the aim of this study was to characterize ADRs in the intensive care unit (ICU) of a secondary care Lithuanian hospital as well as to identify drug classes and organ systems most commonly implicated in preventable and nonpreventable ADRs. Materials and Methods: This observational prospective study was conducted in an 18-bed ICU of Kaunas Hospital of the Lithuanian University of Health Sciences from 1 September 2021 to 31 August 2023. All ADRs were assessed for causality, severity, and preventability. The Anatomical Therapeutic and Chemical (ATC) system was used to classify drug classes implicated in ADRs. The organ systems affected were analyzed using the Medical Dictionary for Regulatory Activities (MedDRA). Results: A total of 154 patients with a median age of 78.8 years (range, 18–97) were enrolled into this study. There were 255 ADRs identified; preventable ADRs accounted for 87.5%. Among the preventable ADRs, the top three therapeutic subgroups were antithrombotic agents (26.5%), anti-inflammatory and antirheumatic products (22.0%), and blood substitutes and perfusion solutions (20.2%). Meanwhile, among nonpreventable ADRs, antibacterials for systemic use (62.5%) and antithrombotic agents (46.9%) were the two most common therapeutic subgroups. The gastrointestinal as well as the skin and subcutaneous tissues organ systems were more likely to be affected by nonpreventable ADRs (56.3% vs. 17.5%, p ˂ 0.05 and 12.5% vs. 0.4%, p ˂ 0.05, respectively), while the renal and urinary organ systems were more likely to be affected by preventable ADRs (38.1% vs. 6.3%, p ˂ 0.05). Conclusions: Our study showed a very high incidence of preventable ADRs (87.5%). Drugs affecting blood and blood-forming organs were most frequently implicated in these ADRs. This area deserves special attention and strategies need to be implemented to reduce the incidence of preventable ADRs and their impact on the healthcare system. Moreover, it emphasizes the need for future studies at a national level as, to our knowledge, this is the first study addressing the issues of avoidable harm at the ICU of one Lithuanian hospital.